Neurology of latent nystagmus.

We report eye movement findings in 30 patients with latent nystagmus and who were found to have a variety of associated oculomotor disorders. Latent nystagmus is defined clinically as nystagmus which appears on covering one eye and beats towards the uncovered eye. Recordings showed that the latent nystagmus in 28 patients had slow phases with linear or exponentially decreasing velocity. This nystagmus is termed 'LN'. In 13 of these patients certain manoeuvres (e.g. pursuit) provoked nystagmus with exponentially increasing slow phase velocities characteristic of the congenital form of nystagmus termed 'CN' and we propose that this is a forme fruste of CN. In two patients the nystagmus provoked by cover was latent CN. Twenty-nine patients had a history of strabismus and one had a marked phoria. Some patients had amblyopia whilst others had normal vision in each eye. Although binocular vision was usually absent, six patients had varying degrees of stereopsis. A temporonasal predominance of monocularly elicited optokinetic response previously associated with LN, was present only in a minority of patients. Some responses were bidirectionally absent or of low velocity, possibly the result of a cortical impairment of visual motion detection. The most deranged responses had slow phases which were in the opposite direction to the stimulus as described in CN. The presence of 'forme fruste' CN in many of these patients suggests that some of the derangements of optokinetic responses are due to CN. The findings indicate a greater overlap between the incidences of LN and CN than previously estimated. Thirty percent of patients had large saccadic 'square wave' intrusions. These were not present when there was marked amblyopia. They are attributed to a competitive incongruence of visual fields and eye positions. Dissociations found between the presence and severity of strabismus, stereopsis, amblyopia and optokinetic abnormalities point to these features being relatively independent although associated in typical clusterings. This is evidence against the theory that strabismus and LN are directly caused by nasotemporal optokinetic imbalance which persists because of failure to develop binocular vision. The variability of findings favours the view that LN and CN arise from a genetic or acquired embryological disorder with various degrees and directions of expression.


Comparison of predictable smooth ocular and combined eye-head tracking behaviour in patients with lesions affecting the brainstem and cerebellum.

We compared the ability of eight normal subjects and 15 patients with brainstem or cerebellar disease to follow a moving visual stimulus smoothly with either the eyes alone or with combined eye-head tracking. The visual stimulus was either a laser spot (horizontal and vertical planes) or a large rotating disc (torsional plane), which moved at one sinusoidal frequency for each subject. The visually enhanced vestibulo-ocular reflex (VOR) was also measured in each plane. In the horizontal and vertical planes, we found that if tracking gain (gaze velocity/target velocity) for smooth pursuit was close to 1, the gain of combined eye-hand tracking was similar. If the tracking gain during smooth pursuit was less than about 0.7, combined eye-head tracking was usually superior. Most patients, irrespective of diagnosis, showed combined eye-head tracking that was superior to smooth pursuit; only two patients showed the converse. In the torsional plane, in which optokinetic responses were weak, combined eye-head tracking was much superior, and this was the case in both subjects and patients. We found that a linear model, in which an internal ocular tracking signal cancelled the VOR, could account for our findings in most normal subjects in the horizontal and vertical planes, but not in the torsional plane. The model failed to account for tracking behaviour in most patients in any plane, and suggested that the brain may use additional mechanisms to reduce the internal gain of the VOR during combined eye-head tracking. Our results confirm that certain patients who show impairment of smooth-pursuit eye movements preserve their ability to smoothly track a moving target with combined eye-head tracking.


Oestradiol-17 beta in the milk of cows from 6 days before to 14 days after their insemination.

In artificially inseminated cows (AI on day 1) peak concentrations of oestradiol-17 beta in defatted milk occurred at median times of day 0 during the pre-ovulatory period, days -5 to 2, and day 6 during the post-ovulatory period, days 2-15. Median peak concentrations during these periods were approximately 5 pg/ml and 3 pg/ml respectively. There were no significant differences in the timing or magnitude of oestradiol-17 beta concentrations between cows that became pregnant to the AI and those that entered normal length oestrus cycles immediately after AI.


A quantitative study of eye and head movements during smooth pursuit in patients with cerebellar disease.

Eye and head movements were analysed during smooth pursuit in 16 patients with various forms of cerebellar disease. Smooth pursuit gain was reduced across all frequencies and velocities of target motion for the patient group as a whole, during both sinusoidal and pseudo-random target motion. The graded breakdown in the pursuit response, as pseudo-random target motion became less predictable, was of a similar magnitude in patients and controls, implying that the predictive pursuit mechanisms were intact in these patients. During head-free pursuit, when vestibulo-ocular reflex (VOR) suppression was necessary, performance was not significantly different from that observed during head-fixed pursuit in the patient group. This finding is similar to that noted in control subjects, and is consistent with the observation that the VOR gains associated with head movements in darkness were similar in the patient and control groups. The deficits in pursuit and VOR suppression in patients with cerebellar disease therefore represent a decrease in gain in the closed-loop visual feedback pathways with apparent sparing of the predictive pathways.


Saccadic reaction times in patients with frontal and parietal lesions.

The effect of unilateral circumscribed lesions in different areas of the frontal and parietal cortex on the distributions of saccadic reaction times (SRTs) was investigated in 32 patients under four stimulus conditions: (i) gap-random: a target light appeared 200 ms after extinction of a central fixation light randomly at 8 degrees either left or right; (ii) overlap-random: like (i), but the fixation light remained on ('overlap'); (iii) gap-simultaneous: two peripheral lights appeared simultaneously left and right 200 ms after the extinction of the fixation light, one predesignated by instruction as saccadic target; (iv) overlap-simultaneous: like (iii), but the fixation light remained on. Depending on SRT and condition and based on the data of control subjects, we evaluated the percentages of anticipatory (SRT: 0-90 ms), express (91-157 ms) and regular (158-400 ms) saccades, of time (> 400 ms) and direction errors as well as the mean latencies and the standard deviations. These data were compared across control subjects, frontal lobe patients with a lesion in the region of the frontal eye field (FEF), and frontal patients with a lesion outside the FEF, as well as across control subjects, parietal lobe patients with a lesion in the dorsolateral region, and parietal patients with a lesion outside the dorsolateral region. (i) Frontal patients with a lesion in the FEF region showed an increased percentage of express saccades (gap-random condition), especially with saccades directed towards the side of the lesion (ipsilateral). If fixation was not interrupted prior to target appearance (overlap-random), express saccades were largely suppressed, similar to normals. All patients with a frontal lesion, whether in the region of the FEF or not, showed a decreased percentage of contralateral anticipatory saccades (gap-simultaneous), whereas the percentages of direction and time errors were in the normal range, even if more than one stimulus was presented (e.g. overlap-simultaneous condition). (ii) Patients with dorsolateral parietal lesions showed decomposed SRT patterns (high SRT variability, increased percentages of time and direction errors, decreased percentages of express and anticipatory saccades). The decomposition had a contralateral preponderance and increased if more than one stimulus was visible beside the saccadic target. Our results suggest that FEF lesions impair mainly the volitional control over visually triggered express or reflex-like saccades if fixation is disrupted, whereas dorsolateral parietal lesions impair mainly the selection of the visual targets.


Saccadic dysmetria in a patient with a right frontoparietal lesion. The importance of corollary discharge for accurate spatial behaviour.

Double-step experiments have demonstrated that retinotopic coding is inadequate to explain the spatial performance of the saccadic system. In such experiments a subject is asked to make two successive saccades to fixate two sequentially flashed targets each of which disappears before the first saccade. Despite the dissonance thus created between the retinal location of the second target and the saccade necessary to acquire it, normal humans and monkeys perform the task perfectly well. Single unit recording in monkeys indicates that neurons in the superior colliculus, frontal eye fields and in parietal cortex generate a spatially accurate signal during the performance of double-step saccades, which is thought to be obtained by combining a retinotopic signal with a signal corollary to the previous saccadic eye movement. We studied saccadic eye movements in a patient with a right fronto-parietal lesion using single- and double-step tasks. Single saccades into the left (contralesional) hemifield had longer latency and were hypometric relative to those into the right (ipsilesional) hemifield. Varying the initial orbital position had no effect on the latency and accuracy of saccades to left and right retinal stimuli. When the patient was asked to do a double-step task with targets flashed first into the right field and then into the left field, she performed well. When she was asked to do the same task with a target flashed first into the left field and then into the right field she made the first saccade correctly but never acquired the second target, even though this required her to make a saccade in the normal direction to a stimulus that appeared in the normal field. Such a deficit therefore cannot be one of retinotopic or spatial coding, nor can it be one of generating a certain direction of saccade. We suggest that the deficit is a failure of corollary discharge, the inability to register the amplitude and direction of a saccade into the contralesional field, and use that information to update the representation of the location of the next saccade target.


Acute optic neuritis. A cognitive and magnetic resonance imaging study.

Forty-two patients with acute optic neuritis (ON) (mean duration of symptoms 14.5 d) were compared with a matched, normal control group on a battery of tests of attention and information processing speed. Approximately half the sample (55%) had brain abnormalities shown by magnetic resonance imaging (MRI) and were more impaired across a variety of tests compared with those patients without brain lesions or normal controls. There was no difference in psychometric performance other than the pegboard task between the normal control group and those ON patients without brain involvement. Significant correlations were found between total lesion area in the brain and some tests of attention. Results from the Symbol Digit-Substitution Test were particularly sensitive in this regard and could correctly identify 70% of the sample with brain lesions.


The mechanism of facial sweating and cutaneous vascular responses to painful stimulation of the eye.

The vascular response in the forehead and cheeks to irritating the eye with soapy water was measured in 15 normal subjects. Electrodermal activity, which reflects sweating, was also measured from both sides of the forehead. The mechanism of the response was studied in 15 patients with a unilateral lesion of the facial nerve blocking parasympathetic outflow. Pulse amplitude usually increased briefly on both sides of the forehead after the soap was placed in the eye; the response persisted for several minutes on the ipsilateral side after the soap had been washed from the eye. A facial nerve lesion blocked the vascular response on the lesioned side to stimulation of either eye. No consistent change in pulse amplitude was recorded from the cheeks, although a response was observed in a few subjects. Electrodermal responses to ocular irritation were generally larger on the ipsilateral than contralateral side of the forehead; in patients with facial palsy, electrodermal responses were greater on the normally innervated side than on the lesioned side. The findings suggest that irritating the eye induces a trigeminal-parasympathetic vasodilator reflex and local sweating. The restricted distribution of the response indicates that separate parasympathetic vasodilator reflexes might operate for each division of the trigeminal nerve.


Pathological sweating and flushing accompanying the trigeminal lacrimal reflex in patients with cluster headache and in patients with a confirmed site of cervical sympathetic deficit. Evidence for parasympathetic cross-innervation.

Electrodermal responses (as a measure of sweating) and vascular responses to irritation of the eye were investigated in 11 cluster headache patients and, for comparison, in another 24 patients with a confirmed site of lesion in the cervical sympathetic pathway. Seven of the cluster headache patients had ocular and thermoregulatory signs of a postganglionic sympathetic lesion. In these patients, and in six of seven patients with a postganglionic sympathetic lesion from some other cause, the electrodermal response to ocular stimulation was far greater on the denervated side of the forehead than on the sympathetically intact side. This pathological response can be explained by lacrimotor fibres branching into vacant sympathetic sudomotor pathways. The response could account for excessive forehead sweating during attacks of cluster headache, in spite of the presence of Horner's syndrome, because parasympathetic outflow to the lacrimal glands increases during attacks. In patients with diminished sympathetic vasomotor outflow, the vascular response to ocular irritation was also greater on the denervated side of the forehead than on the sympathetically intact side, irrespective of the site of the lesion. The excessive vascular response in sympathetically denervated skin could be caused by adaptive supersensitivity to peptides such as vasoactive intestinal polypeptide, which is known to be released from parasympathetic terminals. The same mechanism might augment vasodilatation during attacks of cluster headache.


Processing of spatial contrast in peripheral vision in Parkinson's disease.

Two experiments were carried out to test the hypothesis, based on anatomical evidence, that contrast gain might be reduced in the retinal periphery in Parkinson's disease. In the first experiment, subjects set contrast thresholds before and after adaptation to a vertical grating of 2 cycles per degree (c/deg), either stationary or oscillating sideways through its spatial period at 8 Hz, presented either in central vision or 7 degrees peripherally. Threshold elevations were similar for central viewing in both patients and controls. However, for peripheral viewing, elevations were greater in the controls, but smaller in the patients, than for central viewing. In the second experiment, a staircase procedure was used to find the contrast of a peripherally viewed grating of either 4 or 1.2 c/deg which apparently matched that of a centrally viewed grating of the same spatial frequency. Patients needed more contrast (about 1.6 times, at both spatial frequencies) than controls for a match. These results suggest that contrast gain may be lowered in the peripheral retina in Parkinson's disease, perhaps because of an abnormality of dopamine amacrine cells, whose density peaks in the peripheral retina.


Sequential arm movements in patients with Parkinson's disease, Huntington's disease and dystonia.

We studied the performance of sequential arm movements in 14 patients with Parkinson's disease, nine patients with Huntington's disease and seven patients with arm dystonia. The results were compared with those from normal subjects. Subjects had to perform each movement of the sequence as fast as possible, stopping as briefly as possible between two successive movements. In one set of experiments, patients with Parkinson's disease drew four different geometrical patterns in a counter-clockwise direction. The patterns consisted of two, three, four and five segments of identical length. In a second set, the subjects drew a pentagon in a counter-clockwise and a clockwise direction and each side of the pentagon singly in a counter-clockwise direction. All three groups of patients were slow in executing movements and in switching from one movement to the next. Only patients with Parkinson's disease took longer to perform the segments at the end of a sequence. In other words, their movement times lengthened progressively as the sequence progressed. This phenomenon could still be recognized when the direction and position of the segments were changed (pentagon drawn in the counter-clockwise and the clockwise direction) and when the extra-time needed, mainly due to the sequential nature of the task, was considered by computing the differences between movement times obtained during drawing of the pentagon and those obtained when each segment was traced singly. This study demonstrates that sequential movements are abnormal in Parkinson's disease, Huntington's disease and dystonia and that in the performance of long motor sequences, the deficit in sequencing movements is exacerbated only in patients with Parkinson's disease.


Anodal and cathodal stimulation of the upper-limb area of the human motor cortex.

In 18 neurologically normal subjects the corticofugal volleys evoked by anodal and cathodal electrical stimulation of the motor cortex were recorded using epidural electrodes at the high-thoracic and low-thoracic regions of the spinal cord during surgery for scoliosis. At and just above threshold, anodal and cathodal stimulation of the upper-limb area and motor cortex produced a D wave that propagated to the low-thoracic region. The stimulus intensity required to produce D waves was significantly lower with anodal stimulation. I waves were recorded at higher stimulus intensities than the D wave but not more readily with cathodal stimulation. There was no significant difference in the extent to which stimulus intensity had to be increased above D-wave threshold to produce I waves with the two stimulus polarities, and the number of I waves was the same when the stimulus was increased by the same amount above D-wave threshold. After withdrawal of isoflurane, I waves could not be recorded when the stimulus intensity was below D-wave threshold with either stimulus polarity. Anodal stimulation over the upper-limb area remained more effective than cathodal stimulation in producing both D and I waves. These results indicate that, at threshold, regardless of anaesthesia, anodal and cathodal stimuli access upper-limb corticospinal neurons directly at a similar site, the anodal stimuli being more effective. In addition, the results suggest that some corticospinal neurons in the upper-limb area of motor cortex have projections to lumbar segments.


Seizures involving secondary sensory and related areas.

Five patients with seizures involving the secondary sensory and/or related areas (SSRA) are presented. Four of five experienced ictal numbness and/or tingling bilaterally and/or axially; this involved fingertips (three patients), lips (two), tongue (two), and was diffuse in one. The fifth patient experienced bilateral ictal pain. Associated ictal symptoms implicating adjacent regions appeared in all five patients, including contralateral clonic movements (two patients), hypersalivation (two), taste (one), vocalization (two), dysphagia (one), and contralateral sensory march (one). Two patients had ictal symptoms suggestive of adjacent temporal lobe involvement. By history, the SSRA was involved at seizure onset in four and by spread in one. All five patients had electroencephalogram (EEG) or subdural EEG supportive evidence of SSRA involvement: ictal (three) and interictal (three). Three patients had lesions in this area shown by magnetic resonance imaging or computerized tomography and all three had histologically proven glial tumours. Relevant experimental physiological and anatomical data are reviewed.


Morphometry of the Sylvian fissure and the corpus callosum, with emphasis on sex differences.

The relationship between anatomical asymmetries in the perisylvian region and the sizes of different regions of the corpus callosum was investigated post-mortem in 40 brains of right-handed hospital admissions (20 males, 20 females) with no cortical involvement. There were no sex differences either in anatomical asymmetries or in regional size of the callosum. There was a negative correlation between the absolute value of Sylvian fissure (planum temporale) asymmetries and the size of the isthmus in males but not in females. Further, there was a significant negative correlation between the size of the Sylvian fissure (or planum temporale) and the size of the callosal mid-body in males but not in females. There was no correlation between the asymmetry of the planum temporale magnitude of left-right and total size of the planum (left+right). These findings constrain theories about the ontogenesis of hemispheric specialization through changes in callosal connectivity and about sex differences in interhemispheric organization.


Changes in brain functional connectivity in Alzheimer-type and multi-infarct dementia.

Clinical and neuropathological evaluation of elderly subjects with dementia has traditionally concentrated upon the focal distribution of brain disease, ignoring changes in the complex connections that link brain areas and that are crucial for cognition. We examined subjects with the two most common forms of dementia in the elderly (dementia of the Alzheimer type or DAT, and multi-infarct dementia or MID); and used electroencephalographic (EEG) coherence to examine the effects of these illnesses on the functional connections between brain areas. We studied coherence between brain areas known to be linked by two different types of connections: (i) dense narrow bands of long corticocortical fibres; (ii) broad complex networks of corticocortical and corticosubcortical fibres. Areas that were linked by dense narrow bands of long corticocortical fibres showed greatly diminished coherence in subjects with DAT; among MID subjects, this coherence was not significantly affected. Areas that were linked by broad connective networks showed the largest decreases in coherence among MID subjects. These findings are consistent with neuropathological evidence that Alzheimer's disease is a neocortical 'disconnection syndrome' in which there is a loss of structural and functional integrity of long corticocortical tracts. The findings further suggest that the vascular disease of MID most prominently affects broad fibre networks that may be more vulnerable to diffuse subcortical vascular damage. A ratio of coherence from complex corticocortical-corticosubcortical networks divided by coherence from long corticocortical tracts correctly classified 76% of subjects into DAT and MID categories. Overall, these results indicate that EEG coherence detects basic pathophysiological differences between subjects with DAT and MID, and that these differences may be clinically useful.


A critical appraisal of neuropsychological correlates of Japanese ideogram (kanji) and phonogram (kana) reading.

Owing to the Japanese language's unique writing system, which consists of phonograms and ideograms, reading impairments of Japanese brain-damaged patients have attracted the interest of many researchers. Past case reports as well as some widely accepted handbooks and textbooks have concluded that a specific aphasia type or lesion site is associated with a particular impairment pattern of phonograms and ideograms in reading. However, the methodology and analytical procedures in previous studies were inadequate for reliable generalizations to be made. First, the test materials were unspecified or inappropriate, or the number of test items was small. Secondly, the conclusions were presented without providing individual performance data to support them. Thirdly, in associating patterns of reading impairment with lesion sites, only single cases were reported. The present investigation was designed to overcome the omissions of previous studies, and examined the ability to read 46 single phonograms and 46 single ideograms aloud in four groups of sufficiently large numbers of patients; namely, seven pure alexics, 23 Broca aphasics, 13 Wernicke aphasics, and seven patients with alexia and agraphia. Ours are the first data to demonstrate unequivocally no consistent linkage between aphasia type and the patterns of impairment of phonogram and ideogram reading. The impairment patterns were not uniform across patients even in the same aphasia group. A majority of the cases in each group showed that phonograms and ideograms were unselectively impaired. However, ideogram reading was more difficult in three cases in the pure alexia and Broca aphasia groups, respectively, and in one case in the Wernicke aphasia group. Phonogram reading was more severely disturbed in four cases among the Broca aphasics and in one case among the patients with alexia with agraphia. An apparent variability of impairment patterns characterized the Broca aphasic group. These dyslexic patterns did not appear to correlate with the site and extent of lesions identified by computerized tomography scans. Past reports linking a particular impairment pattern of phonogram and ideogram reading and a specific lesion site were studies of single cases, and their conclusions seem oversimplified. While sensory and motor dysfunctions can usually be neuroanatomically localized in individuals, impairments of certain high cortical functions, such as the reading of phonograms and ideograms, may not be correlated with damage to definite neuroanatomical structures.


The influence of handedness on the distribution of muscular weakness of the arm in facioscapulohumeral muscular dystrophy.

The strength of 10 muscle groups in both arms was measured using hand-held myometry to determine the influence of handedness on left-right differences of muscle strength in facioscapulohumeral muscular dystrophy (FSHD). Two groups of subjects were studied: 24 healthy volunteers (19 right-handed), and 53 patients (42 right-handed) with autosomal dominant FSHD. An opposite left-right difference of strength of shoulder and arm muscles was found: right-handed volunteers were stronger on the right side, right-handed patients were stronger on the left side. This opposite left-right difference was statistically significant for the supraspinatus muscle, the wrist extensors and the shoulder internal rotators. The number of left-handed subjects was too small for statistical analysis. The relation between handedness and increased muscle weakness in right-handed FSHD patients suggests that mechanical factors may play a distinct role in the progression of muscle weakness in FSHD.


The evolution of neural circuits controlling feeding behavior in frogs.

Our approach to understanding motor systems is a phylogenetic, 'outside-in' approach, the goal of which is to identify behavioral transitions during phylogenesis and elucidate their neurological basis. In this paper, we review the results of recent behavioral, biomechanical and neurological studies on frog feeding behavior. These studies show that highly protrusible tongues have evolved numerous times independently among frogs, and that the biomechanics and neuromuscular control of feeding behavior have been transformed repeatedly during frog evolution. Many of the independent lineages possess unique biomechanical mechanisms for protracting their tongues and unique neural mechanisms for coordinating feeding behavior. In frogs, there has been considerable evolution at the interface between reticular central pattern generators (CPGs) associated with feeding and sensory feedback circuits that modulate feeding motor output. In particular, the roles of hypoglossal and glossopharyngeal sensory feedback appear to have been relatively plastic in their evolution. Prey-type dependence of hypoglossal sensory feedback in Rana suggests that the interaction between descending visual control and sensory feedback also may be evolutionarily plastic. Comparative studies have found that motor systems sometimes evolve conservatively across morphological and behavioral transitions (i.e., the shoulder in birds) or, alternatively, they may be subject to considerably more evolutionary change than is reflected in morphological characteristics (i.e., feeding in cichlids). We hypothesize that the CPG circuits for feeding behavior in the reticular formation may evolve conservatively because they are highly integrated, multifunctional networks which cannot be optimized for one function without compromising others. In contrast, the interfaces between the CPG, sensory feedback and descending control should be less constrained. When changes in motor patterns occur during evolution, it is likely that sensory feedback or descending control may be involved.


Chromatophore systems in teleosts and cephalopods: a levels oriented analysis of convergent systems.

The neural control of chromatophore display in cephalopod mollusks and teleost fishes is reviewed in the context of convergence of functional-anatomical pathways and mechanisms at several levels of organization. The effector elements or chromatophores are different in origin and design in the two groups of animals. Major functional differences appear to be in the speed of response (greatest in cephalopods) and the magnitude of non-neural control mechanisms (greatest in teleosts). Despite the differences, the elements demonstrate striking overall functional similarity. Elements of different types form highly organized array patterns of similar general complexity. Innervation patterns in cephalopods and teleosts seem comparable, with control being unidirectional (albeit in opposite directions); some elements demonstrate polyaxonal innervation. Motor units in both groups are generally composed of many chromatophores. Packard's concept of 'cronological units' of similar age-classes of chromatophores being innervated by similar age-classes of motor neurons greatly simplifies the understanding of relationships between the static arrays and the physiological units that utilize them to produce chromatic displays. The lower motor control areas for both groups have been grossly identified. Chromatomotor neurons in cephalopods are mostly located in the chromatophore lobes of the subesophageal brain while comparable systems in teleosts are situated in sympathetic chain ganglia (preganglionics) and the rostral spinal cord (postganglionics). Chromatic components are the simplest visually detectable units of color display, e.g. vertical bands and fin spots. They combine to form more complex chromatic patterns, which, in turn, are integrated with components of skin texture, posture and movement to produce display behaviors. Complexity of such systems seems to be of the same order of magnitude in both cephalopods and teleosts. Areas of the CNS related to each of the categorical levels have not been clearly defined. Crude patterning may take place in the basal and, perhaps, peduncle lobes in cephalopods and in the lower and intermediate medulla in teleosts. In both groups, higher level control relates to areas involved in sensorimotor integration and mediation of agonistic, sexual, and, perhaps, other types of behavior: the peduncle and optic lobes in cephalopods and the hypothalamus, tegmentum, otic tectum, torus semicircularis, thalamus and telencephalon in fishes. The systems appear to parallel each other in being organized hierarchically, with similar levels of complexity. Some of the regions may be especially important for regulating color patterns in response to visual input. Overall, chromatomotor control systems in cephalopods and teleosts demonstrate many apparent convergent features. Possible factors responsible for the similarities are discussed.(ABSTRACT TRUNCATED AT 400 WORDS)


Muscle architecture and control demands.

Muscles effect locomotion, and their gross architecture still poses analytical problems. These problems involve the arrangement of myofibers and motor units within muscles and that of muscles around joints. The arrangement of fibers may involve a range of considerations from the equivalence or nonequivalence of sarcomeres to placement, attachment, and angulation of fascicles and entire muscles; consequently, these levels and their development and coordination overlap. Many problems at the macroscopic level require clarification of how an animal uses a compartment of suite of muscles and whether morphological differences reflect functional ones. The understanding of intermediate architecture, including issues of compartmentation, pinnation, and concatenation, remains more elusive, as some morphologically distinct muscles may be functionally equivalent. As yet we have inadequate appreciation of the opportunities or limitations provided to the control system by a particular arrangement of fibers, or vice versa. Exploration of the rules that govern these conditions provides abundant opportunities for cooperation among neurobiologists, developmental biologists, physiologists and morphologists.


The spinal motor system in early vertebrates and some of its evolutionary changes.

Recent studies of the spinal motor systems of vertebrates allow us to begin to infer the organization of the motor apparatus of primitive vertebrates. This paper attempts to define some of the features of the motor system of early vertebrates based on studies of the motor systems in anamniotes and in Branchiostoma. It also deals with some changes in the primitive motor system during evolution. The primitive motor system consisted of myomeric axial muscles, with a functional subdivision of the musculature into non-spiking slow muscle fibers segregated in the myomeres from spiking fast ones. These fibers were innervated by two major classes of motoneurons in the cord-large motoneurons innervating faster fibers and small motoneurons innervating slow fibers. There was not a simple isomorphic mapping of the position of motoneurons in the motor column onto the location of the muscle fibers they innervated in the myomeres. Early vertebrates used these axial muscles to bend the body, and the different types of muscle fibers and motoneurons reflect the ability to produce slow swimming movements as well as very rapid bending associated with fast swimming or escapes. The premotor network producing bending was most likely a circuit composed of a class of descending interneurons (DIs) that provided excitation of ipsilateral motoneurons and other interneurons, and inhibitory commissural interneurons (CIs) that blocked contralateral activity and played an important role in generating the rhythmic alternating bending during swimming. This DI/CI network was retained in living anamniotes. At least two major descending systems linked the sensory systems in the head to these premotor networks in the spinal cord. The ability to turn on swimming by activation of DI/CI premotor networks in the cord resided at least in part in a midbrain locomotor region (MLR) that influenced spinal networks via projections to the reticular formation. Reticulospinal neurons were important not only for initiation of rhythmic swimming but also in the production of turning movements. The reticulospinal cells involved in turns produced their effects in part via monosynaptic connections with motor neurons and premotor interneurons, including some involved in rhythmic swimming. A prominent and powerful Mauthner cell was most likely present and important for rapid escape or startle movements. Some features of this primitive motor apparatus were conserved during the evolution of vertebrate motor systems, and others changed substantially. Many features of the early motor system were retained in living anamniotes; major changes occur among amniotes.(ABSTRACT TRUNCATED AT 400 WORDS)


Brainstem control of orienting movements: intrinsic coordinate systems and underlying circuitry.

A fundamental issue in the understanding of how the nervous system processes information is the way in which sensory information is used to initiate and guide movements. Recent progress has been made by taking an information processing approach in which information--for example, the spatial location of an object towards which an animal will orient--is tracked through the nervous system from sensory to motor levels. In this approach, neurally encoded information is characterized in terms of its representation within a neural or intrinsic coordinate system or set of neural coding parameters. For example, the retina codes spatial location in terms of the location of activity on the retinal surface, whereas motoneurons code spatial location in terms of the pulling directions of the muscles they activate. In between these two peripheral stages, the information passes through intermediate coordinate systems. These intermediate coordinate systems can be characterized by recording or altering the activity of small groups of neurons while an animal is performing a well-defined sensorimotor task. Spatial location information is used to guide orienting movements, those movements made by the eyes, ears, head, or body which function to center an object of interest in the animal's visual field. The optic tectum and forebrain, their connections to the medial mesencephalic and rhombencephalic brainstem tegmental cell groups, and subsequent connections to brainstem motor nuclei and spinal cord are employed to control fundamental aspects of this behavior. Studies reviewed herein indicate that following the retinotopic coding of spatial location in the retina and tectum, spatial location information appears to enter a different coordinate system at tegmental levels in which spatial aspects of orienting movement are coded in terms of their discrete horizontal and vertical components. This Cartesian coordinate system is an example of an abstract neural coordinate system, in that it is a simple, low-dimensional representation of spatial location which differs greatly from both sensory and motor representations. Also, this Cartesian representation may be common to many orienting movements, yet it appears to differ from the coordinate systems controlling other movement types such as stabilization or phasic movements. This suggests an hypothesis in which coordinate systems, especially at intermediate levels of processing, may be organized according to behavioral task as opposed to being determined by the particular sensory or motor system involved in the behavior. Understanding the evolutionary heritage and computational function of abstract neural coordinate systems, and the relation between different coordinate systems and behavioral tasks may be useful in understanding general aspects of sensory information processing and motor control.


p75 nerve growth factor receptor immunoreactivity in the human brainstem and spinal cord.

The distribution of nerve growth factor receptor (NGFR) immunoreactive profiles was investigated in the adult human brainstem and spinal cord using a monoclonal antibody directed against the primate low affinity (p75) NGFR. In the human brainstem, p75NGFR immunoreactive profiles were seen within the mesencephalic and descending nucleus of the trigeminal nerve, the nucleus and tractus solitarius, glossopharyngeal nerve, hypoglossal nucleus, nucleus subtrigeminalis, subnucleus ventralis of the central nucleus of the medulla, nucleus cuneatus and gracilis. At the level of the upper cervical spinal cord, p75NGFR immunoreactive profiles were also seen within the incoming dorsal roots, zone of Lissauer and substantia gelatanosa (lamina II). Virtually no immunoreactivity was associated with cervical spinal cord motor neurons. The demonstration of the p75NGFR in brainstem and spinal cord regions associated with the central transmission of peripheral sensory information suggests that these systems may be influenced by the trophic substance nerve growth factor.


VIP-like immunoreactivity in the suprachiasmatic nuclei of a mutant anophthalmic mouse.

The suprachiasmatic nuclei (SCN) of mutant anophthalmic (strain ZRDCT-An) and sighted control mice was examined using light microscopic methods for cytoarchitecture and immunocytochemistry for vasoactive intestinal polypeptide (VIP). The SCN of the anophthalmic mice were asymmetrical, and quite variable in cytoarchitecture. Immunocytochemistry for VIP revealed strong staining of cells and fibers. Within the SCN, the distribution of VIP-immunoreactive cells was more diffuse in anophthalmic mice than in the controls; immunoreactive neurons were frequently observed in ectopic locations in the mutant animals.


The specific N-methyl-D-aspartate (NMDA) receptor antagonist MK-801 blocks U-50,488, but not morphine antinociception.

The effect of the specific NMDA receptor antagonist MK-801 on antinociception produced by the kappa opiate receptor agonist U-50,488 and the mu receptor agonist morphine was assessed using the tail-flick test in rats. MK-801 (0.05 and 0.1 mg/kg) antagonized antinociception induced by all three doses of U-50,488 (7.5, 15 and 30 mg/kg), and potentiated antinociception induced by the lower (1 mg/kg) but not higher (5 mg/kg) dose of morphine. Naloxone at a dose of 1.0 but not 0.1 mg/kg blocked U-50,488 antinociception, indicating that MK-801 affects opiate antinociception. The present results are the first to suggest a critical role for the NMDA receptor in opiate antinociception involving the kappa receptor.


Electrophysiology of nitrous oxide on cerebellar granule cells: a single-cell study.

We recorded 18 single cells in the granule cell layer of the cat. Each single cell was screened and identified as a granule cell based on a set of criteria derived from known electrophysiological properties of granule cells. We then monitored the effects of nitrous oxide on the spontaneous activities and the auditory responses of these cells. Nitrous oxide consistently caused a severe inhibition of spontaneous activities as well as responses to sound in these cells. Furthermore, the amplitudes of their action potentials decreased during the inhibition. Three of the 18 cells were subsequently injected intracellularly with HRP. All three were verified to be granule cells. In those granule cells we recorded intracellularly, nitrous oxide did not change the resting membrane potentials. The gradual decrease in the amplitudes of action potentials suggested that some of the mechanisms leading to the genesis of action potentials were being altered by nitrous oxide. It is also possible that nitrous oxide may act on synaptic transmission at a site located postsynaptically on the granule cells.


Release of gamma-aminobutyric acid by visual stimulation in the kitten visual cortex.

Release of gamma-aminobutyric acid (GABA) was measured by brain microdialysis and high-performance liquid chromatography (HPLC) in the visual cortex of anesthetized kitten. The basal level of endogenous GABA release was 0.25 +/- 0.02 pmol/30 microliters dialysate (n = 8), which was near the lower limit of resolution of the present measuring system. When nipecotic acid, a GABA uptake inhibitor, was infused, release was increased 5-10 fold. The nipecotic acid-induced GABA output was not affected by the infusion of tetrodotoxin (TTX), a sodium channel blocker. Visual stimulation presented to one eye led to a marked increase in GABA output over the basal level. This effect was completely suppressed by TTX administration. These results suggest that the increase in GABA output in response to visual stimulation is due to an increase in GABAergic neuronal activity in the kitten visual cortex.


Peripheral origin of olfactory nerve fibers by-passing the olfactory bulb in Xenopus laevis.

After DiI injections into the diencephalon of Xenopus, two types of retrogradely labelled cells were found in the nasal area: (i) receptor cells in the olfactory epithelium and (ii) a small cell group located between the main olfactory epithelium and the vomeronasal system. These results reveal an extensive extrabulbar olfactory projection of olfactory receptor cells. Fibers of these cells do not terminate in the olfactory bulb but innervate targets in the diencephalon directly. The other type of retrogradely labelled cells, apparently, are not part of any epithelium. They resemble similar cell groups which have previously been regarded as part of the nervus terminalis system in other vertebrates.


Hippocampal mossy fiber distribution does not correlate with two-way active avoidance performance in backcross lines derived from inbred mouse strains DBA/2 and C3H.

To elucidate the relation of genetically induced variations in two-way active avoidance performance and the extent of the hippocampal infra/intrapyramidal mossy fiber system (IIP-Mf) in inbred mouse strains DBA/2 (high performance, small IIP-Mf) and C3H (low performance, large IIP-Mf) we investigated a backcross line containing the DBA/2-derived genetic determinants for high avoidance performance in a C3H background. In the seventh backcross generation variations in performance analogous to the original strains were observed whereas the extent of mossy fibers was nearly homogeneous. This result suggests that both variables are determined by separate genetic factors at least in these strains and that differences in mossy fiber distribution are not an essential prerequisite for different active avoidance performance.


Olfactory ensheathing cells myelinate dorsal root ganglion neurites.

The objective of this study was to determine whether olfactory ensheathing cells (from E18 rat embryos) could myelinate dorsal root ganglion neurites in vitro. By four weeks many of the S100-positive ensheathing cells were Gal-C+ and MBP+ and had begun to myelinate the larger axons, as visualized with the electron microscope. We conclude that olfactory ensheathing cells can form a myelin sheath if given the opportunity to ensheath neurites of a proper size.


Parvalbumin and calbindin D-28K in the human entorhinal cortex. An immunohistochemical study.

Research is here reported on the distribution of immunoreactivities of the calcium-binding proteins parvalbumin and calbindin D-28K in the entorhinal cortex of normal human brains. Topographically, parvalbumin immunoreactive neurons were only seen in the lateral portion of the rostral entorhinal cortex, in continuity with the adjacent perirhinal cortex. The intermediate and caudal portions gave positive results along the mediolateral extension of the entorhinal cortex. The laminar distribution of parvalbumin immunoreactive neurons was similar throughout the entorhinal cortex. Heavy immunostaining, largely coincident with cell islands, was observed in cells and fibers in layer II, being densest in the deep half of layer III and more sparsely distributed in layers V and VI. Calbindin D-28K immunoreactivity was found throughout the entorhinal cortex. In contrast to parvalbumin immunoreactivity, calbindin D-28K was present from layer I up to upper layer III, the neurons being most numerous in the cell islands of layer II. These results show that rostromedial portions of the human entorhinal cortex contain calbindin immunoreactivity, but not parvalbumin, while the lateral, intermediate and caudal portions of the entorhinal cortex contain both calcium-binding proteins. As it is known that these two proteins belong to a subset of GABAergic neurons, we suggest that a topographical diversity in some of the cells may be responsible for inhibitory effects in the human entorhinal cortex. This proposed diversity might be relevant to the processing of information that the entorhinal cortex conveys to the dentate gyrus and receives from various components of the hippocampus, the subicular complex and other cortical and subcortical sources.


The basis and functional role of the late EMG activity in human forearm muscles following wrist displacement.

The present paper examines the hypothesis that the long latency EMG activity produced by muscle stretch is the result of long loop reflex pathways involved in the control of limb stiffness. We recorded the cerebral responses and late EMG activity in agonist and antagonist muscles following sudden stretch of the wrist extensor muscles under 4 experimental conditions in 11 subjects. In each experiment subjects held their right wrist extended isometrically against a constant force of 2.3 N and a trial was begun with a step increase in the force from 2.3 N to 18.4 N, to stretch the extensor muscle. In the first and second experiments the force change occurred unpredictably and subjects had to either oppose the perturbation (Unpredictable Oppose) or relax the forearm muscles once the increase in force was detected (Unpredictable Let-Go). In the third and fourth experiments the force change occurred predictably when subjects pressed a thumb switch with the left hand to cause it. As before, subjects were instructed to either oppose the perturbation (Predictable Oppose) or relax the forearm muscles (Predictable Let-Go). Responses were recorded from the flexor and extensor carpi radialis muscles and from the scalp. When the perturbing force occurred unpredictably, early latency EMG activity (the MI response) was seen in the stretched extensor muscle, and longer latency EMG activity was seen simultaneously in both extensor and flexor muscles. When the force change occurred predictably the late EMG activity was considerably attenuated, especially in the Predictable Let-Go condition. Cerebral responses similarly depended upon the predictability of the perturbation.(ABSTRACT TRUNCATED AT 250 WORDS)


Subcellular distribution of serotonin in the lamprey spinal cord.

The subcellular distribution of serotonin (5-hydroxytryptamine; 5-HT) in the lamprey (Ichtyomyzon unicuspis, Lampetra fluviatilis) spinal cord was investigated by using ultracentrifugation on continuous density gradients combined with an electron microscopic analysis of the gradients and of immunostained tissue. Endogenous 5-HT was analyzed by high-performance liquid chromatography with electrochemical detection. After differential centrifugation, the highest levels of 5-HT were found in the particulate fractions. After ultracentrifugation of lysed synaptosomal fractions on continuous sucrose gradients and the subsequent sedimentation of the individual fractions, 5-HT showed a biphasic distribution in the gradient. The two peaks corresponded to 0.30-0.40 M and 0.85-1.05 M sucrose. Electron microscopy of intact tissue showed that some of the boutons were strongly immunoreactive to 5-HT with dense precipitates over large granular vesicles. The area around these large vesicles, however, also showed reaction product. Large granular vesicles could be clearly distinguished in the immunostained axonal varicosities. In tissue not processed for 5-HT immunoreactivity it was seen that the varicosities contained not only large dense-cored vesicles, but also small agranular vesicles. An electron microscopical analysis of the subcellular fractions revealed that the fraction corresponding to the "light" 5-HT peak contained numerous vesicular structures, which in most cases were electron lucent. In the "heavy" fractions, nerve ending particles containing vesicles of various sizes were observed. The results suggest that 5-HT in the lamprey spinal cord may be distributed in more than one subcellular compartment which, apart from the cytosol, possibly corresponds to small and large synaptic vesicles.


N-methyl-D-aspartate unilaterally injected into the dorsal striatum of rats produces contralateral circling: antagonism by 2-amino-7-phosphonoheptanoic acid and cis-flupenthixol.

To evaluate the possible contribution of dorsal striatal glutamate receptors to motor behavior, circling responses were observed in rats following unilateral intrastriatal microinjections of the agonist, N-methyl-D-aspartate (NMDA) or the antagonist, 2-amino-7-phosphonoheptanoic acid (APH). The role of dopamine (DA) in NMDA-produced circling also was evaluated. In experiment 1, an NMDA dose of 5.0 micrograms (in 0.5 microliter), but not 0.5 or 0.05 microgram produced significant contraversive circling. In experiment 2, an APH dose of 10.0 micrograms but not 1.0 or 0.1 microgram produced significant ipsiversive circling. In experiment 3, microinjection of the ineffective 0.1 microgram dose of APH or a dose (20 micrograms) of the DA antagonist, cis-flupenthixol, that did not produce circling when administered alone, significantly reduced the circling response produced by the 5.0 micrograms dose of NMDA. As NMDA produced circling in the same direction as that seen following similar unilateral injections of locomotion-stimulating DA agonists, the present results suggest that glutamate, acting via NMDA receptors in the dorsal striatum, may exert an excitatory influence on motor systems. The observation that a DA receptor blocker antagonized the NMDA response further suggests that the observed motor excitatory effect of glutamate at NMDA receptors requires concurrent stimulation of DA receptors in the same region of the striatum.


5-Hydroxytryptamine-3 receptors modulate synaptic activity in the rat nucleus tractus solitarius in vitro.

Whole-cell patch clamp recordings were made from neurons in the rat nucleus tractus solitarius (NTS) in transverse brainstem slices. 5-Hydroxytryptamine (5-HT, 100 microM) and the selective 5-HT3 receptor agonist 2-methyl-5-HT (2-CH3-5-HT, 100 microM) depolarized 86% of NTS neurons at resting membrane potential (Vm). This response was resistant to tetrodotoxin (TTX) and Co2+ application. In addition, 2-CH3-5-HT (500 nM-100 microM) increased the amplitude and frequency of both excitatory and inhibitory spontaneous synaptic potentials. This effect was also TTX-resistant, but was abolished by Co2+. The effects of 2-CH3-5-HT on EPSPs and IPSPs evoked by electrical stimulation of the tractus solitarius (TS) were analyzed separately in the presence of bicuculline or 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), respectively. Concentrations of 2-CH3-5-HT between 500 nM and 1 microM decreased the amplitude of evoked EPSPs and IPSPs with similar potency. The selective 5-HT3 receptor antagonists ICS 205-930 (10 nM) and MDL 72222 (10 microM) reversibly blocked the effects of 2-CH3-5-HT at all doses examined. It is concluded that 5-HT3 receptors can mediate both pre- and postsynaptic responses in the NTS.


Responses of globus pallidus neurons to cortical stimulation: intracellular study in the rat.

The responses of globus pallidus (GP) neurons to stimulation of the sensorimotor cortex, the neostriatum, and the subthalamic nucleus were intracellularly recorded in anesthetized rats. Stimulation of the cortex evoked a sequence of postsynaptic responses including an initial short EPSP, a short IPSP, and a late EPSP with multiple spikes in most of the repetitively firing GP neurons. The response pattern was very similar to those evoked by striatal stimulation, except that the latencies were longer. An acute knife cut placed immediately caudal to the substantia nigra caused no significant change in the responses to cortical and striatal stimulation. Stimulation of the subthalamic nucleus evoked a short latency EPSP overlapped with an IPSP. The polarity of all the IPSPs was reversed by a Cl- injection. A systemic injection of picrotoxin abolished all the IPSPs and unmasked large depolarizations with multiple spikes. An ibotenic acid lesion of the subthalamic nucleus eliminated both the initial short latency and late EPSPs to cortical and striatal stimulation and disclosed a prominent IPSP. Stimulation of the lesioned subthalamic nucleus also evoked large, short latency IPSPs without noticeable EPSPs. These results indicate that (i) the IPSPs evoked by cortical, striatal, and subthalamic stimulation were mediated by a GABAA receptor, (ii) both the initial and late EPSPs to cortical and striatal stimulation involved activation of the subthalamic nucleus but not brainstem nuclei, and (iii) cortically derived signals mediated through the neostriatum (i.e. long latency IPSPs) and the subthalamic nucleus (i.e. short latency EPSPs) converged on most GP neurons.


Noradrenergic activation in the paraventricular nucleus during acute and chronic immobilization stress in rats: an in vivo microdialysis study.

In vivo microdialysis was used to study the effects of single (2 h) or repeated (2 h for 7 consecutive days) immobilization (IMMO) stress on extracellular fluid concentrations of norepinephrine (NE) and the deaminated metabolites of NE and dopamine, dihydroxyphenylglycol (DHPG) and dihydroxyphenylacetic acid (DOPAC) in the paraventricular nucleus of conscious rats. During IMMO, NE, DHPG, and DOPAC levels increased markedly, with similar peak values and time courses in the repeatedly stressed and previously unstressed groups. NE levels during a 2-h baseline period were lower in the repeatedly stressed group than in the unstressed group (99 +/- 9 pg/ml vs. 167 +/- 13 pg/ml, P less than 0.05), whereas DHPG (1,697 +/- 263 pg/ml vs. 1,424 +/- 194 pg/ml) and DOPAC (5,989 +/- 863 pg/ml vs. 4,428 +/- 1150 pg/ml) levels tended to be higher, so that the NE/DHPG ratio at baseline was significantly lower in the repeatedly stressed group (P less than 0.05). The results indicate that IMMO stress enhances NE release, reuptake, metabolism, and synthesis in the PVN. Repeated exposure to IMMO may decrease the microdialysate NE/DHPG ratio by inhibiting exocytotic release or enhancing neuronal reuptake of NE. In either case, the results suggest that repeated exposure to stress alters the release and disposition of NE in the PVN of conscious animals.


Excitatory effects of thyrotropin-releasing hormone on neurons within the nucleus ambiguus of adult guinea pigs.

The electrophysiological effects of thyrotropin-releasing hormone (TRH) on neurons within the nucleus ambiguus (NA) of adult guinea pigs were studied using an in vitro brain stem slice preparation. In 0.01-1.0 micron TRH, NA neurons depolarized (25/39), expressed enhanced postinhibitory rebound (8/8 tested), or exhibited oscillations of the membrane potential (17/39). Because the amplitude of postinhibitory rebound in tetrodotoxin (TTX) at various membrane potentials was not altered by TRH, it suggests that TRH enhanced postinhibitory rebound indirectly by depolarizing the cell membrane. The membrane potential oscillations in NA neurons were persistent in TTX and their frequency was dependent on the membrane potential, suggesting that these oscillations were due to intrinsic membrane properties and not to synaptic inputs. The excitation of NA neurons in vitro by TRH suggests that endogenous TRH may modulate the activity of neurons involved in the regulation of respiratory and autonomic function.


Suppression of interictal bursting in hippocampus unleashes seizures in entorhinal cortex: a proepileptic effect of lowering [K+]o and raising [Ca2+]o.

The relation between interictal bursts (IIBs) and seizures in epilepsy is obscure. Results from some human and animal studies suggest that IIBs may actually suppress seizure activity. This appears particularly true in the zero magnesium in vitro seizure model. Here we provide new evidence in support of this and new insight into the mechanisms of seizure suppression in this model. Brain slices containing hippocampus and entorhinal cortex were bathed in zero magnesium medium. Electrographic seizures appeared, then were replaced by IIBs. Upon lowering [K+]o and raising [Ca2+]o the IIBs disappeared and the seizures reappeared. Repeated stimuli mimicking IIBs then suppressed seizures again. Selective knife cuts revealed that the IIBs originated in the hippocampus (area CA3) whereas the seizures originated in entorhinal cortex. These results confirm that IIBs suppress seizures in the zero magnesium model. They also show that an important aspect of the interaction between IIBs and seizures in this model is the anatomical segregation of their respective sites of origin. This may apply in other models and in human epilepsy as well. Finally, these results illustrate that one consequence of the anatomical segregation and mutual interaction of IIBs and seizures is that influences which are locally pro- or antiepileptic can have opposite effects in a broader region.


Gamma-vinyl GABA prevents hippocampal and substantia nigra reticulata damage in repetitive transient forebrain ischemia.

GABAergic inhibitory mechanisms may offer protection to neurons after global ischemia. We tested the effects of gamma-vinyl GABA, a GABA-transaminase inhibitor, via continuous infusion in the third ventricle (Alza pumps) in a gerbil model of repetitive forebrain ischemia. We used two episodes of 3 min duration with a 'reperfusion' interval of 1 h between the insults. Histological analysis was done with silver staining 5 days after the insult. Our results show that there is significant protection of the hippocampus CA1 region and substantia nigra reticulata in treated animals compared to controls. An increase in GABA levels, decrease in glutamate, or mild hypothermia, may be potential mechanisms for this protection. GABAergic agents may prove useful agents in repetitive ischemia.


Studies related to the use of colchicine as a neurotoxin in the septohippocampal cholinergic system.

Colchicine has been shown to be neurotoxic to cholinergic neurons in the medial septum 1 week following intracerebroventricular injections. The experiments described here were designed to examine the selectivity of this effect over a longer time course, and to examine the role of axoplasmic transport in the neurotoxic effect. As previously reported, 1 week after intracerebroventricular injections of colchicine, the numbers of choline acetyltransferase (ChAT)-immunoreactive neurons in the medial septum-diagonal band complex (MSDB) were reduced to 38% of control; this reduction was stable 2 and 3 weeks post injection. Injections of colchicine placed into the body of the fornix produced similar results. GAD-immunoreactive somata, the other major population of neurons in the MSDB, were unaffected 3 weeks following colchicine, as previously reported 1 week following similar injections. The normal AChE staining pattern in the hippocampus, particularly the dentate gyrus, was depleted following either ICV or intrafornical injections of colchicine. This depletion was more severe with longer survival times. Injections of lumicolchicine, an isomer of colchicine which does not bind tubulin, had no effect on ChAT-immunoreactive neurons in the MSDB or on AChE staining in the hippocampus. Injections of colchicine, but not of lumicolchicine, partially blocked the retrograde transport of the fluorescent dye Fluoro-Gold from the hippocampus to the MSDB. In addition, the content of NGF in the hippocampus rose 84% above control values 2 weeks following colchicine and remained elevated at three weeks. Together these results indicate that colchicine is selectively toxic for cholinergic neurons in the septohippocampal system, and suggest that the alkaloid's neurotoxic effects work via the blockade of axoplasmic transport.


Electrophysiological effects of MK-801 on rat nigrostriatal and mesoaccumbal dopaminergic neurons.

The electrophysiological effects of the non-competitive N-methyl-D-aspartate (NMDA) antagonist (+)-MK-801 (MK-801) on nigrostriatal and mesoaccumbal dopaminergic (DA) neurons were evaluated in chloral hydrate-anesthetized rats. MK-801 (0.05-3.2 mg/kg, i.v.) stimulated the firing rates of 14 (74%) of 19 nigrostriatal DA (NSDA) neurons and all 16 mesoaccumbal DA (MADA) neurons tested. Stimulatory effects of the drug were more prominent on MADA neurons. Interspike interval analysis revealed that MK-801 also regularized DA neuronal firing pattern. Acute brain hemitransection between the midbrain and forebrain attenuated the stimulatory effects of MK-801 on firing rate and blocked the effects on firing pattern. Similar to MK-801, hemitransection itself increased NSDA and MADA cell firing rates and regularized firing pattern. Both i.v. and iontophoretic MK-801 blocked the excitatory effects of iontophoretic NMDA but did not affect excitations caused by the non-NMDA glutamatergic receptor agonists quisqualate and kainate. Iontophoretic MK-801 had no effect alone. These results suggest that the excitatory effects of i.v. MK-801 on DA neuronal activity are not due to direct actions on DA neurons. Glutamatergic projections originating anterior to the hemitransection appear to play a role in the effects of MK-801 on DA neuronal activity.


Neurons of the chick brain and retina expressing both alpha-bungarotoxin-sensitive and alpha-bungarotoxin-insensitive nicotinic acetylcholine receptors: an immunohistochemical analysis.

Immunohistochemical methods were used to study the possible co-localization of two alpha-bungarotoxin-sensitive (alpha 7 and alpha 8) and two alpha-bungarotoxin-insensitive (beta 2 and alpha 3) subunits of the nicotinic acetylcholine receptors in neurons of the chick brain and retina. Several structures contained neurons that were doubly-labeled with antibodies against the alpha 7 subunit and the beta 2 subunit. These structures included, for example, the interpeduncular nucleus, nucleus spiriformis lateralis, optic tectum, pretectal visual nuclei, and the lateral hypothalamus. Double-labeling with antibodies against the alpha 7 and alpha 8 subunits was also seen in several regions, which included the interpeduncular nucleus, visual pretectum, lateral hypothalamus, dorsal thalamus, and the habenular complex. In the retina, many cells in the inner nuclear layer were observed to contain alpha 8 and alpha 3 subunits, whereas neurons in the ganglion cell layer were seen to contain alpha 7 and alpha 8 or, less frequently, alpha 7 and alpha 3 subunits. These results indicate that alpha-bungarotoxin-sensitive and alpha-bungarotoxin-insensitive subunits of the nicotinic receptors are co-expressed by neurons of the chick brain and retina.


Transport, uptake, and metabolism of blood-borne vasopressin by the blood-brain barrier.

Transport, binding, and metabolism of [phenylalanyl-3,4,5,-3H(N)]arginine vasopressin (AVP) by the blood-brain barrier (BBB) was studied in adult guinea-pigs by means of a novel vascular brain perfusion (VBP)/capillary depletion technique and HPLC. A time-dependent, progressive brain uptake of 3H-radioactivity was measured over the 10 min period of VBP both in brain homogenates and in brain tissue depleted of cerebral microvessels. The unidirectional blood-to-brain transport constant, K(IN), estimated by multiple-time tissue uptake analysis of the homogenate and postcapillary supernatant, indicated that the BBB transfer rate for [3H]AVP (K(IN) = 2.37 +/- 0.25 microliters min-1 per gram brain homogenate) was almost 10 times higher than for simultaneously perfused [14C]sucrose, a cerebrovascular space marker. In contrast to homogenate and postcapillary supernatant, the [3H]radioactivity determined in the vascular pellet after dextran density centrifugation of the brain homogenate was very low and only somewhat higher than for [14C]sucrose. HPLC analysis of the perfused brain tissue revealed time-dependent degradation of the blood-borne neuropeptide. The percentage of intact [3H]AVP as determined in the postcapillary supernatant progressively declined during brain perfusion, from 49% at 1 min to 11.9% at 10 min. The major detectable labeled metabolite was [3H]phenylalanine, the labeled amino acid residue of [3H]AVP. The aminopeptidase inhibitor bestatin (0.5 mM), perfused simultaneously with [3H]AVP by the VBP technique, did not alter tissue uptake of [3H]AVP, indicating that there was no significant hydrolysis of peptide by the luminal BBB surface.(ABSTRACT TRUNCATED AT 250 WORDS)


Effect of subcutaneous administration of the chemical algogen formalin, on 5-HT metabolism in the nucleus raphe magnus and the medullary dorsal horn: a voltammetric study in freely moving rats.

The effect of subcutaneous administration of the chemical algogen formalin, on serotonin (5-HT) metabolism in the nucleus raphe magnus (NRM) and the medullary dorsal horn (MDH) has been investigated using in vivo 5-hydroxyindole electrochemical (peak '3') detection with treated, multi-carbon fiber electrodes and differential pulse, or normal pulse, voltammetry in freely moving rats. The subcutaneous (s.c.) injection of 50 microliters of 10% formalin in the left forepaw was followed, at the NRM level, by a significant increase in the voltammograms as compared to controls (50 microliters of saline 0.9% s.c. in left forepaw) for about 70 min after the injection, before a return to control values. At the MDH level, the formalin injection induced no significant effect on peak 3, as compared to controls, during the first 70 min. After that, the voltammograms significantly increased and remained above controls for up to 180 min. Thus, the time-courses of NRM and MDH effects appear markedly different. These findings suggest that, depending on the anatomical level (NRM or MDH) and/or the period of observation, one can measure differences in the time-course of the increase in 5-HT metabolism in the NRM-dorsal horn serotonergic system by tonic noxious stimuli, such as the formalin test.


Blood-brain barrier integrity and brain water and electrolytes during hypoxia/hypercapnia and hypotension in newborn piglets.

This study examines the effects of hypoxia/hypercapnia and hypoxia/hypercapnia with hypotension (hypotensive-hypoxia/hypercapnia) on blood-to-brain transfer constants (K1) for sodium and mannitol and brain water and electrolyte contents in newborn piglets. Hypoxia/hypercapnia was induced for 60 min with the piglets breathing a gas mixture of 15% carbon dioxide, 10-12% oxygen, and 73-75% nitrogen adjusted to achieve an arterial pH less than 7.15, pO2 less than 40, and pCo2 greater than 60 mmHg and hypotension for 20 min by rapid phlebotomy to achieve a mean arterial blood pressure less than 40 mmHg. Piglets were studied during 1 h of, and 24 h after resuscitation from hypoxia/hypercapnia (arterial pH 6.9 +/- 0.18, pO2 36 +/- 6 mmHg, pCO2 68 +/- 8 mmHg, mean +/- S.D.) and 10 min, and 24 h after resuscitation from hypotensive-hypoxia/hypercapnia (mean arterial blood pressure 28 +/- 10 mmHg, mean +/- S.D.). Values for K1 for sodium and mannitol, measured using the integral technique were 15.9 and 5.2 ml.g-1.min-1 x 10(4) respectively, in 2-4-day-old controls, suggesting that the barrier is fully developed in newborn piglets. Values were not different during or after hypoxia/hypercapnia or 24 h after hypotensive-hypoxia/hypercapnia. Ten to forty min after hypotensive-hypoxia/hypercapnia, there was a proportional decrease in the K1 for sodium and mannitol of about 40%. These results suggest that the newborn piglet is similar to the adult with respect to impermeability of the blood-brain barrier to ions and small molecules and resistance of this barrier to systemic hypoxia/hypercapnia and hypotension.(ABSTRACT TRUNCATED AT 250 WORDS)


Amastatin potentiates drinking elicited by osmotic stimuli: evidence for peptidergic mediation of intracellular dehydration-induced thirst.

Aminopeptidase inhibitors have been demonstrated to enhance the behavioral effects of both exogenously applied and endogenously released neuropeptides. In this study peptidase inhibitors were used as probes for involvement of central neuropeptides in osmotically-induced drinking behavior. Intracerebroventricular (i.c.v.) injections of amastatin, an aminopeptidase A inhibitor, potentiated water intake induced by subcutaneous injections of hypertonic saline. Drinking responses to i.c.v. infusions of hypertonic saline were also enhanced when amastatin was added to the infusions. The effect was not attenuated by the angiotensin receptor antagonist, [Sar1, Thr8]angiotensin II, which suggests that angiotensins do not play a role in the over-drinking. Drinking responses to centrally infused hypertonic saline were not enhanced by i.c.v. thiorphan, an endopeptidase inhibitor; this provides evidence that the effects of amastatin are specific for a particular class of peptidases. These results suggest that there is a role for an endogenous, non-angiotensinergic brain peptide in the mediation of osmotic thirst.


Behavioral effects of mouse interferons-alpha and -gamma and human interferon-alpha in mice.

Behavioral effects of murine interferon-alpha and -gamma were explored using a well-characterized system for detecting interferon effects. In addition, the effectiveness of human interferon-alpha was determined. Mouse gamma-interferon decreased activity and food-related behavior, effects that were similar to the effects of mouse alpha-interferon. Equivalent doses of the human preparation had little effect upon these measures in mice. Some common action of alpha- and gamma-interferon is likely responsible for the similar effects seen for the two molecules. Low effectiveness of the human preparation in mice was predicted due to the differences in receptors between species.


Pulse jitter retards kindling without affecting brain-stimulation reward.

Convulsions in rats bar-pressing for electrical stimulation of the brain can interfere with the behavioural evaluation of the properties of the reward substrate. If seizure activity involves the synchronous activation of populations of neurons, then the regular timing of pulses may be important in epileptogenesis produced by electrical stimulation. In the first experiment we found that behaviourally derived, charge-duration functions collected from three self-stimulating rats with lateral hypothalamic electrodes were not affected by introducing a controlled, 50% jitter to the interpulse interval of self-stimulation trains. In the second experiment, we kindled two groups of 6 rats with once-daily, 1.0 s bursts of either regularly timed or jittered pulses to the amygdala until full motor seizures developed. Although both groups eventually attained Class 5 convulsions, the time to first seizure was about two days longer with jittered pulses. Thus, while the circuitry that accumulates input from self-stimulation is not affected by the irregularity of asynchronous pulses, the jitter does seem to retard the development of seizures.


Subthalamic nucleus cell firing in the 6-OHDA-treated rat: basal activity and response to haloperidol.

Single unit recordings from neurons of the subthalamic nucleus were made in control and 6-hydroxydopamine (6-OHDA)-treated rats anesthetized with chloral hydrate. Subthalamic nucleus cells in this preparation exhibited a wide range of firing rates and three different firing patterns. These patterns were defined as 'burst', 'normal', and 'mixed' based on comparisons of their interspike interval histograms. Four to 6 weeks after 6-OHDA treatment there was no change in the basal firing rates of subthalamic nucleus cells, but there was a significant shift in firing pattern, with a smaller proportion of cells exhibiting the 'normal' firing pattern. The response of subthalamic nucleus neurons to acute administration of haloperidol was also altered in 6-OHDA-treated rats tested 4-6 weeks post-lesion, with a significantly greater proportion of cells responding to doses of haloperidol as low as 0.2 mg/kg (i.v.) with increases in firing rate of 20% or more. These results suggest that the subthalamic nucleus is probably not involved in the increases in basal levels of dopamine cell activity observed previously in the 6-OHDA-treated rat, but may play a role in the acute induction of depolarization block of dopamine cell firing in response to haloperidol administration in this model.


Changes in D2 but not D1 receptor binding in the striatum following a selective lesion of striatopallidal neurons.

We have used the immunotoxin OX7/saporin, a suicide transport agent, to selectively lesion striatopallidal neurons. Following injection of OX7/saporin into the globus pallidus, in situ hybridization for preproenkephalin mRNA was examined in the striatum to confirm successful retrograde neurotoxicity. Comparison of D1 and D2 receptor binding in the striatum demonstrated that D2 but not D1 receptor binding sites are localized to striatopallidal neurons.


Amelioration of ischemic spinal cord damage by postischemic treatment with propentofylline (HWA 285).

The effect of the xanthine derivative propentofylline (HWA 285) on metabolic and functional recovery in rabbit spinal cord after 20 and 30 min ischemia and 4 days of reperfusion was investigated. Pre-treatment with 20 mg/kg significantly improved recovery of the energy state in the spinal cord, however, without significant functional recovery of hindlimbs. In contrary, post-treatment with HWA 285 recovered the energy state to pre-ischemic value and also significantly improved functional recovery. These findings suggest that the neuroprotective mechanism of HWA 285 in the spinal cord is not associated with inhibition of glutamate release as supposed to operate in the gerbil brain.


Optic nerve transection decreases 2-[125I]iodomelatonin binding in the chick optic tectum.

The distribution of specific 2-[125I]iodomelatonin binding sites in the various layers of the chick optic tectum was analyzed using quantitative receptor autoradiography. Following unilateral optic nerve transection, binding in the optic fiber layer and superficial retinorecipient layers of the contralateral tectum was significantly decreased at 7 and 14 days, but not at 1 day, following transection. The results are consistent with the presence of presynaptic melatonin receptors on axon terminals of retinotectal fibers.


Preservation of energy metabolites by carnitine in the mouse brain under ischemia.

Ischemia (decapitation) caused marked changes of energy metabolites in the mouse brain. However, the changes were clearly less severe, when the mouse was pre-treated with L-carnitine. This suggests that L-carnitine may protect the brain from the ischemic attack. Since D-carnitine showed a similar effect, the effects may be due to mechanism(s) other than the 'classical' function of carnitine.


Pre-Bötzinger complex in cats: respiratory neuronal discharge patterns.

Patterns of respiratory neuronal discharge in the pre-Bötzinger complex, hypothesized as a brainstem site generating respiratory rhythm, are described in adult cats. Signals were recorded from neurons in the Bötzinger complex, pre-Bötzinger complex and rostral ventral respiratory group (rVRG) of anesthetized adult cats. The pre-Bötzinger complex, located caudal to expiratory-modulated Bötzinger neurons, contained a mix of neurons with inspiratory-modulated, expiratory-modulated, or phase-spanning patterns of impulse activity, in contrast to the more homogenous neuronal distributions characteristic of adjacent Bötzinger and rVRG regions.


Comparison of the effects of NMDA antagonists on medial vestibular nucleus neurons in brainstem slices from labyrinthine-intact and chronically labyrinthectomized guinea pigs.

The responses of ipsilateral medial vestibular nucleus (MVN) neurons in brainstem slices from guinea pigs compensated for a unilateral labyrinthectomy (UL), to the N-methyl-D-asparate (NMDA) receptor/channel antagonists CPP and MK801, were compared with those of MVN neurons in brainstem slices from labyrinthine-intact guinea pigs observed in a previous study. The average resting activity of ipsilateral MVN neurons from compensated animals was significantly higher than that for MVN neurons from labyrinthine-intact animals; however, there were no significant differences in the average magnitude of the decrease in firing rate from baseline in response to CPP or MK801 and the only significant difference in the number of responses was to MK801, where fewer ipsilateral MVN neurons from compensated animals responded with a decrease in firing rate. These results suggest that vestibular compensation is not associated with an up-regulation or increased affinity of NMDA receptors in the MVN ipsilateral to the UL.


Selective activation of mesoamygdaloid dopamine neurons by conditioned stress: attenuation by diazepam.

Populations of dopamine (DA) neurons in the rat brain are selectively activated by stress, and the response is attenuated by the administration of anxiolytics. Given the role of the component nuclei of the amygdaloid complex in conditioned associations, stress responses and the anxiolytic effects of benzodiazepines, we hypothesized that particular mesoamygdaloid DA projections might be especially sensitive to the effects of conditioned stress and to diazepam (DZ). We mapped the effect of a conditioned stressor on the concentration of the DA metabolite homovanillic acid (HVA) in distinct amygdaloid nuclei and other brain nuclei and areas and the effect of DZ (1 or 3 mg/kg) on the conditioned response in drug-experienced subjects. The conditioned stress paradigm resulted in significant elevations in classical indices of stress, including serum corticosterone and plasma epinephrine. Conditioned stress-induced increases in the estimated activity of DA neurons were specific for DA neurons projecting to the central, basolateral and lateral amygdaloid nuclei, and for DA projections to the dorsal septal nucleus. Conditioned stress-induced increases in the HVA concentration of responsive amygdaloid nuclei were antagonized by low, anxiolytic doses of DZ. These results indicate a role for a subset of mesoamygdaloid DA projections in transducing the impact of perceived stressors on the output of the amygdaloid complex. A role for particular amygdaloid DA projections in the formation of conditioned fear or anticipatory anxiety and its modulation by anxiolytics is also suggested.


Protective effects of serotonin reuptake inhibitors, citalopram and clomipramine, against hippocampal CA1 neuronal damage following transient ischemia in the gerbil.

To clarify the role of serotonin in cerebral ischemia, we examined the effects of selective serotonin reuptake inhibitors, citalopram and clomipramine, on ischemic neuronal damage in the gerbil. Pretreatment with citalopram (40 mg/kg i.p.) and clomipramine (20 mg/kg i.p.) protected against neuronal destruction of hippocampal CA1 pyramidal cells following 5 min of forebrain ischemia. Furthermore, microdialysis assays showed that a striking increase in extracellular excitatory amino acid levels during ischemia was significantly inhibited by pretreatment with citalopram and clomipramine. However, citalopram (40 mg/kg i.p.) did not alter the extracellular amino acid concentrations in normal gerbils. Thus, serotonin reuptake inhibitors have a protective effect against ischemic neuronal damage. Furthermore, the present result suggests that the protective effect is mediated through prevention of the accumulation of extracellular excitatory amino acids during and after ischemia.


Immunocytochemical localization of 205 kDa microtubule-associated protein (205 kDa MAP) in the guinea pig organ of Corti.

We have studied the immunocytochemical localization of microtubule-associated proteins (MAPs) in the guinea pig organ of Corti. Using immunological methods with antibodies against MAP1A, MAP1B, MAP2, tau and 205 kDa MAP, we have identified 205 kDa MAP as a major MAP of the sensory epithelium in the organ of Corti. Immunoperoxidase microscopic study has shown that both cochlear hair cells and supporting cells reacted with anti-205 kDa MAP antibody. Immunoelectron microscopy revealed that 205 kDa MAP was associated with most microtubules in the sensory epithelial cells. It was also associated with the microtubules of bundle structures within supporting cells, suggesting that this MAP might form a part of cross-bridges between microtubules and between microtubules and actin filaments in the bundle structure. In contrast, MAP1A, MAP1B and tau, which are known to be expressed in neuronal tissue, were localized only in nerve fibers in the organ of Corti, not in the sensory epithelium. MAP2, which is known to be localized in dendrites and soma of nerve cells, was not distributed in nerve fibers in the organ of Corti. These results suggest possible roles of the 205 kDa MAP in the formation and maintenance of the highly polarized morphology of the epithelial cells of the organ of Corti, through stabilization and modulation of microtubule networks of these cells.


Changes of labile metabolites during anoxia in moderately hypo- and hyperthermic rats: correlation to membrane fluxes of K+.

The objective of this study was to assess the influence of temperature on the coupling among energy failure, depolarization, and ionic fluxes during anoxia. To that end, we induced anoxia by cardiac arrest in anesthetized rats maintained at a body temperature of either 34 degrees C or 40 degrees C, measured extracellular K+ concentration (K+e), and froze the neocortex through the exposed dura for measurements of phosphocreatine (PCr), creatine (Cr), ATP, ADP, and AMP, glucose, glycogen, pyruvate and lactate content after ischemic intervals of maximally 130 s. Free ADP (ADPf) concentrations were derived from the creatine kinase equilibrium. Hypothermia reduced the initial rate of rise in K+e, and delayed the terminal depolarization; however, both hypo- and hyperthermic animals showed massive loss of ion homeostasis at a K+e of 10-15 mM. The initial rate of rise in K+e did not correlate to changes in ATP, or ATP/ADPf ratio, suggesting that temperature changes per se may control the degree of activation of K+ conductances. The results clearly showed that, in both hyper- and hypothermic subjects, energy failure preceded the sudden activation of membrane conductances for ions. The results indicate that temperature primarily influences membrane permeability to ions like K+e (and Na+), and that cerebral energy state is secondarily affected. It is proposed that the higher rate of rise of K+e at high temperatures accelerates ATP hydrolysis primarily by enhancing metabolic rate in glial cells.


Localization of the central antinociceptive effects of diclofenac in the rat.

The ethacrynic acid-induced writhing response (WR) in the rat was studied after microinjections of diclofenac 0.1 ng-1 microgram/0.5 microliter (0.32 pmol to 3.2 nmol) into several brain regions involved in control of nociceptive behavior. The WR was inhibited after injections into the periaqueductal grey matter (PAG), ventromedial thalamus (VM), medial preoptic area (MPA) and the nucleus raphe magnus (NRM). Morphine 50 ng/0.5 microliter (0.16 nmol) was used as a positive control and vehicle injections were performed as negative reference. After diclofenac, there was a dose-dependent reduction of the WR with a threshold dose of approximately 1-10 ng in all brain areas studied except the nucleus reticularis paragigantocellularis interna (NRPGi). Naloxone 50 ng/0.5 microliter (0.15 nmol) administered into the same site 30 min after diclofenac injection, antagonized the diclofenac-induced inhibition of the WR almost completely in PAG and VM. Previous results demonstrate a central, naloxone-reversible component in the analgetic action of diclofenac. A qualitatively similar, centrally induced inhibition of the WR may be elicited after injections into PAG, VM and NRM. Thus, in addition to its peripheral mechanism of action, the non-steroidal anti-inflammatory drug, diclofenac, has a central mechanism of action which directly or indirectly involves a central opioid component.


Clorazepate synchronizes cultured rat C6 glioma in the early G1 phase of the cell cycle.

A water soluble benzodiazepine, clorazepate, has been used to establish the point of benzodiazepine proliferative arrest in the rat C6 glioma. Clorazepate inhibited C6 proliferation in a dose-dependent manner with an IC50 value of 280 microM, as judged by a nuclei counting procedure. Release of cells from a 48 h exposure to 350 microM clorazepate, at which over 70% of the cells were arrested, resulted in a synchronous entry into S phase 8-9 h later, as evidenced by a sharp increase in the incorporation of [3H]thymidine. This restriction point was demonstrated to be 2-3 h into the G1 phase by measuring the length of G1 in synchronized populations of C6 cells obtained by selection of mitotic figures from an asynchronous culture. Synchronous arrest of C6 by clorazepate required an exposure period of 24-36 h, approximately twice the doubling time of the cell line. A morphological study confirmed an early G1 point of proliferative arrest. Clorazepate synchronized cells exhibited a uniform morphology with the majority of cells assuming a configuration representative of anchorage-dependent cells in an early phase of attachment. The majority of cells were somewhat rounded and attached to the substratum by cytoplasmic 'skirts' with punctate structures which may represent focal adhesion points.


Neuropeptides and monoamines in the torus semicircularis of the carp (Cyprinus carpio).

The distribution of vasoactive intestinal polypeptide, gastrin-releasing peptide, gamma-melanocyte-stimulating hormone, alpha-neo-endorphin, angiotensin II, cholecystokinin-8, serotonin, and tyrosine hydroxylase has been studied in the nuclei lateralis and centralis of the Cyprinus carpio torus semicircularis using an indirect immunoperoxidase technique. In both nuclei, we found vasoactive intestinal polypeptide, gastrin-releasing peptide, gamma-melanocyte-stimulating hormone, alpha-neo-endorphin, serotonin, and tyrosine hydroxylase immunoreactive fibers, whereas the torus semicircularis was not immunoreactive for cholecystokinin-8 and angiotensin II. Moreover, no immunoreactive cell bodies containing peptides or monoamines were observed. The presence of these peptides and monoamines in both the nuclei lateralis and centralis suggests that such substances might be involved in the control of the visual, auditive, and/or lateral line information systems.


Binocularity and excitability loss in visual cortex cells of corpus callosum transected kittens and cats.

The contribution of the corpus callosum to binocularity of visual cortex cells and to their responsiveness was studied in cats. Electrophysiological recordings of the responses of single cells to visual stimulation was performed in the callosal projection zone, visual cortex area 17-18 boundary in callosotomized cats. Callosotomy was carried out by transection of the visual segment of the corpus callosum in 6-7-week-old kittens and in acute and chronic adult cats (postoperative recovery time: 11 days-39 months). While in our normal cats the common proportion of binocularly driven cells (79.8%) was found (66.3% in the sham controls), a remarkable diminution (29.7%) was found in the callosotomized kittens, in the acute (39.7%) and in the chronic (50.6%)-operated cats. We have also found a change in the amount of binocularity as function of postoperative recovery time. While the proportion of binocular cells was conceivable (60.7%) in the short- and intermediate-term callosotomized cats (postoperative time: 0.3-5.5 months), it was diminished (36.9%) in the long-term (6.5-39 months) chronic cats. As to the responsiveness level, it was found that visual responsive cells constituted 88% of the cells in the normal and 80.3% in the sham controls. In comparison, they constituted 69.2% in the acute, 54.4% in the chronic and 52.8% in the callosotomized kittens. Furthermore, callosal transection had produced a symmetric effect in the two hemispheres, regarding binocularity and responsiveness. It has been thus concluded that the corpus callosum is essential for the mediation of binocular functions between the two hemispheres; in addition, cortical excitability has been also found to depend on callosal integrity.


Protein kinase C activators stimulate beta-endorphin secretion from hypothalamic cells.

Relatively little is known about the regulation of secretion of hypothalamic beta-endorphin, the potent opioid that is believed to play a variety of physiological roles in brain. Previous work has shown that arginine vasopressin (AVP), which acts in brain primarily via activation of the phosphoinositol (PI) second messenger system, stimulates secretion of hypothalamic beta-endorphin. To test the hypothesis that activators of protein kinase C (PKC), which is activated following PI hydrolysis, stimulates secretion of beta-endorphins from hypothalamus, we studied the separate effects of stimulators of PKC including phorbol ester 12-myristate-13-acetate (PMA) and 1-oleolyl-2-acetyl glycerol (OAG- a diacyl glycerol analogue) on secretion of immunoreactive (IR-) beta-endorphin (measured by RIA) from dissociated fetal rat hypothalamic cell cultures. We also studied AVP and angiotensin II (Ang II), hypothalamic peptides which activate the PI second messenger pathway, and interactions of PMA and forskolin (FSK), an activator of the cyclic AMP/protein kinase A (PKA) pathway. PMA, OAG, AVP, and Ang II stimulated IR-beta-endorphin secretion. The stimulatory effect of both PMA and FSK on IR-beta-endorphin secretion was greater than that of PMA or FSK alone and was essentially additive.(ABSTRACT TRUNCATED AT 250 WORDS)


Tyrosine pretreatment alleviates suppression of schedule-controlled responding produced by corticotropin releasing factor (CRF) in rats.

Disruption of performance observed when animals are exposed to physical stressors which deplete brain catecholamines can be alleviated by pretreatment with the catecholamine precursor tyrosine. Central administration of the stress hormone corticotropin releasing factor (CRF) has been shown to affect a variety of behaviors and also to potently increase the release of central catecholamines. Since CRF-induced disruption of behavior may involve CRF-induced depletion of brain catecholamines, the present study examined whether tyrosine would alleviate suppression of schedule-controlled responding in rats resulting from ICV administration of CRF. Administration of CRF (1.0 microgram-10 micrograms) produced dose-dependent suppression of response rate and total number of earned reinforcers in rats responding on a multiple fixed-interval 60 s/fixed-ratio 20 schedule for food reinforcement. Pretreatment with 200 mg/kg tyrosine (IP) administered with ICV saline decreased response rate but did not lower total reinforcers, whereas 400 mg/kg of tyrosine decreased both. Injection of 400 mg/kg tyrosine reduced, but did not completely restore, CRF-induced suppression of behavior. The 200 mg/kg tyrosine dose was less effective in alleviating CRF-induced suppression of performance. These data indicate that pretreatment with the catecholamine precursor tyrosine can partially ameliorate performance decrements resulting from CRF administration.


Inhibitory inputs to the subfornical organ from the AV3V: involvement of GABA.

Action potentials were recorded extracellularly from single neurons in the subfornical organ (SFO) of the pentobarbital-anesthetized cat following stimulation of the regions surrounding the anteroventral third ventricle (AV3V). Of 328 SFO neurons studied, 103 were antidromically activated, showing direct projections from the SFO to the AV3V. However, the major effects of stimulations of the AV3V on SFO neurons were orthodromic inhibition; almost 30% of SFO neurons were inhibited by various sites in the AV3V, while a smaller proportion of cells were excited. Local application of bicuculline, an antagonist for GABA, attenuated the inhibitory responses induced by stimulation of the AV3V in seven out of eight neurons tested. Application of GABA inhibited 16 out of 24 neurons, while that of bicuculline alone excited 11 out of 26 neurons, suggesting the tonic inhibitory action of GABA on some SFO neurons. On the other hand, application of kynurenic acid, a nonspecific antagonist for the excitatory amino acids, did not affect the excitatory responses induced by stimulation of the AV3V, but kynurenic acid itself inhibited 6 out of 18 neurons tested. Application of glutamate excited most SFO neurons. This suggests that the excitatory amino acids may be the transmitter(s) of interneurons in the SFO but may not mediate the excitation from the AV3V.


Induction of c-fos immunostaining in the rat brain after the systemic administration of nicotine.

To search for evidence of altered neuronal gene expression in response to exposure to the highly addictive drug nicotine, rat brains were examined by immunocytochemistry for the fos protein after the systemic administration of nicotine. The drug was administered as an IV infusion over 1 h At a dose of 2 mg/kg, the most dramatic nicotine-induced fos nuclear immunostaining was seen in central visual pathways, including the superficial superior colliculus and the medial terminal nu. of the accessory optic tract, in the interpeduncular nu. Notably, many regions with high levels of nicotine binding sites, including the medial habenula, thalamus, substantia nigra, and ventral tegmental area, failed to express the c-fos gene with this schedule of nicotine administration. A minimal increase in fos immunostaining was seen after a nicotine dose of 0.5 mg/kg, with a much greater response after 1 or 2 mg/kg. The response was seen as soon as 60 min after the beginning of the infusion, was maximal at 2-3 h, and declined thereafter. c-fos expression was substantially attenuated in the superficial gray layer of superior colliculus, medial terminal nucleus of the accessory optic tract, and the interpeduncular nucleus by pretreatment with the centrally acting nicotine antagonist mecamylamine, 5 mg/kg IP, but not with the peripherally acting antagonist hexamethonium, 4 mg/kg IP. These observations identify a subset of central nervous system neurons that respond to nicotine with altered expression of the immediate early gene c-fos. These neurons presumably undergo long-term changes in gene expression as a result of acute exposure to high doses of nicotine.