Propranolol for the treatment of cyclosporine-induced headaches.

We studied whether small doses of propranolol given orally have an effect on headaches that are associated with intravenous cyclosporine therapy. In seven patients who had severe cephalalgia associated with intravenous cyclosporine post-bone marrow transplant, oral propranolol promptly relieved the symptoms in four patients. Intravenous propranolol was not effective in one patient who was unable to take oral medications. Propranolol should be considered as an alternative to chronic narcotics in patients with headaches due to cyclosporine.


The effects of body size on functional properties of middle ear systems of anuran amphibians.

Both tympanic and nontympanic pathways of sound reception are utilized by anuran amphibians. The relationship between body size and the acoustic responsiveness of various body surfaces that may serve as pathways for sound reception in anurans was analyzed. The motion of the different surfaces (tympanum, lateral body wall, lateral head surface, and dorsal shoulder surface) produced by sound was measured with a laser vibrometer in anuran species. The frequency response and amplitude levels of motion of these body surfaces clearly were linked with size. In all animals, nontympanic surfaces were most responsive to low frequencies, and the tympanum was most responsive to high frequencies. However, the responsiveness of nontympanic surfaces was greater, and extended to higher frequencies, in small anurans. In the smallest animals studied, nontympanic surfaces were often more responsive than the tympanum up to frequencies as high as 2500 Hz. In larger anurans, nontympanic responsiveness tended to decrease, and tympanic responsiveness tended to increase. In the largest animals studied, the tympanum was the most responsive surface at all except very low frequencies below about 200-300 Hz. These results suggest that small anurans can utilize nontympanic pathways for effective sound reception over a broad frequency range, whereas large anurans are more restricted to using a standard tympanic middle ear for hearing. This effect of body size on the utility of nontympanic sound reception may explain evolutionary patterns of tympanic ear reduction and loss observed in several small species of anurans.


Auditory cortex of the long-eared hedgehog (Hemiechinus auritus): II. Tuning properties.

The forebrain of hedgehogs is considered by many investigators as one of the simplest and most primitive among extant placental mammals. In a recent study we have shown that the auditory cortex of the long-eared hedgehog (Hemiechinus auritus) comprises two distinct auditory fields, which are tonotopically organized. In this study, we describe tuning properties of single cells in these two fields. Application of the Q10dB and Square Root measures for determining sharpness of tuning revealed that, although most of the cells in the more anterior field, which is considered primary, are sharply tuned, on the average they are more broadly tuned than cells in the primary auditory cortex of other mammals. In the posterior field, the distribution of narrowly and broadly tuned cells is equal. Narrowly tuned cells in both fields are equally narrow, as are the broadly tuned cells. Latencies of single cells in both fields are frequency and intensity dependent and are somewhat longer than these found in other mammals. The distribution of BFs vs. threshold intensity matches fairly well the behavioral audiogram previously described. Our findings suggest that, in spite of the view that the isocortex of hedgehogs represents a 'primitive' condition, some basic tuning properties of their auditory cortex cells are comparable to those of other mammals.


The visual forebrain and eating in pigeons (Columba livia).

The contribution of forebrain structures to the control of visually guided eating behaviors was studied using a technique for reversible 'visual decerebration'. The procedure is based upon the fact that structures in the thalamus and telencephalon receive their visual inputs primarily from the contralateral eye. When the eye contralateral to the ablated hemisphere is occluded, the remaining eye has unilateral access to these structures. When the eye ipsilateral to the ablated hemisphere is occluded, the bird is functionally decerebrate; i.e., visual processing by the remaining eye is restricted to structures caudal to the forebrain. The performance of normal and hemispherectomized subjects under binocular and monocular (unilateral, decerebrate) viewing conditions was compared on tests of ingestive efficiency, identification, conditioned peck localization, and grasping. In normal subjects, differences between right and left eye were not significant on any of these tasks. In hemispherectomized subjects, monocular performance on the first three tasks depended critically upon which eye was occluded. In the decerebrate condition (i.e., when the eye opposite the ablated hemisphere was used) performance on the identification, ingestive efficiency and peck localization tasks was significantly degraded, but grasping was unimpaired. We conclude that the brain structures critical for the visuomotor control of grasping lie caudal to the forebrain.


Retinal projections in the bowfin, Amia calva: cytoarchitectonic and experimental analysis.

The retinofugal projections in the bowfin, a non-teleost actinopterygian, were studied by autoradiographic and horseradish peroxidase methods, and the cytoarchitecture of retinorecipient regions of the diencephalon was analyzed with serially sectioned, Bodian stained material. Nuclei were identified in the thalamus, the periventricular portion of the posterior tuberculum, synencephalon, and pretectum which are homologous to like-named nuclei in teleosts and other non-teleost actinopterygian fishes. Of particular note, a posterior pretectal nucleus and, possibly, a homologue of nucleus corticalis were found to be present in the pretectum. These nuclei have previously been identified only in teleosts. The posterior pretectal nucleus is relatively small in the bowfin, and the distribution of a small, versus a large, posterior pretectal nucleus in Teleostei and Halecomorphi suggests that this nucleus was small plesiomorphically. The pattern of retinofugal projections in the bowfin is similar to that in other non-teleost actinopterygian fishes and in teleosts in most regards. Contralaterally, the retina projects to nuclei in the dorsal and ventral thalamus, superficial and central pretectum, dorsal and ventral accessory optic nuclei, and to the optic tectum. Additionally, there are sparse projections to the suprachiasmatic nucleus in the preoptic area, the periventricular nucleus of the posterior tuberculum, and the dorsal and ventral periventricular pretectal nuclei. Ipsilateral projections are sparse and are derived from fibers which do not decussate in the optic chiasm. Undecussated ipsilateral retinal projections, as present in the bowfin, are a widely distributed character in vertebrates and appear to be plesiomorphic for vertebrates.


Ketanserin reduces neuronal calcium accumulation and cell death in the hippocampus of the Mongolian gerbil after transient forebrain ischemia.

Excessive neuronal activity combined with an increased release of neurotransmitters is supposed to contribute to the delayed neuronal degeneration in animal models of transient cerebral ischemia. Since evidence is accumulating that serotonin (5-HT) exerts an excitatory effect on neurons via 5-HT2 receptors we tested the hypothesis that 5-HT2 receptor antagonists could protect neurons in the gerbil after transient bilateral carotid occlusion. In a first series of experiments, the 5-HT2 receptor antagonist ketanserin was injected intraperitoneally 15 min prior to 5 min of forebrain ischemia and given twice daily on the following 3 days. At a dose of 10 mg/kg i.p., the number of intact hippocampal CA1 neurons was significantly higher than in the saline-treated group and reached 74% of the sham-operated controls. In addition, the degree of neuronal damage correlated with an increased intracellular Ca2+ content in CA1 pyramidal neurons as revealed by arsenazo(III) staining with a procedure modified for paraffin sections. In a second series of experiments, ketanserin (10 mg/kg) was injected at various times after onset of ischemia. Up to a period of 90 min after ischemia, the number of intact CA1 pyramidal cells in ketanserin-treated animals was still significantly higher than in the saline-treated group. These results indicate that 5-HT2 receptor antagonists may protect neurons against ischemic damage even when the treatment is started after onset of ischemia. It remains to be investigated whether the neuroprotective effect of ketanserin is due to a neuronal action or to an inhibition of cerebrovascular vasospasm.


A novel hypothalamic-dispersed pituitary co-perifusion model for the study of growth hormone secretion.

This study presents a novel, in vitro, hypothalamic-dispersed pituitary co-perifusion system (HPPS) developed to examine the influence of the hypothalamus on pituitary growth hormone (GH) secretion in a controlled environment. In this perifusion system, dispersed rat pituitary cells were loaded onto Biogel P-2 (P-2) beads in a 0.5-ml plexiglas chamber and were submerged in a 37 degrees C water bath. After stabilization, two hypothalami were placed into each chamber on a thin layer of P-2 beads and the chamber was re-equilibrated. To test the system, pituitary cells were stimulated either directly with growth hormone-releasing factor (GRF) or indirectly via the hypothalamus, with clonidine, an alpha 2-adrenergic (alpha 2) receptor agonist. Perifusion of HPPS or pituitary cells with GRF (40 ng/ml) induced a substantial endogenous GH surge. Clonidine (2 x 10(-8) M) treatment stimulated a GH surge in HPPS chambers, but not in chambers containing only pituitary cells. Thus, somatotrophs respond to hypothalamic factors released in response to clonidine and not directly to alpha 2 stimulation. To determine if the components involved in GH feedback are present in the perifusion system, HPPS chambers were sequentially perifused with hGH, clonidine, and GRF. hGH pretreatment suppressed the clonidine but not the GRF-induced GH surge(s) observed in chambers perifused with clonidine and GRF only. In chambers only containing pituitary cells, GH was only increased in response to GRF when sequentially perifused with all three substances. This study demonstrates the dynamic interaction between the hypothalamus and pituitary in the regulation of GH secretion in vitro.(ABSTRACT TRUNCATED AT 250 WORDS)


Assessment of dopamine and its metabolites in the intracellular and extracellular compartments of the rat striatum after peripheral administration of L-[11C]dopa.

Success in the synthesis of L-3,4-[beta-11C]dihydroxyphenylalanine (L-[11C]DOPA) and its application to positron emission tomography encouraged us to perform radioactive metabolite analyses in rats in an early phase after peripheral injection of L-[11C]DOPA. Following intravenous injection of [11C]DOPA, the radioactivity associated with DOPA and its metabolites was determined in the striatum after decapitation and in striatal extracellular fluid using in vivo brain microdialysis. Without pretreatment, 70-80% of 11C-radioactivity taken up into the striatum was associated with acidic metabolites of dopamine (DA) from 2 to 30 min after administration of L-[11C]DOPA with or without 300 micrograms/kg of unlabelled L-DOPA. In contrast, 80-90% of 11C-radioactivity in the striatum was associated with DOPA and DA after pretreatment with benserazide (25 mg/kg, i.p.) followed by administration of L-[11C]DOPA with or without unlabelled L-DOPA. The radioactivity in the DOPA fraction decreased with time (from 35% of 11C-radioactivity in the striatum at 5 min to 10% at 30 min), but that in the DA fraction increased (from 57% to 68%). The 11C-radioactivity in the extracellular fluid determined by brain microdialysis was less than 0.4% of that in the whole striatum and no radioactivity was present in the DA fraction. These results suggest that, in an early phase after administration of L-[11C]DOPA, [11C]DA is the main metabolite and is localized exclusively in the intracellular compartment within this time frame.


Sex differences in estrogen receptor binding in the rat hypothalamus: effects of subsaturating pulses of estradiol.

The effects of subsaturating pulses of estradiol on cell nuclear retention of estrogen receptors in brain regions of male and female rats were determined. In the first experiment, age-matched adrenalectomized/gonadectomized (ADX/GDX) rats were killed 1 h after i.v. injection of a subsaturating dose of free estradiol (1.0 microgram/kg b. wt.) and cell nuclear estrogen receptor binding was measured in microdissected brain areas by an in vitro exchange assay. As we have previously reported with saturating doses of estradiol (3.6-36.0 micrograms/kg b. wt.), a greater concentration of receptors was measured in the ventromedial nucleus (VMN), periventricular preoptic area (PVPOA), and medial preoptic area (mPOA) of the female than in the same regions of the male brain. Binding levels did not differ between the sexes in the bed nucleus of the stria terminalis, arcuate nucleus, or the corticomedial amygdala. In the second experiment, GDX/ADX male and female rats received either a single or double pulse of 0.5 microgram estradiol/kg b. wt. administered 6 h apart. Rats were killed 1 h after the second pulse. In animals treated with a single pulse of estradiol 1 h before sacrifice, higher concentrations of estrogen binding were measured in the female VMN and PVPOA than in the male. Except in the mPOA, the concentration of estrogen receptor binding measured was not different from that measured in animals that had received an additional, earlier pulse of estradiol. In this area, receptor concentrations were lower in male rats treated with two pulses than in males treated with one pulse.(ABSTRACT TRUNCATED AT 250 WORDS)


Carbachol-induced rhythmic slow activity (theta) in cat hippocampal formation slices.

Application of the cholinergic agonist, carbachol, produced theta-like rhythmical waveforms, recorded in the stratum moleculare of the dentate gyrus in the cat hippocampal formation slices. This effect of carbachol was antagonized by atropine but not D-tubocurarine. These results provide first direct evidence that the hippocampal formation neuronal network in the cat is capable of producing synchronized slow wave activity when isolated from pulsed rhythmic inputs of the medial septum.


Induction of c-fos mRNA in rat brain by conditioned and unconditioned stressors.

Intense depolarizing stimuli induce the expression of the proto-oncogene c-fos which may be useful as a marker of neuronal activity. To determine if mild physical and behavioral stressors may also induce c-fos expression, we subjected rats to an unconditioned stressor (footshock) or a conditioned stressor (a tone previously paired with footshock) and measured c-fos mRNA levels in various brain regions using in situ hybridization. Removing rats from their home cage and exposing them to a tone was sufficient to cause increases in c-fos mRNA in several forebrain areas while further increases in c-fos occurred in the septum, cingulate cortex, and endopiriform nucleus in response to acute footshock stress. Both unconditioned and conditioned stressors increased c-fos mRNA levels in the locus ceruleus which correlated with stress-induced plasma corticosterone concentrations. Unconditioned footshock stress also increased c-fos mRNA in the hypothalamic paraventricular nucleus (PVN). However, neither conditioned nor unconditioned stressors induced c-fos in the PVN in rats which had been previously exposed to footshock. C-fos appears to be a sensitive marker for stress-responsive brain areas and may be important in mediating long-term neurochemical changes that result from stress.


The central effect of methionine-enkephalin on NK cell activity.

The central effect of opioid peptide on natural killer (NK) cell activity in BALB/c mice was investigated. Injection of methionine-enkephalin (Met-Enk), 0.02 microgram/mouse or 1 microgram/kg, directly into the cisterna magna (CM) of the brain, resulted in a significant enhancement of NK cell activity. This enhancement was blocked by opiate antagonists, naltrexone and quaternary naltrexone. The same dose of Met-Enk had no effect on NK cell activity when given to the mouse intraperitoneally or intravenously. Moreover, des-tyrosine-methionine-enkephalin injected into the CM at 1 microgram/kg, had no effect on NK cell activity. The results indicate that activation of an opioid-mediated pathway in the central nervous system is capable of activating the pathways that stimulate the NK cell response in the periphery.


Fast cyclic voltammetry of oxidizable species; depth profile in the rat striatum with and without potassium stimulation of the median forebrain bundle.

Fast cyclic voltammetry was used to monitor the release of dopamine in the caudate nucleus following potassium stimulation of the median forebrain bundle and to determine the characteristics of the small basal oxidation current while dipping of an electrode with a large tip (0.3 mm) in the caudate nucleus. The oxidation current was shown to be greater just after the electrode reached the caudate area than at a time 15-20 s after its arrival. There was less current increase observed when the tip reached the cortex and no current was observed in the white matter or with the electrode returning or with repeated passage along the same track. Potassium stimulation of median forebrain bundle induced an initial low magnitude dopamine release which disappeared within minutes. The basal oxidation current increased step by step after a few potassium stimulations. This effect was not blocked by pargyline (75 mg/kg) whereas L-DOPA (200 mg/kg) was shown to increase it. It is thought that the large tip of the lowered electrode can destroy neurons and induce the release of intracellularly stored compounds into the extracellular space. We consider also dopamine to be responsible for the increase in oxidizable current just after the electrode movement stopped.


Behavior of inhibitory and excitatory propriobulbar respiratory neurons during fictive vomiting.

The behavior of propriobulbar respiratory neurons was studied during fictive vomiting in decerebrate, paralyzed, artificially ventilated cats. Fictive vomiting was identified by a characteristic series of synchronous phrenic and abdominal nerve bursts, induced by electrical stimulation of abdominal vagal afferents and/or i.v. infusion of emetic drugs. Data were obtained from inspiratory neurons having decrementing (I-DEC) or constant (I-CON) discharge patterns and expiratory decrementing (E-DEC) neurons located in the Bötzinger complex and adjacent rostral ventral respiratory group. These neurons are known to make excitatory (I-CON) and inhibitory (I-DEC, E-DEC) connections with a variety of medullary respiratory neurons. During fictive vomiting: 8 of 14 I-DEC neurons fired in phase with synchronous bursts of phrenic and abdominal nerve discharge; the other 6 were silent. Of 12 I-CON neurons, 5 fired in phase with phrenic and abdominal bursts; 7 were silent. All (6) E-DEC neurons were either silent or fired weakly between bursts of phrenic and abdominal discharges. The possible roles of I-DEC and I-CON neurons in actively reorganizing the behavior of other respiratory neurons during fictive vomiting are discussed. In particular, the firing of many I-DEC neurons was found to be appropriate to inhibit inspiratory, and two types of expiratory, bulbospinal neurons during fictive vomiting.


Fos-like immunoreactivity in the rat superficial dorsal horn induced by formalin injection in the forepaw: effects of dorsal rhizotomies.

As previously described at the lumbar spinal level, we found that 2 h after subcutaneous formalin injection in the distal part of the fore-limb, Fos-like immunoreactivity (FLI) was induced in the ipsilateral cervical enlargement. Not surprisingly, as the injection site corresponds to the distal part of the C6-C8 dorsal root dermatomes, maximal labelling which predominated in the superficial laminae, was observed in the C6-C8 segments and to a lesser extent in C5. Similar experiments were performed on rats which underwent various types of unilateral dorsal rhizotomies (DRh) 7 days before formalin injection. In animals with C4, C5, T1 and T2 DRh sparing C6-C8 the rostrocaudal distribution was similar to the intact one. But, in animals having C4-T2 DRh sparing one single root, C7, the segmental FLI distribution was modified: it was slightly increased in C7, decreased in C6 and significantly decreased in C8. As expected, no FLI was found in animals with C4 to T2 DRh. The spared root model provides information about the segmental distribution in the cervical spinal cord of the input brought by a single root following stimulation of the distal forelimb, i.e., maximal distribution in the entry segment, but also in the two rostral and one caudal segments.


The neurotoxic effects of p-chloroamphetamine in rat brain are blocked by prior depletion of serotonin.

Systemic administration of p-chloroamphetamine (PCA) causes degeneration of serotonergic (5-HT) axons, but recent data indicate that this drug itself is not neurotoxic when applied directly to 5-HT axons. The present study was designed to test whether the toxic effects of PCA in the brain are dependent on release of endogenous 5-HT and to identify which stores of 5-HT are involved. The long-term effects of PCA on brain levels of 5-HT and on central 5-HT axons were determined in rats that had been initially depleted of 5-HT by administration of p-chlorophenylalanine and reserpine. The results show that transient depletion of 5-HT provides substantial protection against subsequent PCA-induced degeneration of 5-HT axon terminals; the neurotoxicity induced by PCA thus appears to be dependent on the presence of endogenous stores of 5-HT. In addition, the protective effect of 5-HT depletion is found only after pretreatment regimens that deplete peripheral as well as central stores of 5-HT. We interpret this finding as evidence that release of 5-HT from peripheral storage sites may be necessary for the expression of PCA-induced toxicity. Based on these results, we propose that central neurotoxicity is not induced by a direct action of PCA alone but may require or be augmented by a toxic metabolite of 5-HT.


Bradykinin modulation of a spinal nociceptive reflex in the rat.

Bradykinin (BK) is a potent algesic compound. Therefore, we hypothesized that BK, acting as a peripheral noxious stimulus, would attenuate or inhibit responses to another noxious stimulus. When administered intravenously (i.v.) to rats lightly anesthesized with pentobarbital, BK produced a dose-dependent (12-144 micrograms/kg) inhibition of the nociceptive tail-flick (TF) reflex. BK also produced a dose-dependent decrease in mean arterial blood pressure, a subsequent increase in heart rate and an increase in the rate of respiration. The latency to the maximal effect of BK on the TF reflex was 10 s and was occasionally preceded by pseudoaffective responses at doses greater than 48 micrograms/kg BK. Neonatal treatment with capsaicin (50 mg/kg, subcutaneous) significantly attenuated BK-produced inhibition of the TF reflex indicating that BK was acting via peripheral afferents to inhibit the TF reflex. Reversible cold block or complete spinal transection at a low thoracic level (T9-12), but not reversible cold block at a high cervical level (C1-2), significantly attenuated BK-produced inhibition of the TF reflex, suggesting that BK activates afferents which enter the spinal cord between C2 and T9. Pretreatment with intrathecally administered phentolamine (30 micrograms), an alpha-adrenoceptor antagonist, or methysergide (30 micrograms), a non-selective serotonin receptor antagonist, did not alter BK-produced inhibition of the TF reflex, further supporting the absence of activation of descending systems from the brainstem by i.v. BK. While coadministration of PGE2 and BK significantly potentiated BK-produced inhibition of the TF reflex, neither bilateral removal of the stellate ganglion nor bilateral cervical vagotomy significantly affected the inhibitory action of i.v. BK on the TF reflex. These results suggest that i.v. BK inhibits the nociceptive TF reflex by activation of capsaicin-sensitive visceral afferents entering the spinal cord between C2 and T9.


Behavioral modulation of sensory responses of primate putamen neurons.

Sensory response properties of putamen neurons were investigated in 4 awake macaque monkeys while the animals performed GO/NO-GO tasks guided by sensory stimuli. In the GO condition, visual stimuli served as a trigger for a sequence of repetitive elbow flexion-extensions and auditory stimuli served as a trigger for orofacial movements, while the identical sensory stimuli required the animals to withhold movements in the NO-GO condition. Short latency responses to visual and/or auditory stimuli were observed in 206 type I cells with tonic discharges at about 2-7 Hz, with a characteristic discharge rhythm and in 57 type II cells with low spontaneous discharge rates, usually less than 0.5 Hz, which showed burst discharges during the tasks. Most of these cells (163/206 type I cells, 52/57 type II cells) responded differentially to identical sensory stimuli presented in the GO and NO-GO conditions. The responses to sensory stimuli presented in the GO condition were much larger in 67% of type I cells and 58% of type II cells than the responses to identical stimuli presented in the NO-GO condition. On the other hand, 12% of type I and 33% of type II cells showed larger responses in the NO-GO than in the GO condition. It is suggested that the strong behavioral modulation of striatal sensory responses serves as a mechanism to start the neural processes for preparation and/or initiation or withholding of particular movements associated with learned sensory stimuli.


Distribution of [3H]clonidine binding sites in the brain of the convulsive mutant quaking mouse: a radioautographic analysis.

The radioautographic analysis of [3H]clonidine binding was performed on brain slices from the convulsive mutant mice quaking and their controls of the same strain. In the quaking mice significant increases were observed mostly in the brainstem and the cerebellum, but also in a few regions of the forebrain, such as the lateral and medial thalamic nuclei, the medial geniculate nucleus, the amygdala and the hypothalamus. Other regions, such as the cerebral cortex and the hippocampus, which are classically involved in various models of epilepsy, but not in the quaking mice, did not show any modification of [3H]clonidine binding. A high degree of correlation was found between the structures with an increased density of alpha 2-adrenoceptor binding sites and the distribution of regions from which seizures can be elicited by intracerebral electrical stimulation in head-restrained quaking mice. This comparison emphasizes the role of noradrenaline acting at the level of alpha 2-adrenoceptors in the epileptic syndrome of the quaking mutants.


Carnosine-like immunoreactivity is associated with synaptic vesicles in photoreceptors of the frog retina.

The distribution of carnosine-like immunoreactivity in photoreceptors of the frog retina was studied by post-embedding electron microscope immunocytochemistry. Different fixation and embedding procedures were applied and the best results were achieved on sections from tissue embedded in the hydrophilic resin LR White. All photoreceptor types present in the frog retina (red and green rods, single and double cones) were intensely reactive for the carnosine antiserum. The immunoreactivity was particularly prominent in the synaptic terminal, nucleus and myoid, but decreased in the ellipsoid and in the outer segment. Müller glial cells and horizontal and bipolar cell processes in the outer plexiform layer were completely unstained. At the level of the photoreceptor terminals, the presence of gold particles within the synaptic vesicles could be demonstrated. This study is the first to provide the anatomical evidence for the presence of carnosine in the synaptic vesicles of a nervous cell. The present data may be relevant to the understanding of the biological functions of carnosine (and related dipeptides) and offer an exploitable system in which to verify its proposed role as a neurotransmitter or neuromodulator in sensory systems.


Semi-quantitative analysis of the effects of estrogen on CGRP- and methionine-enkephalin-immunoreactivity in the periventricular preoptic nucleus and the medial preoptic area of female rats.

The effects of 17 beta-estradiol (E2) on calcitonin gene-related peptide (CGRP)- and methionine-enkephalin (Met-Enk)-immunoreactive (IR) nerve fibers in the periventricular preoptic nucleus (Pe) and the medial preoptic area, including the medial preoptic nucleus (MPN), of the female rat were studied semi-quantitatively by using a computer-based image analysis system. The area occupied by CGRP- and Met-Enk-IR fibers was significantly increased in the Pe and the MPN after 28-day exposure to E2. Computer-based analysis of CGRP- and Met-Enk-IR fibers in an area of 50-microns intervals from the wall of the third ventricle showed a low flat histogram pattern in ovariectomized rats, but E2 treatment caused a diphasic pattern, corresponding to the Pe and the MPN, respectively. Since the Pe and the MPN contain a high population of estrogen receptors, it is suggested that E2 may have an influence on the neuronal configuration of afferent fibers to these areas.


Properties of favored patterns in spontaneous spike trains and responses of favored patterns to electroacupuncture in evoked trains.

By using 'the modified detection method', our previous study has shown that all spontaneous spike trains recorded from several areas of brain and spinal cord have favored patterns (FPs). The present study further shows that: (1) all newly detected spike trains from substantia nigra zona compacta, nucleus periventricularis hypothalami and nucleus hypothalamicus posterior also have FPs, and some spike trains from neurons in the same nucleus have a common favored pattern (CF, i.e. they share the same FP), indicating that FP and CF in spike trains are common phenomena; (2) all serial correlation coefficients of FP repetitions (in serial order) in different spike trains detected are less than 0.3 (close to 0), revealing that the repetition of FPs is a renewal process; (3) in different periods of the spike trains evoked by electroacupuncture (EA), the number of different FPs and the number of repetitions of the same representative FP either increase or decrease along with the change of firing rate. The tendencies of these changes are very similar, but after EA the repetitions of different FPs in the same spike trains change differently, showing that different (hidden) responses exist at the same time. The above results suggest that the FPs in spike trains may represent various neural codes, and 'the modified detection method of FP' can pick up more information from spike trains than the firing rate analysis, hence it is a very useful tool for the study of neural coding.


Nerve growth factor increases calcium binding protein (calbindin-D28K) in rat olfactory bulb.

Calbindin-D28K (CaBP28K) is a soluble intracellular protein capable of sequestering micromolar concentrations of calcium. The in vivo regulation of CaBP28K by recombinant human nerve growth factor (rhNGF) was studied in adult, male rats. Via Alzet 2002 pumps, each rat received, for 14 days, a lateral ventricle infusion (i.c.v.; n = 5-6/group) of 12 microliters PBS/day containing 1.0 microgram cytochrome C (control) or an equal amount of rhNGF. Six other animals received a vehicle or rhNGF infusion into the central neostriatum. CaBP28K was elevated by 75% (P less than 0.01) in the olfactory bulb following i.c.v. rhNGF in each of two experiments and was not altered in the temporal cortex, hippocampus, olfactory tubercle, cerebellum, or neostriatum. Direct striatal injections of rhNGF did not alter CaBP28K in the neostriatum or other regions (including the olfactory bulb). The increases in olfactory bulb CaBP28K protein levels were verified via Western blot analysis. CaBP28K immunocytochemistry revealed that 33% of olfactory bulb neurons are immunoreactive for CaBP28K and that the number or proportion of immunoreactive neurons did not change with i.c.v. infusions of rhNGF, suggesting that exogenously delivered rhNGF augments the content of CaBP28K in olfactory bulb neurons that normally express the protein. Endogenous NGF may function as a neuroprotective factor by enhancing the ability of these cells to sequester cytoplasmic calcium and retard calcium-mediated neurodegeneration.


Thermogenic actions of tryptophan in the rat are mediated independently of 5-HT.

Serotonin (5-HT) has been implicated in the central control of energy balance, via inhibition of food intake and stimulation of thermogenesis. Its rate of synthesis in brain is dependent on the availability of its precursor amino acid, tryptophan. The objective of the present study was therefore to investigate the thermogenic actions of tryptophan and to determine whether these actions are mediated by 5-HT. Central or peripheral injections of 5-HT (i.c.v.; 0.5-40 micrograms), 5-hydroxytryptophan (5-HTP) (i.c.v.; 20 micrograms) or tryptophan (i.p.; 20 mg/kg, i.c.v.; 12-60 micrograms) significantly increased resting oxygen consumption (VO2 by approximately 15-20%) in conscious rats, without apparent effects on physical activity. Small increases (5-7%) in VO2 were also observed following peripheral injections of aspartate or glycine (20 mg/kg) but not taurine, whilst central injections of tyrosine or leucine (15-18 micrograms) significantly increased VO2 by 15%. We have previously reported that the thermogenic and anorexic actions of 5-HT are mediated by corticotropin-releasing factor (CRF). In the present study, the thermogenic actions of 5-HTP, like those of 5-HT, were significantly reduced by pretreatment (5 min before) with the CRF antagonist alpha-helical CRF9-41 (25 micrograms, i.c.v.) or a polyclonal antibody to CRF. However, the thermogenic actions of tryptophan were not significantly modified by pretreatment with either the 5-HT antagonist, methysergide (20 micrograms, i.c.v.) or with the CRF antagonist or antibody and thus appear to act through different mechanisms to 5-HT.(ABSTRACT TRUNCATED AT 250 WORDS)


Differences in the number of arginine-vasopressin-immunoreactive neurons exist in the suprachiasmatic nuclei of house mice selected for differences in nest-building behavior.

Arginine-vasopressin (AVP) is a homeostatic modulator of body temperature during fever and may also be involved in normal body temperature control. In the present study the hypothalamus of mice bi-directionally selected for thermoregulatory nest-building behavior was immunocytochemically labeled for AVP. The low-selected mice had a 1.5-fold higher number of AVP-immunoreactive neurons in the suprachiasmatic nuclei (SCN) compared to the unselected control and the high-selected mice. No differences between the selected lines could be detected in the number of AVP-immunoreactive neurons in the paraventricular nuclei (PVN). The neuroanatomical data suggest a possible role of AVP in the SCN and control of thermoregulatory nest-building behavior. Our selected mice may prove to be a model system to study the role of AVP in the SCN.


Microdialysis perfusion increases the sensitivity of rat striatal neurons to ischemic insult.

Brief periods of global forebrain ischemia which never produced striatal cell loss in control, non-dialyzed striatum resulted in significant cell loss in the contralateral, dialyzed striatum. No striatal cell loss was seen following these short periods of ischemia in animals implanted with dialysis probes that were not perfused. Dialysis perfusion alters striatal extracellular fluid composition in such a way as to render this tissue more sensitive to brief ischemic insult.


Regulation of the prefrontal cortical dopamine system by the neuroactive steroid 3a,21-dihydroxy-5a-pregnane-20-one.

The dopamine innervation of the prefrontal cortex can be differentiated from other telencephalic dopamine projection fields by its sensitivity to stress. The stress-induced activation of the mesoprefrontal cortical dopamine system can be blocked by pretreatment with benzodiazepines. A group of neuroactive steroids that modulate GABA-induced chloride flux through means distinct from that of the benzodiazepines has recently been identified. Intraventricular administration of the neuroactive steroid 3 alpha,21-dihydroxy-5 alpha-pregnane-20-one resulted in a dose-dependent decrease in dopamine metabolites in the prefrontal cortex, but not in mesolimbic or striatal sites; sedative effects were not observed. Moreover, the neuroactive steroid selectively attenuated the stress-induced activation of the mesoprefrontal cortical dopamine system. These data suggest that neuroactive steroids may function as endogenous anxiolytic agents.


Regulation of intracellular free calcium in normal and dystrophic mouse cerebellar neurons.

We measured free intracellular calcium ([Ca2+]i) in cultured cerebellar granule cells from normal and mdx mice. Resting levels of ([Ca2+]i) were 24% higher in the dystrophic neurons (normal: 61.2 +/- 1.5 nM calcium, n = 104; dystrophic: 76.1 +/- 2.4 nM calcium, n = 136, P less than 0.01). Dystrophic neurons showed a significantly greater increase in ([Ca2+]i) in the presence of elevated (18 mM) extracellular calcium levels. Resting sodium levels ([Na+]i), however, were found to be similar in normal and dystrophic granule neurons. In addition, sodium influx rates after ouabain inhibition of the Na+/K+ ATPase were also identical. Therefore, the increased permeability of granule neurons was specific to calcium, and did not result from a non-selective cation-permeable conductance. Unlike granule cells, astrocytes do not express dystrophin. Glial cells from normal and dystrophic mice showed no difference in their resting free calcium levels or their response to a high calcium load. Thus, cerebellar granule neurons from mdx mice show a calcium-specific regulatory defect similar to that found in dystrophic muscle fibers, while cerebellar glial cells, which do not normally express dystrophin, have no such defect.


Antiestrogen binding sites in brain and pituitary of ovariectomized rats.

In addition to interacting with estrogen binding sites in a number of tissues, antiestrogens have recently been shown to interact with a separate, estrogen-non-compatible, antiestrogen-specific binding site (AEBS). In order to better understand possible mechanisms by which the antiestrogens may effect behavioral and physiological changes, we have examined AEBS in several areas of the brain and pituitary in adult, ovariectomized rats. Single point binding assays with 2 nM [3H]tamoxifen (TAM) in the presence of saturating amounts (1 microM) E and +/- 1 microM TAM indicated the existence of specific binding to AEBS throughout the brain and pituitary. In most areas of the brain (cortex, cerebellum, amygdala, area postrema/nucleus of the solitary tract region) as well as pituitary. Scatchard analyses revealed the presence of a single AEBS with a dissociation constant (Kd = 1-4 x 10(-9) M) similar to that previously reported for other tissues. However, in both hypothalamus and preoptic area, an additional, higher affinity site (Kd = 6-9 x 10(-11) M) was found. Competitive inhibition studies revealed that there was little competition by the potent estrogen agonist, diethylstilbesterol, for AEBS binding. Antiestrogens competed in the following order: tamoxifen greater than or equal to nafoxidine much greater than keoxifene. Additional competitive inhibition studies were run using neurotransmitter antagonists. The phenothiazines, chlorpromazine and fluphenazine, bind to both D1 and D2 dopamine receptors and effectively compete with [3H]TAM for binding at the AEBS. Other pharmacological substances, including specific antagonists of the D2 sites, as well as antagonists of the norepinephrine, opiate, histamine, GABA and acetylcholine systems, were ineffective competitors for [3H]TAM binding.


Regional localization of the regulatory subunit (RII beta) of the type II cAMP-dependent protein kinase in human brain.

The distribution of the regulatory (RII beta) subunits of type II cAMP-dependent protein kinase in cortical and subcortical areas was examined in human control and Alzheimer's disease (AD) brains. Four monoclonal antibodies generated against bovine brain RII, which cross-reacted with human brain RII beta, detected RII-immunoreactivity in pyramidal neurons of the hippocampus and frontal, occipital, parietal and superior temporal cortices and in non-pyramidal neurons of the amygdala and putamen. RII beta immunoreactivity was localized to neuronal perikarya, proximal dendrites and cell processes. With the exception of rare processes in the ventroposterior lateral nucleus, RII-immunoreactivity was not seen in the thalamus. Other areas lacking RII-immunoreactivity included the midbrain, caudate nucleus and globus pallidus. RII-immunoreactivity was not detected in endothelia or glia. Except for the neocortex, the distribution of RII beta immunoreactivity was the same in AD and non-demented control brains; however, cell bodies and their processes stained more intensely and uniformly in the neocortical regions of non-demented controls compared to AD. In the neocortex of AD, RII beta immunoreactivity was substantially decreased in the superior temporal and occipital cortices, but not in the frontal cortex. Our data suggest that RII subunits are regionally distributed in the human brain. RII-immunoreactivity was decreased in some regions of neocortex in AD, but it did not preferentially colocalize with neurofibrillary tangles (NFT), senile plaques, or neuropil threads.


Prenatal flutamide alters sexually dimorphic nuclei in the spinal cord of male rats.

The effects of prenatal exposure to the antiandrogen flutamide on two sexually dimorphic nuclei of the lumbar spinal cord, the dorsolateral nucleus (DLN) and the spinal nucleus of the bulbocavernosus (SNB), were investigated. Rat dams were given daily injections of 5 mg flutamide or vehicle alone from day 11 through 21 of pregnancy. The spinal cords and perineal morphology of their male and female offspring were examined in adulthood. Flutamide reduced the number of SNB and DLN neurons, reduced the somal and nuclear area of SNB neurons, and reduced the weight of the perineal muscles in males. Flutamide produced no effect in females. No sexual dimorphism was found in the mean somal area of DLN neurons, but a sexual dimorphism was found in the distribution of somal areas in our samples; females had proportionately more large neurons than males. Flutamide-treated males also had proportionately more large neurons than control males but fewer than females. A sexual dimorphism was found in the nuclear areas of DLN neurons but flutamide did not influence this trait.


Structural modeling of functional visual pathways mapped with 2-deoxyglucose: effects of patterned light and footshock.

This paper describes the first application of structural modeling to the visual system. Structural modeling, or path analysis, is a mathematical method that allows for the quantification of the functional strengths of anatomical connections between the structures that form a neural system. The objective was to demonstrate how structural modeling can be used to determine the functional interrelationships between brain structures that form the visual system and how these interrelationships change under different conditions. Data were obtained from measures of 2-deoxyglucose uptake in the visual system of rats presented with either patterned light or darkness. The effects of arousing footshock on visual system operations were also investigated. Models based on the anatomical connections and the interregional correlations between metabolic activity data were used to determine path coefficients representing the magnitude of the influence of each directional path. Statistical evaluation of the models revealed that the dominant positive influences on visual system activity in the darkness were the tectocortical subsystem and the descending connections from secondary visual cortex. In the patterned light model, the total influence of the geniculocortical subsystem was higher than in the dark, and the tectocortical pathways showed both a reduction and a shift in the direction of effects. The models also revealed that the effects of footshock-induced arousal on visual system operations depended upon the visual environment and on extra-visual influences. The footshock led to an increase in the interaction of the two main subsystems at the level of connections between primary visual cortex and the lateral posterior nucleus, and a descending negative influence from the secondary visual cortex became dominant. The models are discussed in the context of conventional analyses to show how structural modeling allows for the determination of much more information about the functional interactions within the visual system of subjects under different experimental conditions.


Evidence for a non-genomic action of progestins on sexual receptivity in hamster ventral tegmental area but not hypothalamus.

Progestogenic stimulation of both the ventromedial nucleus of the hypothalamus (VMH) and the ventral tegmental area (VTA) is critical for normal receptivity in estrogen-primed hamsters. However, anatomical and biochemical studies have identified very few estrogen-induced progestin receptors in the rodent ventral midbrain. To determine whether progesterone might be working on the membrane of neurons in the VTA, progesterone 3-CMO BSA (P-3-BSA) was applied intracranially. The size of P-3-BSA makes it relatively impermeable to the cell membrane. Ovariectomized hamsters were implanted with 2 chronic cannulae, one aimed at the VMH and the other at the contralateral VTA. These animals were then estrogen-primed and tested for sexual receptivity after progesterone-containing tubes were inserted just dorsal to the VMH and P-3-BSA inserts were applied above the VTA. The following week, the hamsters were tested again with the contents of the inserts reversed. Animals with progestogenic stimulation to the VMH and P-3-BSA to the VTA were receptive yet those with P-3-BSA to the hypothalamus and progesterone to the VTA were not receptive. These data suggest that progesterone is capable of facilitating sexual receptivity within the VTA by actions on the cell membrane. The non-genomic effects in the VTA require concurrent genomic activation by progesterone within the hypothalamus.


Short-day stimulation of testicular activity and immunoreactivity of the hypothalamic GnRH system in mink following deafferentation of the pineal body by bilateral superior cervical ganglionectomy and melatonin replacement.

The effects of superior cervical ganglionectomy on testicular function (testis volume and plasma testosterone levels) and the immunocytochemical activity of the GnRH hypothalamic system were studied in the mink, a short-day breeder. Animals reared in a natural photoperiod were (i) ganglionectomized at four different times during the period extending from the end of summer to the end of autumn (September 15, October 20, October 28, and December 1), and (ii) reared for 50 days in a short gonadostimulatory photoperiod (4L:20D). Lastly, an attempt was made to overcome the effects of superior cervical ganglion removal by administering melatonin to mink reared in a natural photoperiod. In mink reared in a natural photoperiod, deafferentation of the pineal on September 15 (L = 12.5 h) or October 20 (L = 10.5 h) resulted in consistently low values of testicular volume and plasma testosterone until the end of the experiment (February). When the operation was performed on October 28 (L = 10 h) testicular activity was initiated but only lasted a short time and did not allow maximal gonadal development. When superior cervical ganglionectomy was carried out on December 1 (L = 8.5 h), during the phase of renewed testicular activity, the increases in testicular volume and testosterone levels were not affected by the operation and the subsequent variation of these parameters was identical to that observed in intact animals. Similarly, in mink reared for 50 days in a photoperiod of 4L:20D before superior cervical ganglionectomy, deafferentation of the pineal did not prevent gonadostimulation induced by short days.(ABSTRACT TRUNCATED AT 250 WORDS)


Ibotenic acid lesion of the ventral hippocampus differentially affects dopamine and its metabolites in the nucleus accumbens and prefrontal cortex in the rat.

To determine the influence of neurons of the ventral hippocampus on dopamine (DA) turnover in other limbic areas, spontaneous and amphetamine-induced locomotion as well as DA and its metabolites were assayed in nucleus accumbens, medial prefrontal cortex and anteromedial striatum, 14 and 28 days after bilateral ibotenic acid (IA) or sham lesions of the ventral hippocampus in the rat. Spontaneous locomotion was increased 28 days postoperatively, while D-amphetamine induced locomotion was augmented both 14 and 28 days postoperatively in IA lesioned animals. DA levels in the nucleus accumbens were decreased on the 14th, but increased on the 28th day after the lesion. Dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA) and the DOPAC/DA ratio in the medial prefrontal cortex (MPFC) were reduced 28 days postoperatively. Moreover, there was a significant negative correlation between the DOPAC/DA ratio in the MPFC and DA levels in the nucleus accumbens at this time point. These data indicate that a lesion of the ventral hippocampus can produce differential changes in cortical and limbic DA activity. Implications for an animal model of schizophrenia are considered.


Spontaneous recovery of MPTP-damaged catecholamine systems in goldfish brain areas.

In goldfish, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) administered for 3 consecutive days (10 mg/kg), produced a marked decrease in dopamine (DA) and noradrenaline (NA) levels in telencephalon, diencephalon and medulla oblongata, without affecting the serotonin (5-HT) content. Furthermore the neurotoxin decreased either [3H]DA high affinity uptake or K(+)-stimulated DA release from synaptosomal (P2) preparations, with concomitant up-regulation of D2 postsynaptic receptors as well. No significant changes of choline acetyltransferase and glutamic acid decarboxylase activity or [3H]glutamate uptake were observed. Moreover the pretreatment with deprenyl (1 mg/kg) or mazindol (10 mg/kg) but not with clorgyline (5 mg/kg) prevented catecholamine depletion. Added in vitro to synaptosomal preparations both MPTP and more potently MPP+, in a concentration-dependent manner, inhibited [3H]DA uptake. Time course study revealed that MPTP-induced alteration of neurochemical parameters in goldfish brain areas were almost completely reversed within 6 weeks, suggesting that catecholamine systems in goldfish brain show a remarkable power of recovery after MPTP lesion.


Development of MEKA (phosducin), G beta, G gamma and S-antigen in the rat pineal gland and retina.

Pinealocytes and retinal photoreceptor cells contain an unusual cytoplasmic complex composed of the G beta gamma dimer of GTP-binding regulatory proteins (G-proteins) tightly bound to an acidic 33 kDa phosphoprotein termed MEKA or phosducin; MEKA is a substrate of cyclic AMP-dependent protein kinase. This study characterized the developmental appearance of these and two related proteins, G gamma and S-antigen, in pineal and retinal tissue. MEKA was absent in the pineal gland prior to birth, at a time when it was possible to detect G beta in pineal cytoplasm, indicating that the appearance of G beta in the cytoplasm precedes that of MEKA and does not appear to require the presence of MEKA. The absence of MEKA at this time indicates that the cyclic AMP stimulation of pineal serotonin N-acetyltransferase activity is not mediated by MEKA, which has been considered as a possible role of MEKA. After postnatal day 7, pineal MEKA and cytoplasmic G beta increased in a parallel manner, with peak values occurring at about postnatal day 21. Thereafter, both proteins in the pineal gland decreased in a parallel fashion to 10 and 35% of their peak values, respectively; in contrast, the cytoplasmic protein S-antigen and membrane associated G beta remained at maximal levels after this time. Whereas both MEKA and G beta decreased late in development in the pineal gland, these proteins either increased or remained constant in the retina. These tissue-specific patterns were found to differ from those of another cytosolic protein found exclusively in the pineal gland and retina, S-antigen, which remained constant after day 21 in the pineal gland but decreased in the retina late in life.


Dioctanoylglycerol stimulates accumulation of [methyl-14C]choline and its incorporation into acetylcholine and phosphatidylcholine in a human cholinergic neuroblastoma cell line.

Dioctanoylglycerol, a synthetic diacylglycerol, stimulated [14C]choline uptake in cultured human neuroblastoma (LA-N-2) cells. As this effect has not, to our knowledge, been reported before, it was of interest to characterize it in more detail. In the presence of 500 microM dioctanoylglycerol the levels of [14C]choline attained during a 2 hour labeling period were elevated by 78 +/- 12%, while [14C]acetylcholine and long fatty acyl chain [14C]phosphatidylcholine levels increased by 26 +/- 2% and 19 +/- 5%, respectively (mean +/- S.E.M.). Total (long chain plus dioctanoyl-) [14C]phosphatidylcholine was increased by 198 +/- 33%. Kinetic analysis showed that dioctanoylglycerol reduced the apparent Km for choline uptake to 56 +/- 9% of control (n = 4). The Vmax was not significantly altered. The stimulation of [14C]choline accumulation by dioctanoylglycerol was not dependent on protein kinase C activation; the effect was not mimicked by phorbol ester or by 1-oleoyl-2-acetylglycerol, and was not inhibited by the protein kinase C inhibitors H-7 or staurosporine, or by prolonged pretreatment with phorbol 12-myristate 13-acetate. The effect of dioctanoylglycerol was slightly (but not significantly) reduced by EGTA and strongly inhibited by the cell-permeant calcium chelator bis(o-aminophenoxy)-ethane-N,N,N',N'-tetraacetic acid, tetra(acetoxymethyl)ester. Although these results implicate elevated intracellular calcium in the response, dioctanoylglycerol did not increase phosphatidylinositol hydrolysis in LA-N-2 cells, and its effect was not inhibited by the diacylglycerol kinase inhibitor R 59 022 (which blocks the conversion of diacylglycerol to phosphatidic acid, a known stimulator of phosphatidylinositol hydrolysis).(ABSTRACT TRUNCATED AT 250 WORDS)


Intracerebroventricular application of competitive and non-competitive NMDA antagonists induce similar effects upon rat hippocampal electroencephalogram and local cerebral glucose utilization.

In this study we have used electrophysiological and metabolic markers to investigate the effects of competitive and non-competitive NMDA antagonists in rats after central or peripheral administration. The non-competitive antagonist, MK-801, induced dose-dependent suppression of rat hippocampal EEG energy both after intraperitoneal (i.p.) and intracerebroventricular (i.c.v.) application. Similar effects were observed after i.p. and i.c.v. application of the competitive antagonist, DL-CPP-ene. Whereas the MK-801 was more potent after i.p. application, DL-CPP-ene was more potent after i.c.v. administration. Intracerebroventricular administration of MK-801 and DL-CPP-ene resulted in similar changes in the pattern of local cerebral glucose utilization in the olfactory tubercle and regions of the limbic system such as the anteroventral thalamus, hippocampus and entorhinal cortex. Intravenous (i.v.) administration of MK-801 induced increases in glucose metabolism similar to those observed after i.c.v. application. In contrast, i.v. administration of DL-CPP-ene induced only small decreases of glucose utilization in several regions of the central sensory system. Thus the blockade of glutamatergic (NMDA) transmission results in decreased hippocampal EEG activity which is paralleled by increased metabolic activity in this area. We conclude from EEG recordings and [14C]2-deoxyglucose uptake experiments that both non-competitive and competitive NMDA antagonists produce the same pattern of alterations after i.c.v. administration. Apparent differences in efficacy after peripheral administration may be largely due to differences in bioavailability.


Post-traumatic selective stimulation of glycolysis.

Ventilated, chloralose anesthetized cats were subjected to 3.2 atmosphere fluid percussion injury and compared with sham operated controls. In both groups, global cerebral blood flow (CBF), cerebral metabolic rate of oxygen utilization (CMRO2), cerebral metabolic rate of glucose utilization (CMRG1), and 31P magnetic resonance spectroscopy (MRS) determinations of tissue pH (pHi) phosphocreatine (PCr), and inorganic phosphate (Pi) were measured before and 5, 15, 30, and 60 min post-trauma. Following trauma, pH decreased slightly 30 min following trauma and gradually returned to normal. CMRO2, CBF, and arterial pO2 remained at control values throughout the experiment while CMRG1 increased 243% at 5 min post-injury then rapidly returned to control values. This combination of acidosis and increased glucose utilization indicates that trauma causes a transient isolated increase in cerebral glycolysis and may be important to consider when investigating the etiology of post traumatic acidosis.


Development of neural lobe-like neurovascular contact regions after intrahypothalamic transection of the hypothalamo-neurohypophysial tract.

Fifteen days after bilateral transection of the hypothalamo-neurohypophysial tract at the level of the lateral retrochiasmatic area, neurovascular contact regions had developed proximal to 66% of the lesions. Contact regions developed in every case when neural lobe explants were placed into the lesions, and near approximately half of the lesions into which small pieces of sciatic or optic nerve were transplanted. Neurovascular contact regions were characterized by microvascular networks surrounded by dense neurophysin-immunoreactive plexuses. At the fine structural level, the organization of such regions resembled that of the neural lobe, with the single exception that capillaries were not fenestrated. Numerous neurosecretory axons were present, and palisades of neurosecretory axon terminals abutted perivascular basal laminae. Lamellopodia from glial cells partially ensheathed regenerating neurosecretory axons and often lay between terminals and the perivascular basal lamina. Terminals with many microvesicles and few neurosecretory granulated vesicles provided morphological evidence of hormone release.


Functional compartmentalization of energy production in neural tissue.

Previous work in our laboratory has shown that neural trauma results in a disparity between oxidative and glycolytic rates. In non-neural tissue, glycolysis and oxidative phosphorylation have been shown to work independently of one another, a phenomenon known as "energy compartmentalization". We believe that functional compartmentalization of energy production may also occur in the brain with glycolysis providing energy for membrane bound ionic pumps. Spreading depression, induced in rodent brain by topical KCl application, results in K+ shifts. The restoration of K+ gradients is accomplished by energy dependent Na(+)-K+ pumps. If these pumps depend upon glycolysis, blocking glycolysis should prevent reconstitution of normal [K+]e levels. The present series of experiments were designed to suggest that energy compartmentalization may also exist in brain, and that glycolytic energy production is preferentially used by Na(+)-K+ pumps to maintain normal ionic homeostasis by observing the dynamics of spreading depression induced K+ shifts before and after glycolytic blockade. Spreading depression was associated with increased K+ (48.6 +/- 16.6 mM over control) that normalized within 2.9 +/- 0.3 minutes. Following superfusion with a glycolytic blocking agent, spreading depression produced similar increases in [K+]e (40.6 +/- 12.0 mM over control) but time for reconstitution of the normal [K+]e was 400% longer than controls (2.9 +/- 0.3 to 14.9 +/- 2.1 minutes, P less than 0.001). Time required for recovery of EEG was identical pre- and post-blockade. We believe these data suggest that energy compartmentalization may exist in neural tissue and that glycolytic pathways of energy production are functionally tied to membrane Na(+)-K+ pumps.(ABSTRACT TRUNCATED AT 250 WORDS)


Selective distribution of the 66-kDa neuronal intermediate filament protein in the sensory and autonomic nervous system of the guinea-pig.

The immunohistochemical distribution of a recently identified 66-kDa neurofilament protein (NF-66) was investigated in peripheral and autonomic ganglia of the guinea-pig where it has been previously established that other neuronal intermediate filament proteins have a selective distribution. NF-66 immunoreactivity was observed in distinct subpopulations of neurons and did not coexist completely with either the neurofilament triplet or a 57-kDa intermediate filament protein (peripherin). NF-66 labelling was identical to that observed with an antibody to a 150-kDa intermediate filament or associated protein (CH1). These results further demonstrate that different neuronal intermediate filament proteins are present in selective subpopulations of neurons and that these proteins are, therefore, likely to have cell type-specific roles.


Serotonin preferentially hyperpolarizes capsaicin-sensitive C type sensory neurons by activating 5-HT1A receptors.

The effects of serotonin (5-HT) were investigated by intracellular recording from 179 dorsal root ganglion (DRG) cells classified by conduction velocity. Bath applied 5-HT depolarized 82% and hyperpolarized 4% of the A-type cells. In C-type cells, 5-HT depolarized only 41%, but hyperpolarized 39% of the cells. The depolarizing responses were of two types; an increase or decrease in R(in), mediated by 5-HT2or3 receptors, respectively. These receptors were observed in both A- and C-type cells. Hyperpolarizing responses were largely confined to A(delta)- and C-type cells. Carboxamidotryptamine and 8-OH-dipropylamino-tetralin were full agonists in eliciting hyperpolarization, and metitepin, spiperone and spiroxitrine behaved as competitive antagonists. This indicated that hyperpolarization was mediated by a 5-HT1A receptor. A 5-HT1A&3 receptor were found co-localized on some C-type cells. A strong depolarizing response to capsaicin was observed in the subgroup of C-type neurons that were also hyperpolarized by 5-HT. Thus a co-localization of capsaicin and 5-HT1A receptors was also observed.


Functional correlates of selective long-term potentiation in the olfactory cortex and olfactory bulb.

High-frequency stimulation of the granule cell layer of the olfactory bulb (OB) has previously been shown to result in a selective long-term potentiation (LTP) of late components of potentials evoked in the OB and piriform cortex (PC). The functional impact of this potentiation was explored in male Long-Evans rats with chronically implanted electrodes by comparing the effects of paired-pulse stimulation of the OB in potentiated and control animals. Effects were examined on two components of the potential evoked in the PC: A1, which represents the population EPSP produced by OB mitral cells in PC pyramidal cells via the lateral olfactory tract (LOT), and B1, which represents the subsequent population EPSP produced by PC pyramidal cells in other pyramidal cells. Two separate functional correlates of selective LTP were found. First, there was enhanced paired-pulse depression of B1, indicating increased inhibition of PC pyramidal cells. Second, there was a shift from paired-pulse facilitation to depression of A1, which was accompanied by a decrease in amplitude of the LOT volley, indicating that fewer mitral cells were activated by the stimulation. This shift was most prominent in animals with stimulating electrodes closest to the mitral cell layer, suggesting that it is dependent upon direct stimulation of mitral cell somata. These observations, together with other results reported in the manuscript, support the conclusion that there is an enhanced inhibition of mitral cells following selective LTP. Thus a primary consequence of selective LTP appears to be enhanced inhibition of principal neurons in both the PC and OB. These findings are consistent with our previous proposal that selective LTP represents potentiation at excitatory synapses made by PC pyramidal cells on inhibitory interneurons in the PC and OB.


Transforming growth factor-beta 1 differentially regulates proliferation and MHC class-II antigen expression in forebrain and brainstem astrocyte primary cultures.

To facilitate investigation of cytokine regulation of reactive astrogliosis, primary astrocyte cultures from neonatal murine forebrain and brainstem were established. Forebrain and brainstem astrocytes proliferated at a similar rate under basal culture conditions, and both were growth-inhibited by treatment with recombinant murine interferon-gamma. The growth of cultured brainstem astrocytes was significantly enhanced by exposure to recombinant human transforming growth factor-beta 1. In contrast, proliferation of forebrain astrocytes was not significantly affected by transforming growth factor-beta 1. The disparate responses of brainstem and forebrain astrocytes to transforming growth factor-beta 1 treatment were not limited to effects on cell growth, since transforming growth factor-beta 1 could block interferon-gamma-induced MHC class-II antigen expression on cultured brainstem astrocytes but not on forebrain cells. Results could not be attributed to use of an heterologous cytokine/cellular target system, since similar variability in transforming growth factor-beta 1 modulation of major histocompatibility complex antigen expression could be demonstrated using two human astrocytoma cell lines. This report is the first to document mitogenic response to transforming growth factor-beta 1 for neuroepithelial cells. The role of transforming growth factor-beta 1 in regulating aspects of reactive astrogliosis, particularly in the context of inflammatory demyelination, requires further investigation. Furthermore, these studies may provide insight into regional variability in the sequelae of inflammation within the central nervous system.


Two types of neuroplasticities in the kindling phenomenon: effects of chronic MK-801 and methamphetamine.

Using the low-frequency kindling technique, we studied the effects of chronic MK-801 and chronic methamphetamine (MAP) administration on hippocampal kindling seizure development. In experiment 1, MK-801 (0.05, 0.1 mg/kg i.p.) was administered 2 h before each electrical stimulation until kindling developed into stage-3 seizure. In experiment 2, we started daily electrical stimulations two weeks after the last injection of chronic MAP administration (6 mg/kg/day, 14 days). The number of stimulating pulses required for the triggering of epileptic afterdischarge (pulse-number threshold, PNT) was used as an indicator of the seizure threshold. PNT, afterdischarge duration (ADD) and behavioral seizure stage (BSS) of each induced seizure in the initial stage of kindling; the kindling rates for stage 3 and stage 5 seizures; seizure parameters at the completion of kindling of the drug-treated groups were recorded and compared to the values of each saline-treated control group. Our result showed that MK-801 administration prior to each electrical stimulation selectively and significantly increased PNT in the initial stage of kindling without affecting other seizure parameters. Chronic pretreatment of MAP caused a selective and significant decrease of PNT of the first two stimulations in the kindling process. Taken together with the previous studies, these results suggest that long-term potentiation plays an important role in the development of the excitability of seizure focus but not of the induced seizure's propagation in the hippocampal kindling phenomenon. Clinically MK-801 seems to be a more efficacious drug in preventing the induction of seizures than in suppressing the acquired seizures.


Effects of septal and/or raphe cell suspension grafts on hippocampal choline acetyltransferase activity, high affinity synaptosomal uptake of choline and serotonin, and behavior in rats with extensive septohippocampal lesions.

At 31 days of age, Long-Evans female rats sustained aspirative lesions of the septohippocampal pathways and, 14 days later, received intrahippocampal suspension grafts prepared from the region including the medial septum and the diagonal band of Broca (Group S, n = 11), from the region including the mesencephalic raphe (Group R, n = 11) or from both regions together (Group S+R, n = 11). Sham-operated (Group Sham, n = 9) and lesion-only (Group Les, n = 11) rats served as non-grafted controls. Seven Sham, 7 Les and 8 rats from each transplant group were tested for home cage activity (6 months after grafting) and radial maze performance (between 7.5 and 8.5 months post-grafting). One month after completion of behavioral testing, the dorsal hippocampi of these rats were prepared for measuring choline acetyltransferase (ChAT) activity and high affinity synaptosomal uptake of both [3H]choline and [3H]serotonin. The remaining rats were used for histological verifications on brain sections stained for acetylcholinesterase (AChE). The lesions increased locomotor activity, impaired radial maze learning and, in the dorsal hippocampus, reduced AChE positive staining, decreased ChAT activity (-73%) as well as high affinity uptake of both choline (-81%) and serotonin (-82%). Neither type of transplant produced any significant behavioral recovery. However, septal transplants increased hippocampal AChE positivity, restored ChAT activity and enhanced choline uptake to 116% and 70% of the values found in sham-operated rats, respectively; they had no significant effect on uptake of serotonin. Transplants from the raphe region had weak effects on hippocampal AChE positivity, increased both the ChAT activity and the choline uptake to 70% ad 38% of the sham-operated rats, respectively, and produced an (over)compensation of the serotonin uptake which reached 324% of the values found in sham-operated rats. The co-transplantation of both regions resulted in restoration of ChAT activity (113% of sham-operated rats values), choline uptake (83% of sham-operated rats) and serotonin uptake (129% of sham-operated rats). Our neurochemical data show that after extensive denervation of the hippocampus, intrahippocampal grafts of fetal neurons may foster a neurotransmitter-specific recovery which depends upon the anatomical origin of the grafted cells: a graft rich in serotonergic neurons overcompensates the serotonergic deficit, a graft rich in cholinergic neurons attenuates the cholinergic deficit, whereas a mixture of both types of grafts produces recovery from both types of deficits. Thereby, both the feasibility and the interest of the co-grafting technique are confirmed.(ABSTRACT TRUNCATED AT 400 WORDS)


Peripheral control of the antagonist muscle during unexpectedly loaded arm movements.

Ten subjects completed a series of goal-directed arm flexion movements unexpectedly perturbed by three different types of mechanical load. Examination of electromyograph (EMG) waveforms and kinematic information collected during randomly distributed test trials facilitated investigation into the interaction between loading conditions and the response-associated EMG innervation patterns. Results of the EMG waveform analysis revealed that inertial and spring loads produced cocontraction and triphasic activation patterns, respectively. Unexpected application of a stretched-spring load, which produced a change in initial torque values without changing the rate of loading, also resulted in the use of a triphasic activation pattern. These different EMG patterns were observed while movement displacement for all three loads fell within the limits of the target area.


Recovery of consummatory feeding behavior after bilateral lesions of the cerebral-buccal connectives in Aplysia californica.

In the sea hare, Aplysia californica, consummatory feeding behavior is selectively abolished by bilateral crushes of the cerebral-buccal connectives and recovers by postlesion day 13. Recovered biting responses are initially weak and increase in magnitude gradually with time. The lesions do not affect appetitive feeding behavior or unrelated reflexive behaviors. Thus, feeding in Aplysia can be used to examine the neural basis of behavioral recovery after CNS injury.


The action of CGS-19755 on the redox enhancement of NMDA toxicity in rat cortical neurons in vitro.

The effects of the competitive N-methyl-D-aspartate receptor antagonist CGS-19755 (cis-4-phosphonomethyl-2-piperidine carboxylic acid) were studied in cultures of rat cerebral cortex under normal and altered redox conditions. CGS-19755 was effective in preventing delayed neuronal death produced by an acute exposure to either glutamate (500 microM) or NMDA (200 microM), but was ineffective in protecting neurons against the toxicity induced by a prolonged exposure to kainate (500 microM). We observed that the reducing agent dithiothreitol (DTT, 500 microM), could dramatically enhance toxicity and electrophysiological responses produced by 50 microM NMDA. CGS-19755 (100 microM) could effectively block both of these effects of DTT. Any toxicity produced by DTT alone was also antagonized by CGS-19755. In contrast, oxidized DTT did not enhance NMDA toxicity nor was it toxic when added alone. These results indicate that CGS-19755 is an effective and specific neuroprotectant acting at the NMDA receptor in vitro, and that the enhancement in NMDA toxicity induced by DTT is mediated by an increase in activity at this receptor complex.


Synaptic inhibition of accessory motoneurons evoked by stimulation of the trigeminal nerve in the cat.

Stimulation of the trigeminal nerve produced polysynaptic inhibitory postsynaptic potentials (IPSPs) in accessory motoneurons of the cat. This contrasts with the observation that dorsal cervical motoneurons responded with EPSPs to trigeminal stimulus. Stimulation of the rostral part of spinal trigeminal nucleus elicited di- or polysynaptic IPSPs in accessory motoneurons. Transection of the anterior funiculus at the upper cervical cord selectively abolished the IPSPs. The IPSPs were antagonized by systematically administrated strychnine but not bicuculline.


Regulation of glucocorticoid receptor immunoreactivity in the rat hippocampus by androgenic-anabolic steroids.

To determine whether androgenic-anabolic steroids (AAS) regulate glucocorticoid receptors (GR), the effects of a mixture of three commonly abused AAS (testosterone cypionate, nandrolone decanoate and boldenone undecylenate) on GR immunoreactivity (ir), were examined in hippocampi of adrenalectomized male rats. Treatment with AAS for 1 week increased nuclear GRir in pyramidal cells of CA1, and granule cells of the dentate gyrus. These findings suggest a role for GR in the mediation of some of the CNS effects of androgenic-anabolic steroids.


Effects of central administration of opioids on facial scratching in monkeys.

Epidural and intrathecal administration of opioids to humans can produce facial pruritus and scratching that is naloxone reversible. It has been proposed that opioids may act at the level of the medulla to produce facial pruritus and associated scratching behavior. We investigated the effects of mu, delta and kappa opioid-receptor agonists microinjected unilaterally into the medullary dorsal horn (MDH) on facial scratching in cynomolgus monkeys. The selective mu opioid-receptor agonist, DAMGO (3.1-25.0 ng) produced large dose-dependent, naloxone-reversible increases in facial scratches. The selective delta opioid-receptor agonist, DPDPE (1.0-5.0 micrograms) and the selective kappa opioid-receptor agonist, U-50,488H (0.1-5.0 micrograms) did not produce significant increases in facial scratching behavior. We conclude that the MDH is a site where DAMGO, a mu opioid-receptor agonist, can act to produce facial scratching in monkeys, and that the MDH is likely the site where centrally administered opioids act to produce facial pruritus in humans.


Optic nerve regeneration with return of vision through an autologous peripheral nerve graft.

The optic fiber termination layer in the contralateral optic tectum was reinnervated and useful vision was recovered in the adult frog, after successful optic nerve regeneration through an autologous peripheral nerve-bridge used to replace the optic nerve and optic chiasma. During their course through the nerve-bridge, the optic fibers were associated with Schwann cells in the usual relationship observed in peripheral nerve.


Effects of combined methysergide and mecamylamine/scopolamine treatment on spatial navigation.

In the present study, we investigated the effects of a 5-HT2 receptor antagonist, methysergide (2.5, 7.5 and 20 mg/kg), on spatial learning in saline, mecamylamine (10 mg/kg) and scopolamine (0.8 mg/kg) treated rats. Methysergide had no effect on water-maze (WM) spatial learning in rats subjected to saline or mecamylamine pretreatments. However, scopolamine-induced WM learning deficit was augmented by methysergide at doses of 7.5 and 20 mg/kg. These results further suggest (A) that cholinergic and serotonergic systems may interact in the regulation of spatial learning, and (B) that the cholinergic component of this interaction with serotonin2 receptors is mediated by muscarinic receptors, but not by nicotinic receptors.


Enduring suppression of hippocampal long-term potentiation following traumatic brain injury in rat.

This study investigated changes in synaptic responses (population spike and population EPSP) of CA1 pyramidal cells of the rat hippocampus to stimulation of the Schaffer collateral/commissural pathways 2-3 h after traumatic brain injury (TBI). TBI was induced by a fluid percussion pulse delivered to the parietal epidural space resulting in loss of righting responses for 4.90-8.98 min. Prior to tetanic stimulation, changes observed after the injury included: (1) decreases in population spikes threshold but not EPSP thresholds; (2) decreases in maximal amplitude of population spikes as well as EPSPs. TBI also suppressed long-term potentiation (LTP), as evidenced by reductions in post-tetanic increases in population spikes as well as EPSPs. Since LTP may reflect processes involved in memory formation, the observed suppression of LTP may be an electrophysiological correlate of enduring memory deficits previously demonstrated in the same injury model.


Demonstration of artifactual coupling between spinal neurons and glial cells during intracellular recording with micropipette electrodes.

Intracellular recordings with electrophysiological properties that are characteristic of both neurons and glial cells were obtained in the isolated spinal dorsal horn. Intracellular staining supports an idea that these elements may become artifactually coupled via the recording electrode. This coupling can be a gradual process, occurring over periods up to 30 min, and may have implications for the interpretation of slow, afferent-evoked potentials in some dorsal horn neurons.


High affinity [3H]ryanodine binding sites in postmortem human brain: regional distribution and effects of calcium, magnesium and caffeine.

The pharmacological properties, regional distribution and autoradiographic localization of [3H]ryanodine binding sites were examined in postmortem human brain. Analyses of binding data from labeled ryanodine titration experiments conducted in frontal cortex revealed a single class of high affinity binding sites with a Kd value of 3.6 nM and a Bmax value of 99 fmol/mg protein. In unlabeled ryanodine titration experiments, Kd and Bmax values were 6.5 nM and 132 fmol/mg protein, respectively. Binding was found to be dependent on free Ca2+ (ED50 value, 89 microM) and was decreased by 35% in the presence of 5 mM Mg2+. This Mg2+ inhibition was abolished by the addition of 10 mM caffeine. Analysis of the regional distribution of [3H]ryanodine binding in membrane preparations revealed high levels of sites in putamen and caudate nucleus, intermediate levels in hippocampus and cortex, and low levels in cerebellum. Autoradiographically, the hippocampus displayed a high density of binding sites in the CA3 region and the dentate gyrus. Ryanodine binding sites in human brain exhibit similar, but not identical binding and pharmacological characteristics to ryanodine receptors previously identified in muscle and more recently in rat and rabbit brain and accordingly may be involved in the regulation of intracellular calcium.


Basic fibroblast growth factor (bFGF) immunolocalization in the rodent outer retina demonstrated with an anti-rodent bFGF antibody.

The distribution of bFGF in mouse and rat retinas was established using an antibody generated against a rat specific bFGF sequence. The patterns established with this antibody were distinctly different in comparison to the immunostaining patterns using antibodies prepared against similar regions of bovine and human bFGF sequences. Using the anti-rat antibody, at postpartum day 5, both species demonstrate bFGF-like labeling in the outer retina, primarily associated with developing photoreceptors. The anti-rat bFGF antibody in the adult mouse produces intense labeling at the level of the photoreceptor outer segment, whereas in the adult rat, Müller's cells and RPE were intensely labeled but the photoreceptors appeared to be unlabeled. In contrast, the bovine and human antibodies consistently label ganglion cells in both mouse and rat retinas, cells in the inner aspect of the inner nuclear layer and developing horizontal cells, whereas no photoreceptor labeling was observed. These results suggest that minor species differences in a short segment of the bFGF molecule used to generate these antibodies may result in major differences in apparent bFGF-like immunolocalization.


Puberty acceleration in female mice induced with a partially purified male urine extract: effects on catecholamine release from the olfactory bulbs and hypothalamus.

In the present experiment peri-pubertal female mice were treated with a partially purified puberty accelerating urine extract (PAUE). Mice treated with the PAUE showed an advance in the onset of puberty as indicated by significantly increased uterine weights. Treatment with the PAUE did not alter basal or potassium- (K+, 30 mM) stimulated release of catecholamines (dopamine or norepinephrine) from either anterior or posterior superfused olfactory bulb tissue fragments. There was, however, an overall significantly greater amount of basal and K(+)-stimulated release of NE from the posterior vs. the anterior olfactory bulb. Potassium-stimulated-, but not basal, release of catecholamines from the medial basal hypothalamus of PAUE-treated female mice were increased, with dopamine showing a statistically significant difference compared to water-treated females. These data demonstrate that treatment with the PAUE is a very effective means to accelerate the onset of puberty and results in accompanying increases in catecholaminergic activity, in particular dopamine, within the medial basal hypothalamus.


Characterization and distribution of angiotensin II binding sites in fetal and neonatal astrocytes from different rat brain regions.

Although angiotensin II (Ang II) binding sites have been extensively investigated in brain, revealing the presence of both AT1 and AT2 subtypes in various areas, the question as to which cells express AT1 and AT2 sites is still open. We report here that primary cultures of astrocytes obtained from various brain regions of fetal (F17) and one-day-old rats express Ang II binding sites belonging only to the AT1 subtype. The binding sites have the same binding profile in all regions tested; however, much less binding was observed in membranes of astrocytes derived from cortical than from subcortical regions and almost none were found in neonatal cortex. In addition, the dispersion method used at the onset of culture affects the number of binding sites present at the end of the culture period.


Structural asymmetry in the thalamofugal visual projections in 2-day-old chick is correlated with a hemispheric difference in synaptic density in the hyperstriatum accessorium.

Differences in visual discrimination ability between the left and right eyes of chicks, which are most prominent in young males, may result from a structural asymmetry in the organization of the visual projections from the thalamus to the visual Wulst. This asymmetry in projections is no longer present by 21 days in males when the contralateral projections from the right thalamus to the left hyperstriatum have developed. Since the asymmetry of the thalamo-hyperstriatal system results in a differential input of fibres to regions of the hyperstriatum which in turn project to the hyperstriatum accessorium (HA), one of the major differences expected within this region would be an asymmetry in the numerical density of synapses (Nv.syn/microns3). When this was examined in the hyperstriatum accessorium of 2-day-old male chicks, the density of synapses in the right HA was found to be significantly higher (22%, P less than 0.05) than in the left HA. The consequences of this asymmetry in synaptic density in the HA could be widespread and influential within the chick visual system.


Lewy bodies contain beta-amyloid precursor proteins of Alzheimer's disease.

To assess the contribution of Alzheimer's disease amyloid proteins to cortical and substantia nigra Lewy bodies (LBs), regions of postmortem brain rich in intraneuronal LBs were examined immunohistochemically. Antibodies to epitopes in domains outside the amyloidogenic beta-amyloid peptide (BAP) in BAP precursor proteins (BAPPs) as well as to the BAP itself were used as probes. These studies showed that only BAPP epitopes outside the BAP were present in substantia nigral and cortical LBs. Thus, non-amyloidogenic domains of BAPPs may be associated with intraneuronal inclusions comprised of neurofilament proteins.


Postnatal exposure to tones alters the tuning characteristics of inferior collicular neurons in the rat.

Young rats were exposed to either a 4 or 20 kHz steady tone during the first 3 weeks after birth, and then single unit responses to pure tones were studied at their inferior colliculi (IC). The best frequencies (BF's) of the cells showed strong clustering around the frequency of exposure.


Divergent effects of deep cerebellar lesions on two different conditioned somatomotor responses in rabbits.

Rabbits with bilateral lesions involving either the anterior interpositus nucleus or the superior cerebellar peduncle were subjected to appetitive Pavlovian conditioning training involving repeated pairings of a 2-s tone with an intraoral pulse of water. Such training resulted in the rapid development of robust, anticipatory jaw-movement responses (JM CRs) to the tone, and, in fact, the performance levels exhibited by lesioned animals did not differ significantly from those observed in sham-operated control animals. Additional experiments involving unpaired tone/water presentations confirmed the associative character of the JM CRs. On the other hand, lesioned animals exhibited severe bilateral performance deficits when later subjected to aversive eyeblink conditioning procedures, consistent with previous findings. The present results thus suggest that the interpositus nucleus is not an essential neural substrate for the development of appetitively conditioned masticatory responses.


Reappearance of calcitonin gene-related peptide-like immunoreactivity in the dorsal horn in the long-term dorsal root transected rat.

Calcitonin gene-related peptide (CGRP)-immunoreactive (IR) fibers in the rat dorsal horn superficial laminae vanish almost completely 3 weeks following unilateral dorsal rhizotomy. After a prolonged survival (20 weeks) of dorsal rhizotomy there is, however, a reappearance of CGRP-IR fibers in the corresponding laminae of the dorsal horn. The density of such IR fibres showed a clear gradient with the lowest number found in the midlesion region and an increase in density towards the neighboring, intact segments. In normal as well as lesioned rats, no neurons intrinsic to the dorsal horn contained detectable levels of CGRP-like immunoreactivity (LI). Furthermore, no cells could, by use of in situ hybridization, be demonstrated to contain detectable levels of mRNA encoding for CGRP in the dorsal horn. Based on these findings, we suggest that the CGRP-IR fibers observed following long-term survival of dorsal rhizotomy derive from proliferating collateral branches of primary afferents of neighboring intact segments.


Intracellular signalling systems controlling the 5 alpha-reductase in glial cell cultures.

Glial cells are able to metabolize testosterone into DHT through the action of the enzyme 5 alpha-reductase. DHT may be further processed to 3 alpha-diol by the 3 alpha-hydroxysteroid-dehydrogenase. The aim of this study was to analyze if a modulation of two second messenger systems might be able to modify the 5 alpha-reductase and the 3 alpha-hydroxysteroid-dehydrogenase activities present in glial cells. To this purpose, the formation of DHT has been measured in rat glial cell cultures after different time of exposure to TPA, 4 alpha-Ph, an active and an inactive phorbol ester respectively, and 8-Br-cAMP. The results obtained indicate that the formation of DHT is not modified by the addition of phorbol esters. On the contrary, a statistically significant increase of 5 alpha-reductase activity, over control levels, has been observed after 6, 12, and 24 h of incubation with 8-Br-cAMP (10(-3) M). The effect of the cAMP analogue appears to be specific for the 5 alpha-reductase, since the 3 alpha-hydroxysteroid-dehydrogenase did not show any variation after exposure to the drug. In conclusion, the present data suggest that proteinkinase A (PKA) might be involved in the control of the 5 alpha-reductase in glial cells. It is postulated that nervous inputs utilizing cAMP as the second messenger might modify the activity of this enzyme in glial cells.


Differential actions of acetylcholinesterase on the soma and dendrites of dopaminergic substantia nigra neurons in vitro.

In the substantia nigra, acetylcholinesterase (AChE) has non-cholinergic action on dopaminergic neurons. The subset of neurons particularly sensitive to AChE are characterized by functionally active apical dendrites extending into the pars reticulata and generating a powerful calcium conductance. This study thus attempted to establish directly the importance of these dendrites regarding the action of AChE. Segregation of the pars compacta from the pars reticulata did not affect the AChE-induced hyperpolarization on this sub-set of dopaminergic neurons. However, the ionic basis of the hyperpolarization was related to the integrity of the neurons: AChE caused an opening of potassium channels in intact cells. On the other hand when the pars reticulata containing apical dendrites was removed, an action of AChE involving the closure of calcium/sodium channels was revealed. The results demonstrate that the net effect of AChE need not be related to any particular segment of the dopaminergic neurons, whereas the nature of the mechanism underlying that effect depends on the presence, or otherwise, of the apical dendrites.


Fluoxetine attenuates the DL-fenfluramine-induced increase in extracellular serotonin as measured by in vivo dialysis.

Rats with hippocampal dialysis probes were treated with DL-fenfluramine (FEN), fluoxetine, or FEN with fluoxetine pre-treatment. FEN (12.5 mg/kg) increased extracellular serotonin (5-HT) from 0.4 +/- 0.04 to 25.2 +/- 4.16 pg/10 microliters. Fluoxetine (10.0 mg/kg) increased extracellular 5-HT levels from 0.4 +/- 0.05 to 2.4 +/- 0.33 pg/10 microliters. FEN-induced increases in extracellular 5-HT were attenuated by 66% with fluoxetine pre-treatment. This result supports the view that the 5-HT releasing properties of FEN are mediated by the 5-HT uptake transporter.


Electron microscopic identification of lamina I axon terminations in the nucleus submedius of the cat thalamus.

Ascending lamina I axons were labeled with Phaseolus vulgaris leucoagglutinin and the synaptic connections of their terminals in nucleus submedius (Sm) were studied in the electron microscope. The terminals were large, contained rounded synaptic vesicles, and were involved in complex synaptic aggregations with pre- and postsynaptic dendrites. It was observed that clustered large boutons from a single axon could contact a single dendritic shaft. These observations support a sensory role for lamina I input to Sm.


Axon types classified by morphometric and multivariate analysis in the rat optic nerve.

Calibers of the rat optic nerve axons distribute unimodally and it is difficult to distinguish groups among them. However, these fibers arose from 3 types of ganglion cells and showed 3 conduction velocities. Performing a cluster analysis over several ultrastructural parameters we found 3 main groups of fibers. These groups are present in a very similar proportion to the ganglion cells groups described in the rat retina.


Enhanced responses of nucleus accumbens neurons in male rats to novel odors associated with sexually receptive females.

One group of male rats was trained to associate novel odors with three different environmental conditions: the presence of (i) a sexually receptive female (RF), (ii) an unreceptive female (UF) and (iii) no other rat (NO). A second group of males received no training. Single units in nucleus accumbens (NAC) were then recorded in anesthetized animals and their responsiveness to various odors was tested. Odors that had been associated with receptive females during training evoked significantly more unit responses in NAC than did the same odors in untrained males. There were no differences between trained and untrained males in the numbers of units responsive to odors associated with unreceptive females and with the empty training chamber. In trained animals, both the percentage of responding units and the magnitude of olfactory-evoked responses were significantly larger with RF-associated odors than with either UF or NO odors. Both of these effects were more pronounced in rats that had ejaculated with females during training than in rats that had not. Findings demonstrated that pairing odors with the presentation of sexually receptive females enhanced the responsiveness of NAC neurons to those odors and indicated a role for NAC in associating environmental stimuli with natural reward processes.


The cell recognition molecule, cognin, mediates choline acetyltransferase activity in embryonic chick retina.

Cell signaling and cell-cell interactions play an important role in neuronal differentiation in the embryonic CNS. Previous work (Hausman, R.E., Vivek Sagar, G.D. and Shah, B.H., Dev. Brain Res., 59 (1991) 31-37) had shown that cholinergic differentiation in the embryonic chick retina depends on insulin and neuron-neuron interactions. Here, we pursued the molecular nature of that dependence on cell interactions. The embryonic chick retina is known to contain several cell adhesion or recognition molecules. We asked if retina cognin, a 50 kDa cell surface-associated protein, played a role in controlling cholinergic differentiation in the developing chick retina. As previously, cholinergic differentiation was measured by two markers: choline acetyltransferase (ChAT) activity and high-affinity choline uptake. We used polyclonal antibody to cognin to determine if blocking cognin-mediated cell interactions would affect the normal embryonic increases in these cholinergic markers. We demonstrated a 40% inhibition of the normal developmental appearance of ChAT activity in retina neuronal cultures from early development, but no effect in cultures from more differentiated retina. The inhibition was selective for retina, since it was not seen in neural tissues like cerebrum and cerebellum that also express ChAT. In contrast to the effect of insulin, choline uptake was not affected by treatment with cognin antibody. Antibodies to two other cell recognition molecules present in the retina (Ng-CAM and N-cadherin) did not block the normal developmental appearance of ChAT. These results suggest that cognin-mediated interactions play a unique role in the control of one aspect of cholinergic differentiation in the developing chick retina.