Functional regeneration of glossopharyngeal nerve through micromachined sieve electrode arrays. To assess the potential of micromachined silicon sieve electrodes for long term recordings from single afferent sensory fibers, we implanted them between the cut ends of rat glossopharyngeal nerves which innervate taste and somatosensory receptors on the posterior tongue. After the implants had been in place for an average of 101 days nerve regeneration was measured using histological and electrophysiological methods. Axons of the glossopharyngeal nerve regenerated through holes in the sieves and supported the functional regeneration of taste, thermal and mechanoreceptors.


Astringent compounds suppress taste responses in gerbil. Astringent tastes are generally considered those that induce long-lasting puckering and drying sensations on the tongue and membranes of the oral cavity. Electrophysiological recordings were made here from the whole chorda tympani nerve in gerbil to understand the interactive effect of astringent-tasting molecules with a broad spectrum of tastants including mono- and divalent salts, bitter compounds, acids, and sweeteners. The astringent tasting compounds were tannic acid (24 mM at pH's 2.9 and 5.5), aluminum ammonium sulfate (30 mM), aluminum potassium sulfate (10 mM) and gallic acid (30 mM). Hydrochloric acid (1 mM, pH 2.9) was also tested to control for acidity, since aqueous solutions of astringent-tasting compounds are acidic. Adaptation of the tongue to tannic acid (24 mM) at both pH 2.9 and 5.5 markedly inhibited responses elicited by salts, acids, sweeteners, and bitter-tasting compounds. The degree of the inhibition at these two pH values is about the same which suggests that tannic acid itself (as opposed to acidity) may produce this inhibition. Chorda tympani responses to sweeteners were completely suppressed by tannic acid; responses to KCl, NH4Cl, and urea were the least suppressed. The aluminum salts also inhibited the chorda tympani responses to all stimuli tested. Gallic acid, which is weakly astringent, had minimal effects on the chorda tympani responses to the test compounds. These data suggest that both tannic acid and the aluminum salts inhibit a variety of transport pathways and receptors in taste cells for a broad spectrum of tastants. The inhibition of some of these pathways may contribute to the astringent taste sensation.


Evocation of postural atonia and respiratory depression by pontine carbachol in the decerebrate rat. To study mechanisms underlying the postural muscle atonia and respiratory depression associated with rapid eye movement (REM) sleep, the cholinergic agonist, carbachol, was microinjected into the pontine reticular formation in decerebrate, spontaneously breathing rats. Carbachol injection led to a simultaneous depression of neck and tonic intercostal EMG activity (lasting 14.5 min +/- 7.6 (S.D.)) and a reduction of the respiratory rate. Phasic inspiratory intercostal activity was not consistently depressed. After a spontaneous recovery from the atonia and respiratory depression, subsequent carbachol injections at the same site produced similar responses. Thus, the decerebrate rat may provide a useful model for studies of the inhibitory neural mechanisms activated by the increased acetylcholine levels in the pons that occur in REM sleep. Pontine carbachol effects in rats differ from those described in cats in a manner analogous to differences in the patterns of natural REM sleep in these two species.


The internal carotid artery has a sleeve of increased innervation density within the cavernous sinus in monkeys. The numerical density of nerve terminals of the internal carotid artery was measured using interrupted serial sections and compared with densities sampled from the major cerebral arteries of cynomolgus and rhesus monkeys. In its course through the carotid canal and the foramen lacerum the artery received few terminals. Nerve terminal density increased substantially within the cavernous sinus in 13 of 19 animals, reaching a peak shortly before emerging to join the circle of Willis. The density dropped rapidly on leaving the sinus. The increase was present in both species and rose to a mean nerve terminal density at least ten times that of any other artery measured. In 6 monkeys terminal incidence was unchanged through the sinus. The possible relevance of the nerve terminal sleeve to cerebral vasodynamics and to vascular head pain is discussed.


Characterization of phospholipase D in a cell-free system of cultured cells derived from rat frontal cortex. The existence and regulation of phospholipase D (PLD) activity in cell-free system from primary cultured cells of fetal rat frontal cortex were investigated. PLD activity was detectable only in the presence of Triton X-100. Other detergents examined (deoxycholate, taurocholate, CHAPS, Tween 20, sodium dodecyl sulfate) caused only a small increase in PLD activity. Triton X-100 enhanced PLD activity maximally at 0.1% (w/v) and reduced at higher concentrations. The optimal pH was about 7.2. Both Ca2+ and Mg2+ inhibited PLD activity in a dose-dependent manner. When comparing the primary cultured cells with adult rat frontal cortices, all of the results of the primary cultured cells were in agreement with those of the frontal cortices. Moreover, the apparent Km value of the enzyme in primary cultured cells for phosphatidyl-choline was the same as that in rat frontal cortex. These results suggest that the same kinds of PLD exist in the primary cultured cells and the rat frontal cortex, and that the primary cultured cells are a good experimental model for analyzing the mechanism of PLD in neuronal system.


Selective enhancement by basic fibroblast growth factor of NMDA receptor-mediated increase of intracellular Ca2+ concentration in hippocampal neurons. The short-term effect of bFGF on intracellular Ca2+ concentration ([Ca2+]i) of hippocampal neurons was investigated using dissociated cell cultures. Changes in [Ca2+]i were measured by microfluorometrically monitoring the fluorescence intensities from individual neurons loaded with fura-2. Perfusion of bFGF (20 ng/ml) alone did not affect the basal level of [Ca2+]i in hippocampal neurons, but clearly enhanced the [Ca2+]i increase induced by NMDA. Quisqualate or KCl-induced [Ca2+]i increase was not influenced by bFGF. These results suggest that bFGF selectively enhances the NMDA receptor-mediated response in hippocampal neurons.


Immunocytochemical localization of parvalbumin- and neurofilament triplet protein immunoreactivity in the cat retina: colocalization in a subpopulation of AII amacrine cells. Using antibodies against parvalbumin and neurofilament triplet protein, colocalization of these two neuronal markers was revealed in all of type A horizontal cells and alpha ganglion cells and in a small number of AII amacrine cells of the cat retina. Besides the double-labeled neurons, parvalbumin alone was present in type B horizontal cells, in small numbers of starburst- and A13-like amacrine cells and in the somata of unidentified ganglion cells. The processes of the double- or single-labeled amacrine cells did not have a continuous retinal cover. Although the parvalbumin- and neurofilament-immunolabeled amacrine cells belonged to groups of neurons with well-defined cell morphologies, their neurochemical features differed from other AII, starburst and A13 amacrine cells. The presence of these cells may be due to an accidental expression of an unusual combination of neurochemical features during retinal development. It is also possible that these cells support the functioning of ganglion cells with rarely occurring complex receptive fields.


Low sodium injury in the hippocampal slice is mediated through NMDA receptors. Perfusion of hippocampal slices with normoxic medium containing no added sodium resulted in a rapid loss of the CA1 population spike, with only 35 +/- 10% (S.E.M.) (P < 0.001) recovery after a 15 min exposure. This injury was prevented by the non-competitive N-methyl-D-aspartate (NMDA) antagonist MK-801 (93 +/- 4% recovery, P < 0.001), suggesting that low sodium injury may be mediated by opening of the NMDA receptor-associated ionic channels, possibly secondary to the well known sodium dependency of transmitter uptake systems. By contrast, slice perfusion with a medium moderately low in sodium (26 mM) produced only slight injury to the CA1 population spike under normoxic conditions (76 +/- 8% recovery) and provided no protection against hypoxic injury. Low chloride medium also provided no protection against hypoxic injury.


Cyclosporin A protects against ischemia-reperfusion injury in the brain. We investigated the protective effect of Cyclosporin A (CsA) against ischemia-reperfusion injury in the brain using a transient focal ischemia model in rats. In CsA-treated rats, ischemic brain edema formation 1 day after reperfusion in the cerebral cortex perfused by the middle cerebral artery (MCA) and infarct size were decreased compared with those in olive oil treated control rats. These results suggest that CsA is beneficial in reducing ischemia-reperfusion injury, possibly by the suppression of immunological reactions.


Convergent serotonin and GABA innervation of VIP neurons in the suprachiasmatic nucleus demonstrated by triple labeling in the rat. By means of a combination of serotonin (5-HT) uptake radioautography and dual immunocytochemistry, vasoactive intestinal polypeptide (VIP) neurons in the suprachiasmatic nucleus are demonstrated to simultaneously receive both 5-HT and GABA afferents at their somatic and dendritic levels. These data constitute a further step towards the improved characterization of the morphological substrate of the integrative function of these neurons, which are known to play an important role in the delivery of light-mediated rhythmic signals to other parts of the brain.


Nitric oxide synthesis couples cerebral blood flow and metabolism. The most fundamental aspect of the cerebral circulation is the well-described coupling of cerebral metabolic activity and cerebral blood flow. A number of substances have been proposed to link flow and metabolism, including K+, pH and adenosine. In the alpha-chloralose anaesthetised cat we studied simultaneously cerebral neuronal activity and local blood flow to attempt to dissociate the two and thus determine the coupling substance. Neuronal activity was determined by monitoring unit firing in the parietal cortex with tungsten in glass microelectrodes while local cerebral blood flow in the same area was monitored continuously using laser Doppler flowmetry. To initiate an increase in metabolic activity and, pari passu, blood flow spreading depression was elicited by needle stick injury. Spreading depression when initiated causes a wave of depolarization, measured as an increased firing rate and associated marked (400 +/- 95%) increase in local cerebral blood flow. Intravenous administration of NG-nitro-L-arginine methyl ester (1-NAME), a potent nitric oxide synthase inhibitor, produced a complete blockade of the hyperemia associated with spreading depression but no change in either resting cell firing or spreading depression-evoked increases in firing rate. These data demonstrate at least for spreading depression-elicited increases in metabolic activity, that nitric oxide (NO) is a key coupling compound that links changes in cerebral blood flow and metabolism. These data imply that NO may have a more general role in flow/metabolism coupling and further studies in other situations are required to determine the extent to which NO is responsible for this fundamental cerebrovascular phenomenon.


Circadian rhythm of cell proliferation in the telencephalic ventricular zone: effect of in utero exposure to ethanol. The neocortical ventricular zone is composed of a desynchronized population of proliferating cells. These cells give rise to neurons in the infragranular laminae of neocortex. The present study documents a diurnal rhythmicity of cell proliferation in the ventricular zone and examines the effects of ethanol on this biological clock. Pregnant rats were fed one of 3 diets. They were provided an ethanol-containing (6.7% v/v) liquid diet ad libitum between gestational day (G) 6 and G18, pair-fed an isocaloric liquid control diet, or fed chow and water. Throughout the experiments, the rats were fed either at 08.00 h (E.S.T) or at 17.00 h (lights on 06.00 to 18.00 h). Rats were given a single injection of bromodeoxyuridine (BrdU) on G17 at one point during a 24 h period (03.00, 06.00, 09.00 h, etc.). The fraction of ventricular cells that incorporated the BrdU was determined using quantitative immunohistochemical methods. Pair-fed control rats (fed at 08.00 or 17.00 h) consumed their food within 4 h of presentation. The ratio of cells passing through the S-phase of the cell cycle changed diurnally; the ratio was highest during the day (0.52 +/- 0.01 at 12.00 h) and lowest during the night (0.40 +/- 0.02 at 03.00 h). In contrast, the ethanol-fed rats grazed on their food throughout the dark cycle regardless of when the food was presented. The mean peak blood ethanol concentration was 142 +/- 13 mg/dl during the dark phase and less than 25 mg/dl during the light phase. Prenatal exposure to ethanol eliminates the fetal circadian rhythm in cell proliferation (mean labeling index of 0.45 +/- 0.03).(ABSTRACT TRUNCATED AT 250 WORDS)


Neurons in the dorsal column nuclei of the rat respond to stimulation of neck mechanoreceptors and project to the thalamus. Electrophysiological recordings were made from neurons in the dorsal column nuclei which were activated by stimulation of muscle and cutaneous receptors in the neck of the rat. 222 units were studied, 158 (71%) of which responded to activation of cutaneous mechanoreceptors while 64 (29%) were activated by muscle receptors. The response patterns of 12 neurons with input from receptors in neck muscles were tested more fully. Their response patterns strongly suggested that 6 were activated by muscle spindle afferents while the other 6 were activated by Golgi tendon organ afferents. 18 (8%) of the neck-responsive neurons in the medulla were shown to project rostrally to the thalamus.


Hypothalamic paraventricular nucleus lesions do not abolish glucoprivic or lipoprivic feeding. This experiment assessed the importance of the hypothalamic paraventricular nucleus (PVN) for feeding stimulated by blockade of glucose utilization (glucoprivic feeding) and fatty acid oxidation (lipoprivic feeding). The PVN was investigated because it is innervated by neurons residing in the area postrema and nucleus of the solitary tract (AP/NTS) region where lesions have been shown to abolish both glucoprivic and lipoprivic feeding, and because the PVN appears to be a site of action for certain feeding-stimulatory peptides and amines. Bilateral electrolytic lesions were placed in the PVN and adjacent areas. Lesioned rats were subsequently tested for feeding in response to 2-deoxy-D-glucose (2DG)-and mercaptoacetate (MA)-induced blockade of glucose and fatty acid oxidation, respectively. Results revealed that total destruction of the PVN does not impair either 2DG- or MA-induced food intake and suggest that this structure is not essential for these particular controls of feeding.


Axial muscle EMG responses evoked by cutaneous flank nerves in the female rat: effects of spinal transection, steroid hormones, and anesthesia. In the lateral longissimus muscle (LL) of ovariectomized, female rats anesthetized with low surgical doses of urethane (1.0 g/kg), cutaneous reflexes with similar EMG and response patterns could be elicited from CNS-intact rats and from rats 24 h after complete thoracic spinal cord transection. The probability of eliciting a response to contralateral cutaneous nerve stimulation alone is much lower in rats with complete spinal transections compared to CNS-intact rats. For both CNS-intact and spinal-transected rats, responses to ipsilateral cutaneous nerve stimulation had a shorter latency and required significantly less current on average than responses to contralateral stimulation. The respective currents for eliciting threshold responses to ipsi- and contralateral stimulation are less for CNS-intact than spinal-transected rats. For both CNS-intact and spinal-transected rats, responses to bilateral cutaneous nerve stimulation were inconsistent in the same animal from run to run. With the variability of response at this anesthetic level, no consistent effects of progesterone (acute, i.v.) or estrogen (acute, i.v. and pretreatment, s.c.) were observed in spinal-transected rats. Intravenous progesterone reduced early, unilateral responses in CNS-intact rats anesthetized with 1.0 g of urethane/kg. For both CNS-intact and spinal-transected rats, additional anesthesia during EMG recording produced a gradual decline in response magnitude which could be recovered with a modest increase in stimulus intensity. However, spinal-transected rats appear to require less anesthesia to reduce comparable responses. The results suggest that supraspinal input is especially effective for facilitating contralateral cutaneous reflexes in back muscles, whereas it contributes more equally with afferent input and segmental circuitry to the efficacy of ipsilateral cutaneous reflexes.


Stimulation of 5-HT1A receptors in the dorsal hippocampus impairs acquisition and performance of a spatial task in a water maze. 8-Hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT), a potent 5-HT1A receptor agonist, was infused in the dorsal hippocampus of rats and its effect on acquisition and performance of a 2-platform spatial discrimination task was studied using a water maze. The infusion (0.5 microliter/min) of 2 but not 0.4 microgram 8-OH-DPAT in the CA1 region of the dorsal hippocampus impaired rats' accuracy with no effect on latency (except day 3). At 5 micrograms 8-OH-DPAT impaired rats' accuracy and significantly increased choice latencies from day 2 to day 5 of the training period. The dose of 2 micrograms significantly increased the errors of omissions on the first day of training and animals which had received 5 micrograms 8-OH-DPAT made significantly more errors of omission on the first and second days of training. Intrahippocampal administration of 1 microgram spiroxatrine, a 5-HT1A receptor antagonist, antagonized the effect of 5 micrograms 8-OH-DPAT on accuracy and choice latency with no significant effect on the errors of omission on days 1 and 2 of training. Infusion of 2 and 5 micrograms 8-OH-DPAT in the dorsal hippocampus also impaired accuracy in well-trained rats. The results suggest that stimulation of 5-HT1A receptors in the CA1 region of the dorsal hippocampus causes an impairment of spatial discrimination in rats.


Recoverin in pineal organs and retinae of various vertebrate species including man. Recoverin is a recently discovered 26 kDa calcium-binding protein, which activates guanylate cyclase in retinal photoreceptors when the intracellular concentration of free calcium drops upon photoexcitation. In this study we examined the distribution of recoverin in retinae and pineal organs of Xenopus laevis larvae, 1-day-old chicken, adult pigeon, albino rat, sheep and man by means of immunocytochemistry. Recoverin immunoreaction was found in all species investigated except for the chicken. In the retina, recoverin immunoreaction was restricted to photoreceptors; all other cell types were immunonegative. In the pineal organ, the recoverin immunoreaction labeled 'pinealocytes of the sensory line', i.e. classical pineal photoreceptors of Xenopus laevis larvae, modified pineal photoreceptors of pigeon, and pinealocytes of mammals. The number of recoverin immunoreactive pinealocytes varied considerably among species of mammals: very few cells were stained in the rat pineal organ, whereas in rabbit, sheep and man, numerous pinealocytes were found to be recoverin-immunoreactive. No immunocytochemical staining was observed after preabsorption of the recoverin antibody with the recombinant protein. Immunoblotting experiments showed that the immunoreaction is due to a protein of 26 kDa in both retina and pineal tissue. Thus, recoverin appears to belong to the family of proteins which are expressed in both retina and pineal organ and are highly conserved in the course of phylogeny. Recoverin may be involved in phototransduction in the directly light-sensitive pineal organs of poikilothermic vertebrates and birds. However, the functional role of recoverin in the mammalian pineal organ, which is not photosensitive, remains unknown.


Selective form of an excitable membrane plasticity. This work describes the change in an active electrogenesis of the command neurons responsible for defensive closure of a snail's pneumostome during elaborating, extinction and restoration of a classical conditioned defensive reflex to a tactile stimulus. Tactile stimulations applied to different parts of a snail's body served as a differential stimulus. As the biological value of a conditioned stimulus increases due to learning, the excitability of command neurons in response to conditioned stimulus rises. At the same time the neurons demonstrated a reduced excitability in response to a differentiating stimulus.


Tactile and taste sensory maturation of the frog larvae tongue. The maturation process of organs in the oral cavity of the larvae of bullfrogs, Rana catesbeiana, corresponding to the larval stages XVIII to XXV distinguished by Taylor and Kallros Anat. Rec., 94 (1946) 7-25, were investigated. In this study, these larval stages were divided by the width of the mouth slit into A1, A2, B and C stages. The A1 and A2 stages corresponded to larval stages XVIII-XIX and XX, respectively. The B and C stages corresponded to stages XXI and XXII-XXV, respectively. In stage A1, mechanical and chemical stimulation of the tongue rudiment, in most larvae, failed to elicit afferent discharges in glossopharyngeal nerves. However, in a small number of larvae, obvious afferent discharges of these nerves were generated. In stage A2, glossopharyngeal nerves in most larvae began responding to the same stimulations. The amplitude increased with decreasing duration of afferent discharges, resembling those of the adult. In stage B, the first glossopharyngeal-hypoglossal nerve reflex (G-H reflex) was elicited by mechanical stimulation of the tongue. In stage C, the G-H reflex was also induced by chemical stimulation and the latency of the G-H reflex elicited by mechanical stimulation decreased. In this stage, expansion and extension of the tongue increased. Therefore, the natural function of this reflex for the intake of food may follow this stage.


In vivo mapping of drug-induced seizures with voltage-sensitive dye. The voltage-sensitive dye diO-C2-5 was used to produce an in vivo map of the membrane potential in two types of seizures. Mild limbic seizures were induced in rats with kainic acid; clonic convulsive seizures were induced with bicuculline. Kainic acid animals showed various levels of neural depolarization during their seizures in limbic, thalamic, cortical, and brainstem sites. The bicuculline animals showed uniformly greater levels of neural depolarization during their seizures. The magnitude of these changes relative to controls varied across seizure models and reflected the different underlying neural mechanisms for each model. The ability of the technique to capture local electrical events provides a new tool in which to explore brain activity.


Angiotensin II binding sites in the hamster brain: localization and subtype distribution. This study was designed to characterize the distribution of angiotensin II (AII) binding sites in the hamster brain. Brain sections were incubated with [125I][sar1,ile8]-angiotensin II in the absence and presence of angiotensin II receptor subtype selective compounds, losartan (AT1 subtype) and PD123177 (AT2 subtype). Binding was quantified by densitometric analysis of autoradiograms and localized by comparison with adjacent thionein stained sections. The distribution of AII binding sites was similar to that found in the rat, with some exceptions. [125I][sar1,ile8]-angiotensin II binding was not evident in the subthalamic nucleus and thalamic regions, inferior olive, suprachiasmatic nucleus, and piriform cortex of the hamster, regions of prominent binding in the rat brain. However, intense binding was observed in the interpeduncular nucleus and the medial habenula of the hamster, nuclei void of binding in the rat brain. Competition with receptor subtype selective compounds revealed a similar AII receptor subtype profile in brain regions where binding is evident in both species. One notable exception is the medial geniculate nucleus, predominately AT1 binding sites in the hamster but AT2 in the rat. Generally, the AII binding site distribution in the hamster brain parallels that of the other species studied, particularly in brain regions associated with cardiovascular and dipsogenic functions. Functional correlates for AII binding sites have not been elucidated in the majority of brain regions and species mismatches might provide clues in this regard.


Calretinin-immunoreactive neurons in the primary visual cortex of dolphin and human brains. A new class of gamma-aminobutyric acid (GABA)ergic neurons immunoreactive to the calcium-binding protein calretinin (CR) was demonstrated in primary visual cortices of the bottlenose dolphin (Tursiops truncatus) and humans (Homo sapiens). Comparative analysis revealed several differences between dolphin and human visual cortex in the laminar distribution of CR-positive perikarya, although general typology of the immunoreactive CR-positive neurons was similar in both species. Thus, in both human and dolphin primary visual cortex almost all CR-positive neurons are non-pyramidal, either fusiform or bipolar cells, oriented with their long axis along the radial axis of the cortex. Large multipolar stellate cells were also observed in layers I and VI. The CR-positive neurons in the dolphin visual cortex are concentrated almost exclusively in layer I and, to a lesser extent, in layer II. In all other layers (IIIa, b, IIIc/V and VI) of the dolphin visual cortex CR-positive neurons were only rarely seen. In the human primary visual cortex CR-positive neurons are located mainly in layers II, III and IVa, b, c, with considerably lower densities of these cells observed in layers V and VI. CR-positive neurons in layer I of the human visual cortex are represented by Cajal-Retzius horizontal cells, whereas no such cells were seen in layer I of the dolphin neocortex. The numerical density of CR-positive neurons in the dolphin primary visual cortex is significantly lower than in the area of cortex in humans.(ABSTRACT TRUNCATED AT 250 WORDS)


Neural distribution of estrogen receptor immunoreactive cells in the female musk shrew. In this report we describe the neural distribution of estrogen receptor immunoreactivity (ER-IR) in the female musk shrew (Suncus murinus). The highest concentrations of neurons containing ER-IR were found in the preoptic areas, the lateral septum, the anterior arcuate and ventromedial nuclei of the hypothalamus, the medial nuclei of amygdala, and the midbrain central grey. Additional preoptic, hypothalamic, limbic and midbrain nuclei also contained ER-IR cells. The distribution of ER-IR was similar to that described in other mammals and birds, with several important differences. In the female musk shrew, there was little difference in ER-IR intensity or distribution when brains from gonadally intact and ovariectomized musk shrews were compared. In addition, long-term treatment with a supraphysiological dose of estradiol was required to detect a decrease in ER-IR intensity. Finally ER-IR was noted in both nuclear and cytoplasmic regions of cells in ovariectomized and gonadally intact musk shrews. The dense ER-IR noted in intact females as well as the presence of cytoplasmic stain may be due to the unusual relationship between estradiol, ovulation and sexual receptivity in this species.


Prenatal stress selectively alters the reactivity of the hypothalamic-pituitary adrenal system in the female rat. A study was made of the effects of prenatal stress on the reactivity of the hypothalamic-pituitary adrenal (HPA) axis in male and female offspring. Rat dams were subjected to noise and light stress on an unpredictable basis throughout pregnancy. At 28 days of age mRNA for POMC, proenkephalin and prodynorphin were measured in the hypothalamus of the offspring. A marked reduction was found in POMC mRNA in PS females (PSF) but not in males (PSM), but the other mRNA's did not differ from controls (C). At 60 days of age, PSF has 3 times higher resting levels of serum corticosterone (COR) and significantly lower dexamethasone (DEX)3H hippocampal binding sites than CF. Overnight adrenalectomy abolished the difference in DEX binding. After 10 min exposure to open field PS males and females voided more fecal pellets and made fewer center entries than C offspring, testifying to increased emotionality. Open field stress caused a 3-5-fold rise in circulating COR in all groups within 15 min, which returned to baseline by 90 min in all rats except PSF. These data show that prenatal stress can cause permanent alterations in the behavior of both sexes in stressful situations but appears to cause a selective effect on the HPA axis in the female rat.


Hippocampectomy disrupts the topography of the rat eyeblink response during acquisition and extinction of classical conditioning. The effect of hippocampal lesions on acquisition and extinction of eyeblink conditioning in rats was examined with the procedure described by Schmajuk and Christiansen (Physiol. Behav., 48 (1990) 755-758). Male Long-Evans rats received either sham, cortical control or hippocampal lesions. Animals were trained in a delay conditioning paradigm with a 500-ms, 98-dB, white noise conditioned stimulus; a 150-ms, 5-psi air puff unconditioned stimulus; a 350-ms interstimulus interval; and a 60-s average intertrial interval. Each animal received 50 trials daily until it reached a criterion of 80% conditioned responses (CR) on a given day. After acquisition criterion was reached, extinction began the next day and continued until each animal achieved a criterion of one CR in a block of 10 consecutive trials. Although hippocampal lesions affected neither acquisition nor extinction rates, hippocampal lesioned animals showed significantly shorter CR onset latency during acquisition and extinction, and larger CR peak amplitude during acquisition. The results are discussed in terms of hippocampal modulation of cerebellar learning.


Distribution of morphine in brain regions, spinal cord and serum following intravenous injection to morphine tolerant rats. In order to determine the possible contribution of altered distribution of morphine in the morphine tolerance process, the distribution of morphine was studied in brain regions and spinal cord, following its intravenous administration. Male Sprague-Dawley rats were made tolerant to morphine by implanting 6 morphine pellets, each containing 75 mg of morphine base, for 7 days. Seventy-two hours after the removal of the pellets, a time when serum morphine levels were negligible or absent and yet tolerance to the pharmacological effects of morphine was present, morphine (10 mg/kg, i.v.) was injected in placebo and morphine pellet implanted rats. At various times (5, 30, 60, 120 and 360 min) after the injection of morphine, brain regions (hypothalamus, cortex, hippocampus, midbrain, pons and medulla, striatum and amygdala), spinal cord and serum were collected. The level of morphine in the tissues was determined by using a highly sensitive and specific radioimmunoassay (RIA) method. Five minutes after morphine injection, the concentration of morphine was the highest in the hypothalamus and the lowest in amygdala. The concentration of morphine in hypothalamus, pons and medulla, hippocampus and midbrain of morphine tolerant rats was smaller than in placebo pellet implanted rats. The tissue to serum ratio of morphine in the hypothalamus, hippocampus, striatum, midbrain and cortex were also smaller in morphine tolerant than in non-tolerant rats. The concentration of morphine in brain regions with time did not exhibit linearity. At other time intervals like 30 and 60 min, the concentration of morphine in several brain regions and spinal cord was significantly higher in morphine tolerant than in non-tolerant rats.(ABSTRACT TRUNCATED AT 250 WORDS)


Intracerebroventricular injection of insulin or glucose alters insulin-like growth factor II (IGF-II) concentrations in specific hypothalamic nuclei. Peripherally administered insulin has been shown to alter content and gene expression of hypothalamic insulin-like growth factor II (IGF-II) in a region specific manner (Lauterio, TJ. et al., Endocrinology, 126 (1990) 392-398. The objective of this experiment was to determine whether central administration of insulin can modulate hypothalamic IGF-II peptide content. Male Sprague-Dawley rats were implanted with lateral ventricular cannulae and allowed to recover from surgery one week prior to injection. At that point animals were remotely injected with one of the following: (1) synthetic cerebral spinal fluid vehicle (sCSF); (2) 2 mIU porcine insulin (I); (3) glucose (100 mg%) only. Animals were decapitated 30 min after injections and brains were quickly removed, frozen and dissected into specific hypothalamic regions for IGF-II analysis by RIA. Insulin increased IGF-II content in the ventromedial hypothalamic region by 80% (P < 0.001) and paraventricular nucleus by 30% (P < 0.01) compared to sCSF or glucose treatment. Arcuate nucleus and neurointermediary lobe pituitary IGF-II content was decreased with insulin treatment compared to controls (P < 0.01). Insulin had no effect on IGF-II concentrations in the dorsomedial or lateral hypothalamic regions or in the supraoptic and suprachiasmatic nuclei. Peripheral concentrations of glucose, insulin and IGF-II were unaffected by any treatment. Results show that insulin which reaches the brain can alter IGF-II levels in specific regions of the hypothalamus and suggests a possible role for IGF-II in insulin mediated changes in metabolism or hypothalamic hormone secretion.


Oxidative stress induced by administration of the neuroleptic drug haloperidol is attenuated by higher doses of haloperidol. The effect of haloperidol administration on lipid peroxidation and glutathione/protein thiol homeostasis in the brain was examined 4 h following subcutaneous administration of a single dose of haloperidol; 1.0, 1.5, 2.0 or 2.5 mg/kg b.wt. Glutathione (GSH) levels decreased significantly in cortex, striatum and midbrain after haloperidol administration. Maximal decrease of GSH was observed in the striatum. The depleted GSH was recoverable as protein glutathione mixed disulfide (Pr-SSG) with concomitant loss of protein thiols (Pr-SH) in all the regions of the brain examined. Administration of 1.5 mg/kg b.wt. of haloperidol resulted in significant depletion of GSH in striatum and midbrain as compared to that after administration of the lower dose of 1.0 mg/kg b.wt. of haloperidol. However, administration of higher doses of haloperidol (2.0 and 2.5 mg/kg b.wt.) did not result in greater depletion of GSH; the GSH levels were not significantly different from that observed following the administration of 1.5 mg/kg b.wt. of haloperidol. However, Pr-SSG levels increased dose-dependently following haloperidol administration. The total GSH recovered as sum of GSH and Pr-SSG was significantly higher than controls in striatum and midbrain following administration of higher doses of haloperidol, namely, 2.0 and 2.5 mg/kg b.wt. The depleted GSH was not recoverable as glutathione disulfide (GSSG). GSSG levels were not significantly different from controls 4 h after administration of 1.5 mg/kg b.wt. of haloperidol. The levels of malondialdehyde (indicative of lipid peroxidation) increased significantly as compared to control levels (280-220%) following administration of 1.0 and 1.5 mg/kg b.wt. of haloperidol.(ABSTRACT TRUNCATED AT 250 WORDS)


Constitutive expression of 65-kDa heat shock protein (HSP65)-like immunoreactivity in cultured mouse oligodendrocytes. The expression of mycobacterial 65-kDa heat shock protein (HSP65)-like immunoreactivity in cultured mouse oligodendrocytes and astrocytes was investigated using three monoclonal antibodies (ML30, IA1, 3A) specific for the mycobacterial HSP65. In western blot analysis, these antibodies recognized the proteins with molecular weights approximately of 50-, 60-, and 70-kDa expressed in both heat-stressed and unstressed glial cells. When the cells were exposed to heat stress, the expression of both 50- and 70-kDa proteins was attenuated, whereas that of the 60-kDa protein was not affected. On immunocytochemical studies, an appreciable level of HSP65 immunolabelling was identified in most (> 90%) oligodendrocytes under both heat-stressed and unstressed conditions but only marginally detectable in most (> 95%) astrocytes. These results indicate that mouse oligodendrocytes in vitro express the mycobacterial HSP65-like immunoreactivity constitutively.


The origin of rhythmic fast subthreshold depolarizations in thalamic relay cells of rats under urethane anaesthesia. Intracellular recordings were performed in relay neurons of the dorsal thalamus in rats under urethane anaesthesia. In 77 out of 127 neurons of the ventro-posterolateral and ventral lateral nuclei, but not in neurons of the ventro-posteromedial and posterior nuclei, a highly rhythmic pattern of subthreshold depolarizations was present at rest. The average frequency of these rhythmic depolarizations in ventro-posterolateral cells was 23.36 +/- 11.48 Hz (range: 6-60 Hz); in ventral lateral relay cells higher frequencies were observed (65.86 +/- 17.42 Hz; range: 17-95 Hz). The rhythmic subthreshold events were identified as excitatory postsynaptic potentials generated by the regular firing of prethalamic afferents located in dorsal column and deep cerebellar nuclei. Indeed, in cells of the ventro-posterolateral nucleus these spontaneous potentials had a waveform similar to that of synaptic potentials triggered by somatosensory stimulation. They increased in amplitude with membrane hyperpolarization and their rhythmic occurrence was not affected by the injection of large inward currents. Moreover, they persisted after capsular transection, but they could no more be recorded in ventro-posterolateral cells after lesion of dorsal column nuclei. Finally, it was found that prethalamic afferents within the deep cerebellar nuclei discharged spontaneously in a rhythmic manner within the same frequency band as that of the rhythmic synaptic potentials recorded in ventral lateral cells. On the basis of these results, it is concluded that the rhythmic subthreshold depolarizations observed in thalamic neurons of animals under urethane anaesthesia are not generated intrinsically but that they represent excitatory postsynaptic potentials of prethalamic origin.(ABSTRACT TRUNCATED AT 250 WORDS)


The intrastriatal 6-hydroxydopamine model of hemiparkinsonism: quantitative receptor autoradiographic evidence of correlation between circling behavior and presynaptic as well as postsynaptic nigrostriatal markers in the rat. Unilateral injections of 6-hydroxydopamine into the striatum resulted in almost immediate ipsilateral amphetamine (AMPH)- and delayed contralateral apomorphine (APO)-induced circling behavior in rats. APO-induced rotation correlated positively with that caused by AMPH. In these animals, there was an almost complete disappearance of dopamine uptake sites as well as increases in DA D2 receptors in specific subdivisions of the ipsilateral caudate-putamen (CPu). Both the rate of AMPH- and APO-induced rotation correlated with the percentage of DA terminal loss in the total aspect and in various quadrants of the striatum. In contrast, AMPH- and APO-induced rotation correlated with the percentage increase in striatal D2 receptors only in the dorsolateral (DL) aspect of the CPu. These results indicate that both AMPH- and APO-induced rotation can be used to determine the extent of DA terminal loss in the rat basal ganglia. The positive correlation of circling behavior to only changes in DA D2 receptors observed in the DL striatal subdivision provides further evidence for the heterogeneity of the basal ganglia. This model of hemiparkinsonism in the rat which uses a distant intrastriatal approach to the destruction of nigral DA cell bodies may be a more appropriate model to study the regenerative properties of the nigrostriatal DA system. This approach could also be used to more specifically localize peptidergic receptors on midbrain dopamine cell bodies.


Ultrastructural studies of the primate parabigeminal nucleus: electron microscopic autoradiographic analysis of the tectoparabigeminal projection in Galago crassicaudatus. The normal ultrastructure of the parabigeminal nucleus and the morphology and synaptic relationships of tectoparabigeminal terminals have been examined. Five different morphological types of terminals have been observed within the parabigeminal nucleus. Three of these profiles contain round vesicles and make asymmetrical synapses, while two contain pleomorphic vesicles and make symmetrical synapses. Electron microscopic autoradiographic data indicate that labeled tectoparabigeminal terminals represent only one of the three profiles containing round vesicles. Such terminals are primarily presynaptic to dendritic shafts, and several labeled profiles have been observed presynaptic to the same dendrite.


Serotonergic modulation of junctional conductance in an identified pair of neurons in the mollusc Lymnaea stagnalis. Serotonin (5-HT) is shown to modulate electrotonic coupling between two giant peptidergic neurons in the CNS of Lymnaea stagnalis. The primary effect of 5-HT appears to be a rapid and reversible decrease in gap junctional conductance.


Involvement of the subnucleus reticularis dorsalis in diffuse noxious inhibitory controls in the rat. Several lines of evidence have suggested the participation of the caudal medulla, including the subnucleus reticularis dorsalis (SRD), in the supraspinally mediated diffuse noxious inhibitory controls (DNICs). To test this hypothesis directly, DNICs acting on spinal convergent neurones were compared in sham-operated rats and rats with unilateral quinolinic acid-induced lesions of the SRD. Inhibitions produced by heterotopic noxious stimuli (i.e. DNIC), of the C-fibre-evoked responses of the convergent neurones were significantly reduced in the lesioned animals. This depression of DNIC, by 40-45%, was similar no matter where the conditioning stimuli were applied and was not significantly different for neurones recorded ipsilaterally to the lesion from those recorded contralaterally. These results suggest that the SRD is one of the supraspinal relay(s) in the circuitry underlying DNIC in the rat.


A single meal elicits regional changes in bombesin-like peptide levels in the gut and brain. Several studies have demonstrated that exogenous bombesin (BN) elicits a potent satiety effect when administered centrally or systemically. It has been suggested that BN-like peptides may play a physiological role in the control of food intake. The objective of the present study was to determine whether the levels of endogenous BN-like peptides change in response to a meal. All rats were food deprived overnight for a 12 h period. Half the animals were then allowed to feed for 35 min (postprandial group) and the remainder formed the preprandial group. Regions of the brain (hypothalamus, cerebellum, medulla, pons, neocortex, hippocampus, olfactory bulbs, striatum, midbrain and pituitary), gut (oesophagus, fundus, antrum, duodenum, jejunum, ileum, colon) and adrenal glands were analyzed for BN-like peptide levels using radioimmunoassay. Our results indicate significant increases in the levels of BN-like peptides in the hypothalamus and hippocampus, as well as in the antrum of the stomach, after food ingestion. These results are the first to demonstrate the activation of endogenous BN-like peptide mechanisms in response to ingestion. These rapid alterations in peptide levels may support the contention that BN-like peptides play a physiological role in the regulation of ingestive behavior.


Taurine, GABA and GFAP immunoreactivity in the developing and adult rat optic nerve. The localizations of taurine, gamma-aminobutyric acid (GABA) and glial fibrillary acidic protein (GFAP) within the developing rat optic nerve were determined using immunocytochemical techniques on tissues from animals ranging in age from embryonic day 20 to postnatal 28 days. Mature nerves from 3-4-month-old adults were also examined. At the younger ages, taurine immunoreactivity was intense and localized specifically to the optic nerve axons, but by postnatal day 15 and thereafter its predominant localization was in macroglia. Some of these glia were astrocytes as indicated by the specific marker, GFAP. GABA immunoreactivity was present at the same time as taurine but was found only in macroglia. In mature nerves the patterns of taurine, GABA and GFAP distribution (within glia) were highly similar.


Single K(+)-channel currents under steady-state potential conditions in small hippocampal neurons. Small cultured hippocampal neurons from rat embryos were studied with the patch-clamp technique. Single-channel currents from outside-out membrane patches were recorded under steady-state potential conditions. The most frequently found channel types were selective to K+ and showed conductances of about 30 and 80 pS in the range -20 to 0 mV. Two basic kinetic patterns were observed for the 80 pS channels. In one type, the fraction of time spent in open state increased with potential, and in the other type it decreased. For both types of 80 pS channel, the distribution of dwell times in the open state was well described by the sum of two exponentials while three exponentials sometimes were required for dwell times in the closed state. The time constants of the fitted exponentials could vary considerably during an experiment.


Characterization of muscarinic receptor subtypes in Alzheimer and control brain cortices by selective muscarinic antagonists. Subtypes of muscarinic receptors were characterized in the frontal cortices of control and Alzheimer brains, with labelled quinuclidinyl benzilate [3H]QNB and the unlabelled muscarinic antagonists pirenzepine, AF-DX 116, hexahydro-sila-diphenidol (HHSiD), para-fluoro-hexahydro-sila-diphenidol (p-F-HHSiD) and himbacine. High and low affinity sites were observed for both pirenzepine and AF-DX 116 in human control frontal cortices. The majority (76%) of the pirenzepine binding sites showed high affinity to the muscarinic receptors (M1), while the rest of the binding sites had an affinity that was 40 times less. AF-DX 116 displayed two sets of binding sites where the high affinity AF-DX 116 (M2) sites constituted 27%, while the low affinity AF-DX 116 (non-M2 site) was 73%. A single class of binding sites was observed for HHSiD, p-F-HHSiD and himbacine in human frontal cortices. HHSiD showed an affinity in the frontal cortices that was comparable to that of the pirenzepine high affinity binding (M1) sites. The affinity of p-F-HHSiD was three times lower than that of HHSiD but similar to himbacine. A significant increase in the affinity (+ 40%) as well as in the Bmax (+ 99%) value was observed for the pirenzepine high affinity binding sites (M1) in the frontal cortices of Alzheimer brains compared to controls. Similarly, a significant increase was observed in the Bmax value (+ 60%) for the AF-DX 116 low affinity binding sites (non-M2), while no change was found for the high affinity binding sites (M2).(ABSTRACT TRUNCATED AT 250 WORDS)


Neurotoxic effects of the intrastriatal injection of spermine and spermidine: lack of involvement of NMDA receptors. The injection into the rat striatum of the polyamines spermine and spermidine (30-300 nmol) produced, 1 week after injection, a dose related loss of the neuronal markers glutamate decarboxylase and choline acetyltransferase and a decrease in the density of N-methyl-D-aspartate (NMDA) receptors (as labelled with [3H]TCP). In parallel, an increase in peripheral type benzodiazepine (p) binding site density (a marker of the associated glial reaction and macrophage invasion) was observed. Intrastriatal injection of putrescine (300 nmol) did not significantly alter any of these markers. The effect of spermine on these neuronal and glial markers was maximal 3 days after injection, and tended towards control levels at 16 days post injection. The neurotoxic effects of spermine were confirmed by histological analysis demonstrating a massive neuronal loss around the injection site and an accumulation of astrocytes and phagocytes. The neurotoxic effects of spermine (250 nmol) were not antagonised by the previous administration of the NMDA receptor antagonist MK-801 (10 mg/kg, i.p.). Thus polyamine neurotoxicity in vivo does not seem to involve NMDA receptor activation, although it may possibly be related to the multiple effects of these compounds on diverse calcium channels and processes regulating calcium homoeostasis.


Light and electron microscopic localization of alpha subunits of GTP-binding proteins, G(o) and Gi, in the cerebral cortex and hippocampus of rat brain. Antibodies that recognize alpha subunits of G(o), Gi2 and Gi3 were used to evaluate their association with synaptic junctions. G(o), but not Gi, was concentrated within perikaryal and dendritic cytoplasm of a small population of bipolar neurons. All three G-proteins were associated with the intracellular surface of dendritic, axonal and astrocytic plasma membranes and postsynaptic densities (PSDs). However, association with PSDs was more prevalent for the two Gi's than for G(o) while the association with terminals forming putatively excitatory synapses was more prevalent for G(o) and Gi3 than for Gi2. Thus, neuromodulators may modulate the release of excitatory transmitters via activation of presynaptic Gi3 and G(o) and also regulate the opening of Ca2+ and/or K+ channels via activation of Gi's and G(o) at PSDs.


Cultured astrocytes express mRNA for peptidylglycine-alpha-amidating monooxygenase, a neuropeptide processing enzyme. Cultured astrocytes have been previously found to express several neuropeptides, as well as the neuropeptide processing enzyme carboxypeptidase E (CPE). To investigate whether cultured astrocytes contain additional peptide-processing enzymes, Northern blots were screened for peptidylglycine-alpha-amidating monooxygenase (PAM) mRNA. PAM is involved with the formation of amide groups on the C-terminus of numerous peptide hormones and neurotransmitters. Primary cultures of astrocytes contain moderate levels of PAM mRNA, as determined by Northern blot analysis. The level of PAM mRNA in cultured hypothalamic astrocytes is similar to the level expressed in cultured hypothalamic neurons. The relative abundance of PAM mRNA differs up to 6-fold between astrocytes cultured from various brain regions. Astrocytes cultured from hypothalamus have high levels of PAM mRNA, those cultured from striatum, frontal cortex, and hippocampus have moderate levels, and those cultured from cerebellum have low levels. To investigate whether all cultured astrocytes express PAM mRNA, in situ hybridization analysis of cultured astrocytes was performed. Interestingly, virtually all of the astrocytes cultured from either hypothalamus or cerebellum express PAM mRNA, in contrast to a previous finding that only 20-40% of similarly cultured astrocytes express CPE. The presence of PAM mRNA in cultured astrocytes suggests that these cells have the capacity to produce amidated neuropeptides.


Role of central ATP-sensitive potassium channels in the analgesic effect and spinal noradrenaline turnover-enhancing effect of intracerebroventricularly injected morphine in mice. Glibenclamide is one of the most potent sulfonylurea-derived antidiabetic drugs which block the adenosine triphosphate-sensitive potassium (KATP) channels. In the present study, we found that none of morphine, U-50,488H (a selective kappa agonist) and baclofen (a selective GABAB agonist) added to the incubation medium at concentrations up to 10(-4) M had appreciable effect on the specific binding of [cyclohexyl-2,3-3H(N)]glibenclamide ([3H]glibenclamide) to the isolated mouse brain microsomes. The analgesic activity induced by intracerebroventricular injection (i.c.v.) of morphine but not U-50,488H was antagonized by pretreatment with either i.c.v. glibenclamide or beta-funaltrexamine (beta-FNA; a selective mu antagonist) in mice. Furthermore, the increasing effect of i.c.v. morphine on the spinal noradrenaline (NA) turnover was greatly antagonized by i.c.v. pretreatment with either beta-FNA or glibenclamide. From these results, we demonstrated that KATP channels play an important role as indirect modulators of the supraspinal analgesia induced by mu agonist but not kappa agonist in mice, and the activation of descending noradrenergic system induced by i.c.v. morphine appears to be suppressed by the blockade of KATP channels.


Immunohistochemical localization of calretinin in the rat lateral geniculate nucleus and its retino-geniculate projection. In the present study, we examined the distribution of calretinin-immunoreactive neuronal cell bodies and fibers in the lateral geniculate nucleus of the rat. In normal rats, clusters of immunoreactive cell bodies were found in: (i) the rostral portion of the ventral lateral geniculate nucleus pars medialis (VLGM), (ii) the intergeniculate leaflet (IGL), (iii) the intermediate region between the VLGM and the ventral lateral geniculate nucleus pars lateralis (VLGL), (iv) the caudomedial portion of the VLGM, and (v) the caudolateral portion of the VLGM. In the dorsal lateral geniculate nucleus (DLG), immunoreactive cell bodies were rarely observed. After uni- or bilateral eye enucleation, no significant alteration in the morphological features or distribution of immunoreactive cell bodies was detected in the lateral geniculate nucleus. In normal rats, immunoreactive fibers formed dense plexuses in: (i) the DLG, (ii) the external layer of the VLGL, (iii) the internal layer of the VLGL, (iv) the IGL, (v) the caudomedial portion of the VLGM, and (vi) the optic tract. After unilateral eye enucleation, immunoreactive fibers in the external layer of the VLG and in the optic tract almost totally disappeared on the contralateral side to the lesion. Unilateral eye enucleation caused a significant decrease of immunoreactive fibers in the DLG and in the internal layer of the VLGL, but a substantial number of immunoreactive fibers still remained there. In the IGL and the caudomedial portion of the VLGM, no observable alteration in the distribution of immunoreactive fibers was detected after uni- or bilateral eye enucleation.(ABSTRACT TRUNCATED AT 250 WORDS)


Acute depolarization block of A10 dopamine neurons: interactions of morphine with dopamine antagonists. Extracellular single-unit recording techniques were used to determine the effects of morphine, administered either systematically (intravenous) or locally (microiontophoresis), on ventral tegmental area (A10) dopamine (DA) neuronal activity in animals pretreated with D1 (SCH 23390) or D2 (pimozide) DA receptor antagonists. In rats pretreated with the D2 antagonist pimozide, A10 DA neurons readily entered a state of apparent depolarization block in response to either i.v. or iontophoretically applied morphine. Whether the inactivation of DA neurons was induced by systemic or local morphine, it was reversed in 22 of 27 cases by iontophoretic administration of the inhibitory amino acid transmitter gamma-aminobutyric acid (GABA), suggesting depolarization block as the underlying mechanism. Pretreatment of rats with the D1 antagonist SCH 23390 did not significantly alter the tendency of A10 DA neurons to enter apparent depolarization block in response to morphine. These data support recent behavioral evidence suggesting that the combination of systemic pimozide and ventral tegmental area morphine can result in depolarization inactivation of the mesoaccumbens DA reward system.


Chronological expression of microtubule-associated proteins (MAPs) in EC cell P19 after neuronal induction by retinoic acid. Pluripotent murine embryonal carcinoma (EC) P19 cells are induced at a high rate into neural cells using retinoic acid and serum-free medium. EM observation revealed great increase of microtubules (MTs) after neuronal induction. To study the expression of microtubule-associated proteins (MAPs), immunoblotting and immunocytochemistry were performed with phosphorylated MAP1B (pMAP1B)-, MAP2-, and MAP1A-specific monoclonal antibodies. They did not stain undifferentiated cells. Early MAPs (pMAP1B and MAP2C) appeared 12 h after the neuronal induction, changing to late MAPs (MAP1A and MAP2A/B) at 3-5 days. These expression patterns are quite similar to those of neural cells in vivo. Anti-pMAP1B stained not only neurites but also the cell body and varicosities. But after extraction of the soluble component by permeabilization, pMAP1B was found in only MT-domains of the neurites at LM and EM levels, indicating that some part of pMAP1B is a structural component of neurite MTs and others exist in a soluble form. After culturing for more than 5 days, pMAP1B disappeared from the soma, but still remained in the distal ends of neurites. Here we showed that P19 is a good model system for studying the expression of MAPs on the continuous course of neuronal differentiation.


Biologically active salmon calcitonin-like peptide is present in rat brain. Although high densities of calcitonin (CT) receptors and potent CT-induced actions occur within the rat central nervous system, the physiological ligand for central receptors remains unidentified. This study identifies a salmon CT (sCT)-like peptide in extracts of rat brain. Aliquots of 1 M HCl brain extracts were reconstituted in 0.01 M acetic acid and assayed for sCT-like immunoreactivity using a radioimmunoassay (RIA) based on a guinea pig anti-sCT antibody (detection limit 0.1 ng). The assay was less than 0.1% cross-reactive with human CT, porcine CT, rat calcitonin gene-related peptide (CGRP)-I and human CGRP-I. Extracts of diencephalon revealed sCT-like activity (30-120 ng/g wet weight), while extracts from cortex, cerebellum and medulla oblongata did not contain detectable activity. Reverse-phase HPLC analysis demonstrated co-elution with synthetic sCT. Further, the peak material was active in 4 CT bioassay systems; (1) stimulation of cAMP production in UMR 106-06 cells and (2) T47D cells, (3) inhibition of 125I-sCT binding in UMR 106-06 cells and (4) inhibition of 125I-sCT binding in sheep brain membranes. The bioactivity of material in these assays was similar to that estimated through the RIA. The results identify, for the first time, the presence of sCT-like material in the rat brain and constitutes the first demonstration of biologically active sCT-like material from brain extracts. Salmon CT-like peptide may represent the endogenous ligand for central CT receptors.


The time course of sigma activity and slow-wave activity during NREMS in cortical and thalamic EEG of the cat during baseline and after 12 hours of wakefulness. The extrapolation from recent neurophysiological findings concerning the dependency of spindle and slow-wave oscillations of thalamocortical neurons on membrane potential to macroscopic EEG events, predicts a reciprocal relation between spindle activity and slow-wave activity (SWA) in thalamic and cortical EEG during non-rapid-eye-movement sleep (NREMS). To test this hypothesis, the EEG recorded in 8 cats, from the nucleus centralis lateralis of the thalamus and from the skull during a 12-h baseline dark period and during a 12-h recovery dark period, following a 12-h sleep deprivation, were analyzed. Per 12-s epoch, sleep-wake behaviour was determined and spectral power density was computed in the slow-wave frequency range (0.5-4.0 Hz) and in the spindle frequency region (sigma activity: 11.0-14.5 Hz). To analyze the development of EEG power densities in the course of NREMS and during the transition from NREMS to REMS, the last epoch of wakefulness and the first 15 epochs of NREMS, as well as the last epochs of NREMS and the first epoch of REMS were selected from the NREM-REM cycles. For each animal the values were averaged over 4-h intervals. In the cortical EEG, SWA was minimal at NREMS onset and increased progressively in the course of NREMS. SWA declined sharply prior to REMS. sigma Activity increased gradually towards a uniform level after NREMS onset. During the transition to REMS, sigma activity initially increased and then decreased rapidly. In the thalamic EEG, the time course of SWA paralleled that of the cortex. However, the development of sigma activity during the first part of NREMS differed: in the thalamic EEG, sigma activity was maximal during the beginning of NREMS and slightly decreased thereafter. After sleep deprivation, SWA within NREMS was markedly enhanced in both the cortical and the thalamic EEG. Sigma activity was attenuated in the thalamic EEG, whereas in the cortical EEG it was temporarily elevated. The present data show that, in the thalamic EEG, an inverse relation exists between spindle and slow-wave activity during baseline NREMS. This relation is preserved after sleep deprivation. In the cortical EEG, a reciprocal relation between spindling and SWA is less evident.


Peroxidative stress effects on calpain activity in brain of young and adult rats. Three hours after administration of the pro-oxidant 2-cyclohexen-1-one, calpain activity was significantly reduced in the brain of young rats, but not in the brain of adult rats, and cathepsin D activity remained unchanged. Addition of isovalerylcarnitine to the incubation medium increased calpain activity 5-7-fold, counteracting the effect of the pro-oxidant.


Parasympathetic postganglionic nerve fibers in the fungiform papillae of the bullfrog, Rana catesbeiana. An investigation was made of the precise origin of the unmyelinated nerve fibers in the fungiform papillae of the bullfrog's tongue. Some unmyelinated nerve fibers in the fungiform papillae originate from the parasympathetic postganglionic cells in the glossopharyngeal nerve. Axonal enlargements of the parasympathetic nerve fibers were in close contact with the Merkel-like basal or supporting cells in the taste disk. These results seem to provide morphological evidence for the existence of an efferent control system in the taste disk.


Zinc modulates GABAB binding in rat brain. The effects of ZnCl2 on [3H]GABA binding to GABAA and GABAA binding sites were investigated using receptor autoradiography. At concentrations exceeding 100 microM, zinc non-competitively inhibited GABAB binding in a dose dependent fashion. GABAA binding was not inhibited significantly by zinc eliminating the possibility of a non-specific effect of zinc. Increased calcium concentrations up to 10 mM enhanced total GABAB binding but did not prevent zinc induced inhibition of GABAB binding, indicating a separate site of action for these cations at the GABAB binding site. In some regions, zinc modulates GABAB binding in a biphasic manner as concentrations of 10-100 microM zinc significantly enhanced GABAB binding in the hippocampus and the molecular layer of the cerebellum but not in the thalamus. These results provide further evidence for a neuromodulatory role for zinc in the central nervous system.


Observations on morphology and electrophysiological properties of the normal and axotomized facial motoneurons in the cat. The correlation between the morphology of facial motoneurons stained intracellularly with horseradish peroxidase and their physiological parameters was examined in cats following facial nerve section and in cats with intact facial nerve. A certain statistical relationship exists between cell size and excitability in normal neurons. After axotomy, facial neurons showed a slow conduction velocity and a low rheobasic current, but had a normal cell size. Physiological changes include repetitive firing in response to intracellular current injection, reflecting an increase in the excitability in the axotomized neurons.


Sex differences in the regulation of heat shock protein 70 kDa and 90 kDa in the rat ventromedial hypothalamus by estrogen. The regulation of heat shock proteins 70 kDa (hsp 70) and 90 kDa (hsp90) by estradiol (EB) was examined in the ventromedial hypothalamus (VMH) of gonadectomized adult female and male rats with and without estradiol replacement. Estradiol significantly elevated hsp70 and hsp90 protein concentrations in EB-treated VMH of females, but not males, sacrificed at 12 h following steroid hormone compared with oil-vehicle controls. These findings confirm our previous observations that EB induces hsp70 and hsp90 in the female VMH16,17. The sex differences for the two proteins may occur by different mechanisms, since basal hsp70 levels were higher in males than females, whereas hsp90 levels were similar for the two sexes.


Repeated focal cerebral ischemia in gerbils is associated with development of infarction. We induced repeated focal cerebral ischemia in gerbils. Single 5-min ischemia produced neuronal damage limited to the ipsilateral CA1 and CA4 hippocampus. Two 5-min ischemic insults spaced at a 1-h interval caused selective neuronal damage to the CA1, CA3 and CA4 hippocampus, striatum, neocortex, and thalamus. Three 5-min ischemic insults at 1-h intervals produced infarction. Thus, repeated focal ischemia produced cumulative brain damage by conversion of sublethal damage into selective neuronal damage and of the neuronal damage into infarction.


Lack of neuroprotective effect of some sigma ligands in a model of focal cerebral ischemia in the mouse. The potential neuroprotective effect of seven sigma ligands has been evaluated in a mouse model of focal cerebral ischemia (induced by coagulation of the left middle cerebral artery) and compared to that of known N-methyl-D-aspartate (NMDA) antagonists. When given after the induction of cerebral ischemia, the NMDA receptor antagonists dizocilpine and CGS 19755 and the mixed NMDA antagonist/sigma ligand eliprodil (SL 82.0715) afforded very substantial protection against cortical infarction (92, 44 and 72%, at the doses of 1, 10 and 10 mg/kg, i.p., for dizocilpine, CGS 19755 and eliprodil, respectively). In contrast, none of the sigma ligands investigated--DTG, DMTG, GBR 12909, (+)- and (-)-3-PPP (up to 10 mg/kg), BMY 14802 (up to 30 mg/kg), except haloperidol at a high dose (3 mg/kg)--had neuroprotective effects.


Enhanced gastric acid secretion induced by gastrin can be suppressed partially by glucose injection into the medullary nuclei in rats. Gastric acid outputs after glucose injection into the nucleus of the vagus nerve (X) or into the nucleus of the tractus solitarius (SOL) were examined in rats with tetragastrin. The enhanced acid output caused by gastrin was partially inhibited by glucose injection into the X or SOL. Glucose injection into the X depressed acid output in a dose-dependent fashion, but a non-dose-dependent response in acid output was seen when glucose was injected into the SOL. There was no interaction between the X and SOL in acid response due to glucose. These findings suggest that glucose modulates gastric acid secretion stimulated by gastrin at the medullary level, and that such a modulation is characterized by the medullary X or SOL, separately.


Differential laminar distribution of acetylcholinesterase and butyrylcholinesterase containing tangles in the cerebral cortex of Alzheimer's disease. A sensitive histochemical method for the visualization of acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) activity was used to determine the laminar distribution of cholinesterase-positive cortical tangles in Alzheimer's disease (AD). In many cortical areas AChE- and BChE-positive tangles displayed a completely overlapping distribution. In other areas the most superficial layers contained only AChE-positive tangles whereas the deepest layers contained only BChE-positive tangles. These observations suggest that some cholinesterase-positive tangles have a predominantly (if not exclusively) AChE-like reactivity whereas others have a reactivity that is predominantly BChE-like. The intermingling of AChE- and BChE-positive tangles in most cortical areas and layers suggests that there may also be a third population in which the two enzymes are equally prominent in the same tangle.


Genetically variable taste sensitivity to D-amino acids in mice. Behavioral and neural responses to D-amino acids were compared between two inbred strains, C57BL and BALB mice. In both strains, an aversion conditioned to D-valine, D-leucine, D-methionine, D-histidine or D-tryptophan generalized to sucrose, whereas an aversion to D-alanine or D-serine did not generalize to sucrose. Generalization patterns across various test stimuli for each of these 7 D-amino acids were significantly correlated between two strains. However, an aversion conditioned to D-phenylalanine generalized to sucrose in C57BL mice, but not in BALB mice. Application of a proteolytic enzyme, Pronase E, to the tongue reduced chorda tympani responses to sucrose and D-amino acids to which a conditioned aversion generalized to sucrose. Again, only in C57BL mice, Pronase inhibited D-phenylalanine responses. These comparable results indicate that sweet taste response is genetically highly variable only to D-phenylalanine among 8 D-amino acids tested.


Vasopressinergic modulation of stress responses in the central amygdala of the Roman high-avoidance and low-avoidance rat. The central nucleus of the amygdala (CEA) is selectively involved in the passive component of the behavioral (immobility) and the accompanying parasympathetic response during conditioned, stressful environmental challenges. Vasopressinergic mechanisms in the brain seem to play a role in these stress responses. The effects of the neuropeptides arginine-8-vasopressin (AVP) and oxytocin (OXT) on modulating CEA activity during conditioned stress of inescapable footshock were studied in male Roman high-avoidance (RHA/Verh) and low-avoidance (RLA/Verh) rats, psychogenetically selected on the basis of shuttle-box acquisition behavior. In RLA/Verh rats, the cardiac and behavioral responses to the conditioned emotional stressor were bradycardia and immobility, suggesting an important role for the CEA in these rats. The RHA/Verh rats, however, failed to show any change in heart rate or immobility in response to a conditioned stress situation. The low dose of AVP (20 pg) in the CEA of conscious RLA/Verh rats caused an enhancement of the stress-induced bradycardiac and immobility response. However, the high dose of AVP (2 ng) and OXT (200 pg) attenuated the bradycardiac and immobility responses in the RLA/Verh rats. Infusion of AVP and OXT in the RHA/Verh rats failed to induce any change in heart rate or immobility. Binding studies revealed that the AVP receptor selectively binds AVP with high affinity. In contrast, the OXT receptor recognizes both AVP and OXT with a similar (but lower) affinity. This suggests that the behavioral and autonomic responses of the high dose of AVP may be caused by OXT receptor stimulation.(ABSTRACT TRUNCATED AT 250 WORDS)


Estrous cycle variation of afferent fibers supplying reproductive organs in the female rat. Multi-unit afferent nerve activity was recorded from branches of the hypogastric and pelvic nerves in virgin female rats on different days of the estrous cycle. In each rat, the response of hypogastric nerve fibers to uterine distension and the response of pelvic nerve fibers to vaginal distension was tested. The minimal pressure necessary to evoke a response was highest in diestrus for both the hypogastric and pelvic nerve fibers. For the hypogastric nerve, the minimal necessary pressures were significantly lower during both proestrus and estrus, whereas for the pelvic nerve, the pressure was significantly lower only on the day of proestrus. These results suggest that the overall response sensitivity of afferent fibers in the pelvic and hypogastric nerves are differentially affected by hormonal variations occurring across the estrous cycle in a manner that would enhance reproduction.


Colocalization of enkephalin-, glucagon-, and corticotropin-releasing factor-like immunoreactivity in GABAergic amacrine cells in turtle retina. The large number of amacrine cells which contain gamma-aminobutyric acid (GABA) in the turtle retina makes it difficult to examine specific GABAergic cell types. In order to selectively label subpopulations of GABAergic neurons, we have used fluorescent double-labeling immunocytochemical techniques to examine the localization of GABA-like immunoreactivity (LI) in amacrine cells which contain antigens resembling the neuropeptides glucagon (GLUC), corticotropin-releasing factor (CRF) or enkephalin (ENK). GABA-LI was found in 41% of the cells with GLUC-, 100% of the cells with CRF-, and 69% of the cells with ENK-LI. There were regional differences in the presence of GABA-LI in amacrine cell populations with ENK-LI. GABA-LI was present in about 80% of the cells with ENK-LI outside of the visual streak, while only 37% of the cells within the streak had GABA-LI. Based on the distinct morphologies and regional distributions of these peptidergic amacrine cells, we conclude that they represent different subpopulations of GABAergic amacrine cells in the turtle retina. Future studies can now utilize existing information regarding the synaptic connectivity of these peptidergic amacrine cells to help delineate the functions of GABAergic amacrine cells in the turtle retina.


Vasopressin-induced motor effects: localization of a sensitive site in the amygdala. Arginine vasopressin (AVP) induces motor effects when administered into the cerebral ventricles, the ventral septal area (VSA), or the vestibular cerebellum of the rat brain. Because AVP-like immunoreactivity and AVP-binding sites exist in the central medial amygdala (cmeA), and because the amygdala can be kindled to produce motor effects, we hypothesized that the amygdala might play a role in AVP-induced motor effects. This hypothesis was tested by observing motor behavior in response to injection of AVP into the central medial region of the amygdala. Our results demonstrate that an initial injection of AVP into the cmeA caused minor motor effects, including immobility, prostration and ataxia, whereas a similar injection, given 24 h later, caused severe motor effects including barrel rotations and myoclonic/myotonic-like convulsive behavior. A potential receptor basis for the AVP-induced motor and sensitization effects in the cmeA was investigated using AVP analogues. A V1 antagonist, d(CH2)5Tyr(Me)AVP, blocked both the motor and sensitization effects produced by cmeA AVP injection. A V2 receptor agonist, DDAVP, did not affect motor activity upon cmeA injection, but did, however, sensitize animals to subsequent cmeA AVP injection. These results suggest that the cmeA is a sensitive site for AVP-induced motor effects and that these motor effects are sensitized by prior exposure to AVP. While the motor effects observed after cmeA AVP injection are mediated via AVP receptors that resemble the V1 type, the sensitization effect may be mediated via multiple receptor systems.(ABSTRACT TRUNCATED AT 250 WORDS)


Lesions of the nucleus paragigantocellularis: effects on mating behavior in male rats. We evaluated the effects of bilateral radio-frequency lesions of the paragigantocellular (PGi) reticular nucleus in the ventral medulla on male rat copulatory behavior. In Experiment 1, sexually naive male rats with such lesions were more likely than sham-operated controls to copulate to ejaculation during their first exposure to an estrous female. Additionally, among the rats that copulated to ejaculation, those with lesions demonstrated a reduction in mount frequency (MF), intromission frequency (IF), and ejaculation latency (EL), and an increase in copulatory efficiency (CE). In Expt. 2, sexually experienced male rats were allowed to mate to sexual exhaustion. Males with PGi lesions showed an increased latency to sexual exhaustion and an increased number of ejaculations prior to exhaustion. Additionally, rats with PGi lesions displayed reductions in IF, EL, and post-ejaculatory interval (PEI) as they approached sexual exhaustion. Our results provide further evidence that the PGi is a supraspinal locus of descending inhibitory influence on spinal nuclei mediating ejaculatory reflexes in the male rat.


Androgen and estrogen concentrating neurons in chemosensory pathways of the male Syrian hamster brain. The medial preoptic area (MPOA), bed nucleus of the stria terminalis (BNST), and medial amygdaloid nucleus (Me) are essential for male sexual behavior in the Syrian hamster. These nuclei received chemosensory stimuli and gonadal steroid signals, both of which are required for mating behavior. The objective of this study was to compare the distribution of androgen- and estrogen-concentrating neurons in MPOA, BNST, and Me in the adult male hamster using steroid autoradiography for estradiol (E2), testosterone (T) and dihydrotestosterone (DHT). Adult males (n = 4 per group) received two i.p. injections of tritiated steroid 4-7 days after castration. Six-microns frozen sections through the brain were mounted onto emulsion-coated slides, and exposed for 11-16 months. In MPOA, BNST, and Me, neurons were more abundant and heavily labelled after [3H]E2 treatment than after either [3H]T or [3H]DHT. Tritiated estradiol- and DHT-labeled cells were found throughout the rostrocaudal extent of Me, with a high concentration in posterodorsal Me. Tritiated testosterone treatment labelled cells largely within posterodorsal Me. In MPOA, the majority of E2-, T-, and DHT-labelled neurons were in the medial preoptic nucleus (MPN) and the preoptic continuation of the posteromedial bed nucleus of the stria terminalis (BNSTpm). Few T-labelled cells were present outside these subdivisions. In the BNST, E2- and DHT-labelled neurons were present in all subdivisions, whereas T labelling was confined to the antero- and posteromedial subdivisions of BNST. These results suggest that the distribution of androgen- and estrogen receptor-containing neurons overlap considerably in nuclei which transmit chemosensory signals in the control of mating behavior.


Blockade of NMDA receptor-channels by MK-801 alters breathing in adult rats. The role of N-methyl-D-aspartate (NMDA) receptor-channel activation in the production of respiratory pattern was studied by administration of the NMDA receptor-channel blocker (+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine hydrogen maleate (MK-801, 1-3 mg/kg, i.v.) to anesthetized adult rats. This dose of MK-801 blocked the excitatory effects of NMDA (applied iontophoretically) on brainstem respiratory neurons. The predominant respiratory response to systemic MK-801 administration was an increase in inspiratory duration and a decrease in amplitude of diaphragm electromyogram and phrenic nerve discharge. Effects on inspiratory timing and amplitude were most pronounced when the rats were vagotomized. Significant changes in arterial blood gases and pH after systemic MK-801 administration in spontaneously breathing rats (vagi intact or cut) indicated that ventilation was depressed by NMDA receptor-channel antagonism. Respiratory timing changes in response to systemic MK-801 administration differed between two rat strains studied. Breathing patterns resembling apneusis, i.e., with irregular inspiratory durations prolonged 2- to 30-fold, occurred in 60% of the vagotomized, spontaneously breathing Sprague-Dawley rats and none of the Wistar rats. Thus, the breathing pattern in Sprague-Dawley rats is more sensitive to interference with NMDA-mediated mechanisms. We propose that respiratory pattern generation and transmission of rhythmic respiratory drive are mediated by synergistic activation of NMDA and non-NMDA receptors at brainstem and spinal cord sites.


Fast, statistically based alignment of amino acid sequences on the base of diagonal fragments of DOT-matrices. We present a new pairwise alignment algorithm that uses iterative statistical analysis of homologous subsequences. Apart from the classical conversion of the DOT-matrix characteristic of the Needleman-Wunsch algorithm (NW), we used only those matrix elements that corresponded to the most non-random subsequence homologies. The most reliable elements of the DOT-matrix are written to the compact competition matrices. The algorithm then searches for alignment on the base of only these matrix elements. Our algorithm has low storage and memory requirements, but provides a reliable alignment for the sequences of weak homology (or, at least for the homology regions). In such cases classical NW algorithms often produce unreliable results on the level of statistical noise due to accumulation of random matchings throughout the aligned sequences.


MALIGNED: a multiple sequence alignment editor. A multiple sequence alignment editor is described which runs on a VAX/VMS system and can exchange data with a number of other programs, including those of the Genetics Computer Group (GCG). Up to 199 sequences can be aligned. The quality of the alignment can be easily judged during its development because the display attributes to each character are determined by the way it matches the other sequences. Four methods are available for calculating the highlighting to emphasize different aspects of the relationships of the sequences and up to four styles of highlighting can be used at the same time. Laser printer output is suitable for publication without modification.


Estimation of restriction maps with known site order using a generalized linear model. A generalized linear model with Gamma errors is used to estimate the coordinates of a restriction map when the site order is known. This can be conveniently programmed in a wide range of statistical packages (e.g. Genstat 5, Minitab, SAS), and gives maximum likelihood estimates with their associated optimal properties. Regression diagnostics allow the checking of assumptions and help to identify mis-specified, influential or discordant fragment lengths. A specific diagnostic for identifying fragment lengths causing reversal of restriction site order is derived. Exact 'fragment' lengths from DNA sequencing can be conveniently included in an approximate manner by giving them a larger weight than observed restriction fragment lengths. Two examples and the Genstat 5 codes used in their analysis are presented.


APL program for evaluation of heat processes for spherical foods. The present program solves the heat transfer equations for spherical foods by a numerical procedure that uses the Crank-Nicolson method. The program is implemented for conditions with as few restrictions as possible to enhance versatility. The results agree with the analytical solutions for two particular cases of boundary conditions. The program also computes the thermal effect in the foodstuff from the temperature data obtained by the numerical program.


Cumulative logit modelling for ordinal response variables: applications to biomedical research. Incorrect statistical methods are often used for the analysis of ordinal response data. Such data are frequently summarized into mean scores for comparisons, a fallacious practice because ordinal data are inherently not equidistant. The ubiquitous Pearson chi-square test is invalid because it ignores the ranking of ordinal data. Although some of the non-parametric statistical methods take into account the ordering of ordinal data, these methods do not accommodate statistical adjustment of confounding or assessment of effect modification, two overriding analytic goals in virtually all etiologic inference in biology and medicine. The cumulative logit model is eminently suitable for the analysis of ordinal response data. This multivariate method not only considers the ranked order inherent in ordinal response data, but it also allows adjustment of confounding and assessment of effect modification based on modest sample size. A non-technical account of the cumulative logit model is given and its applications are illustrated by two research examples. The SAS programs for the data analysis of the research examples are available from the author.


GeneScape: a relational database of Escherichia coli genomic map data for Macintosh computers. We present a relational database program developed in FoxBase+/Mac for the viewing and manipulation of ordered restriction maps and associated features of the Escherichia coli genome including sequenced genes and the Kohara miniset of bacteriophage lambda clones. Use of this program allows easy access to the wealth of information being collected in a dataset of DNA sequences, maps and genetic data known as EcoSeq, EcoMap and EcoGene respectively.


Can we recover a sequence, just knowing all its subsequences of given length? The problem tackled here concerns the feasibility of DNA sequencing using hybridization methods. We establish algorithms for and computational limitations to the reconstruction of a sequence from all its subsequences having the same length: in other words, the building of a string that contains all the words of a given set, and only these ones. Generally there are several possible strings. We refer to graph theory and propose an algorithm to enumerate all the strings that are solutions. We then carried out stimulations using real DNA sequences. They provided some necessary conditions and give some upper bounds to the length of the sequence to recover in relation with the length of oligonucleotides. To avoid limiting ourselves to problems that admit a unique solution, we introduce another algorithm that produces a signature for each solution string. Each signature can be tested to determine which one belongs to the correct sequence.


Time correction for computing Michaelis--Menten kinetics. A description of time correction applied to the integrated Michaelis equation in view of cancelling a non-correct estimation of the Michaelis constant due to a lack of coincidence between time zero of measurement and time zero of reaction is presented. An estimation of kinetic parameters is made with time correction on the same experimental set of data using a classical least squares iterative procedure.


AIDS in Ireland: the reporting delay distribution and the implementation of integral equation models. This paper deals with two basic aspects concerning the modelling of AIDS incidence in the context of Irish data. We describe initially the adjustment of the number of AIDS cases (Xij) to allow for reporting delays, where a simple form of the likelihood function for the Xij is supported by GLIM. Subsequently, we consider the accessibility of numerical solution (through a NAG routine) of the integral equation models generated by the back-projection method for the adjusted AIDS cases. Results for the Irish data are summarized for various choices of the incidence distribution.


IBIS integrated biological imaging system: electron micrograph image-processing software running on Unix workstations. 'IBIS' is a set of computer programs concerned with the processing of electron micrographs, with particular emphasis on the requirements for structural analyses of biological macromolecules. The software is written in FORTRAN 77 and runs on Unix workstations. A description of the various functions and the implementation mode is given. Some examples illustrate the user interface.


A program for computer-aided analyses of ecological field data. A program for IBM-compatible microcomputers is introduced which combines several complementary analyses of species-station-tables generated in ecological field investigations. The scope of the program encompasses table editing functions, routines for community delimitation by cluster analysis and procedures for the analysis of properties related both to stations (e.g. diversities) and species (e.g. abundance statistics and association indices). The essential reasoning behind the application of community studies is presented briefly, as well as the multi-step analytical approach implemented in the program.


MacT: Apple Macintosh programs for constructing phylogenetic trees. MacT is a set of programs for the Apple Macintosh to construct and evaluate unrooted trees derived from amino acid sequences using a distance matrix method. Programs are designed on a 'one program--one task' basis for (i) determining the branching order in trees consisting of four or five species and calculating various statistical measures, (ii) calculating statistical measures for all possible topologies of unrooted trees and (iii) generating and evaluating trees derived from bootstrapped samples. With four auxiliary programs unrooted trees can be built for maximal 26 species, and the robustness of topologies be tested by bootstrapping.


A personal computer program for large-scale comparisons of related nucleotide sequences. A personal computer program (COMPSEQ) has been developed which can present an informative listing of pre-aligned exonic nucleotide sequences and of their translations to amino acid sequences as well run triplet-oriented analyses on these sequences in a given reading frame. The sequence listing focuses on the differences between related sequences by suppressing the concordances between them.


OBSTRUCT: a program to obtain largest cliques from a protein sequence set according to structural resolution and sequence similarity. A program OBSTRUCT has been developed to obtain the largest possible subset according to specific constraints from a set of protein sequences whose tertiary structures have been determined crystallographically. The user can request a range in sequence similarity level and/or structural resolution. The program optionally includes sequences with known three-dimensional folds elicited from NMR data.


Isolation and initial characterization of a new virus: Micro-Foci inducing virus or MFV. Infectious and transforming activity was obtained from cultures of a large kidney tumour with very high N-myc amplification. The patient belonged to a cluster of neuroblastoma cases diagnosed in Southern Louisiana in 1986-1988. Infection with ultra-filtered supernatants from neuroblastoma cultures caused the appearance of small foci of rounded cells with neural features and the presence of virus was indicated by electron microscopy: the virus was called Micro-Foci inducing virus or MFV. Infected young adult rats did not develop overt disease, but 11/11 of the litters from infected animals developed a very dramatic and highly lethal syndrome which could resemble neuroblastoma symptoms. N-myc amplification disappeared in very early cultures of the tumour, but it was subsequently detected in MFV-infected cell cultures.


[Remission of established disease in diabetic NOD mice induced by anti-CD3 monoclonal antibody]. Anti-CD3 monoclonal antibodies are potent immunosuppressants widely used in clinical transplantation to prevent or treat acute allograft rejection. We have used a hamster monoclonal antibody (145 2C11) specific for the epsilon chain of the murine CD3 complex to treat autoimmune insulin-dependent diabetes appearing spontaneously by 15 to 30 weeks of age in female Non Obese Diabetic (NOD) mice. Mice showing overt disease (glycosuria and glycemia > or = 4 g/l) were randomized in two groups receiving either anti-CD3 (5 micrograms/day i.v. for 5 consecutive days) or an identical dose of hamster polyclonal immunoglobulins. Progressive remission of disease was observed, 3 to 6 weeks after the end of treatment, in 80% of anti-CD3 treated mice as compared to 6% of mice in the control group. This remission was maintained long term namely, during the 4 to 5 months (after the end of treatment) observation period. These results open interesting perspectives on the possibility to treat recently diagnosed diabetic patients with therapy showing long term efficacy and no chronic toxicity.


[Spiroplasma from Tabanidae capable to multiply and to persist in the central nervous system in mice]. Using alternate passages of cultivation in vitro at 37 degrees C and in suckling mice following intracerebral inoculation, a tabanid spiroplasma was demonstrated capable of multiplication and persistence in mice for 6 successive passages, without production of specific antibody. This is the first report of a spiroplasma from a common flying haematophagous arthropod shown to produce persistent infection of a mammal.


'Horoscope effect' not only for seasonal but also for non-seasonal allergens. We report on the relation between the month of birth and the chance of developing an IgE antibody response as found in a study sample of 150,000 subjects. Our results confirm that for the three seasonal allergens birch pollen, grass pollen and house dust mite, an increased relative risk was found for subjects born up to 3 months before the main season for that allergen in The Netherlands. For cat and dog allergy an increased relative risk was found from November to January, perhaps reflecting increased exposure to these pets during the winter. Surprisingly, however, also for egg white and cow's milk a clearly increased relative risk was found from November to January and a decreased relative risk in May. These data support the hypothesis of a 'sensitive' period in the first months of life during which allergen exposure is more likely to prime for an allergy later in life. The results with the non-seasonal allergens suggest that another seasonal factor exists which early in life assists (or prevents) priming by allergen.


Month of birth and allergic disease at the age of 10. The relationship between month of birth and asthma, hay fever and skin sensitization to mixed grass pollen was analysed in a population-based cross-sectional study in Munich and Bavaria 1989-1990 of 6535 10-year-old children. The relative risk of developing atopic disease is calculated by comparing the prevalence in a single month with the prevalence of all other months. A slightly increased risk of developing allergic skin sensitization for grass pollen (n = 1128) was found for February (odds ratio, 1.3, 95% confidence interval 1.0-1.6), May (1.4, 1.1-1.8) and June (1.3, 1.0-1.6). For hay fever (n = 379) an increase was found for May (1.5, 1.0-2.1) and for allergic asthma (n = 277) for August (1.4, 1.0-2.1). A protective effect was observed for certain months of birth; September for allergic sensitization (0.8, 0.6-1.0), October for and November for hay fever (0.6, 0.3-0.9). The occurrence of hay fever and positive prick test is explained by the seasonal variation of atmospheric grass pollen and the peak in August of asthmatic patients by house dust. Date of birth appears therefore to slightly influence the risk of developing an allergic sensitization and allergic diseases.


Monoclonal antibodies to Lepidoglyphus destructor: delineation of crossreactivity between storage mites and house dust mites. We have developed monoclonal antibodies (MoAbs) to the storage mite Lepidoglyphus destructor (Ld). Employing these anti-Ld MoAbs Ld-MoAbs) in ELISA and ELISA inhibition techniques we have analysed the reaction pattern of Ld-MoAbs to both non-pyroglyphid and pyroglyphid mites. The storage mite Glycyphagus domesticus (Gd) exhibited most efficient inhibition, followed by Acarus siro (As), Tyrophagus putrescentiae (Tp), Dermatophagoides pteronyssinus (Dpt) and Euroglyphus maynei (Em). Of the two pyroglyphid species, Dpt showed at least 1000 times less inhibition than Gd. Two of the MoAbs immunoprecipitated a band of 39 kD whereas the third reacted weakly, with a high-molecular band of approximately 110 kD. The Ld extract was also subjected to various reagents and conditions and the antigen was heat stable, it was not affected by low pH, or sensitive to dimethyl sulphoxide (DMSO) or paraformaldehyde. After exposure of the extract to various reagents, such as protease trypsin and periodate, we conclude that the epitopes recognized by Ld-MoAbs were of carbohydrate rather than of protein nature. It would thus seem that MoAbs recognize the carbohydrate part of a glycoprotein.


Adrenergic response in children with asthma on exogenous stimuli. In asthmatic children it was investigated whether the degree of impairment of the adrenergic response on exogenous stimuli is related to the magnitude of the 24-hour amplitude in airflow obstructions. Urinary-adrenaline and noradrenaline excretion after house dust mite (HDM) inhalation and after exercise was measured. Nine children with (group I), and nine without increased airflow obstruction overnight (group II) and nine age matched healthy children (group C) were included in the study. All patients showed an early obstructive reaction (EOR) after HDM challenge. Six children in group I and five in group II developed an EOR on exercise. A significant increase in urinary adrenaline excretion was observed after exercise in the control group (P < 0.05, values on the control and challenge day being 5.4 +/- 0.9 and 10.0 +/- 1.6 mumol/mol creat.). The same occurred for noradrenaline (P < 0.01, values being 28.2 +/- 2.5 and 49.0 +/- 5.7 mumol/mol creat.). Adrenergic response after both stimuli was impaired in the asthmatic groups, in group I more pronounced than in group II. Values from group I for adrenaline on the control day, HDM and exercise challenge were 6.0 +/- 0.8, 4.7 +/- 0.6, 6.0 +/- 1.0 and for noradrenaline 36.1 +/- 2.7, 27.2 +/- 2.3, 38.4 +/- 4.9 mumol/mol creat., respectively. Values from group II for adrenaline on these days were 5.6 42- 1.0, 3.7 42- 0.6 and 9.0 +/- 1.3 and for noradrenaline 28.3 +/- 3.2, 22.4 +/- 2.5, 41.3 +/- 5.9 mumol/mol creat., respectively.(ABSTRACT TRUNCATED AT 250 WORDS)