[Chemical neuro-anatomy of the thalamus: an example of the co-expression of 2 neuropeptides, cholecystokinin and intestinal vasoactive peptide]. / La neuroanatomie chimique du thalamus: exemple de coexpression de deux neuropeptides, cholécystokinine et peptide vasoactif intestinal.

Recent studies using hybridization histochemistry have demonstrated the presence of cholecystokinin (CCK) gene expression in thalamocortical and thalamo-striatal neurons. To further understand the chemical anatomy of the thalamus, we used the same method to study the coexpression of CCK and vasoactive intestinal peptide (VIP) genes in the same neurons. Most of the neurons expressing the VIP gene in the rat dorsal thalamus (found especially in the ventrolateral nucleus) also expressed the CCK gene, as demonstrated by autoradiography on emulsion-coated adjacent sections studied with probes recognizing either mRNA. Some further neurons, located in the reticular nucleus, had VIP mRNA at lower levels of expression but did not appear to contain CCK mRNA.

Distribution of neuropeptide Y-like immunoreactive fibers in the cat thalamus.

Neuropeptide Y-like immunoreactivity was studied in the thalamus of the cat using an indirect immunoperoxidase method. The densest network of immunoreactive fibers was observed in the nucleus (n.) paraventricularis anterior. In the anterior, intralaminar and midline thalamic nuclei, as well as in the n. geniculatum medialis, n. geniculatum lateralis, n. habenularis lateralis, n. medialis dorsalis, n. lateralis posterior and n. pulvinar a low density of neuropeptide Y-like immunoreactive fibers was observed. Neuropeptide Y-like fibers were totally absent in the n. ventralis lateralis, n. ventralis medialis, n. ventralis postero-medialis and n. ventralis postero-lateralis. In addition, neuropeptide Y-like perikarya were found in the n. parafascicularis, n. suprageniculatus, n. geniculatum lateralis ventralis, n. medialis dorsalis and n. lateralis posterior.

Effect of long-lasting diabetes mellitus on rat and human brain monoamines.

Experimental alloxan- or streptozotocin-produced diabetes in rats was accompanied by an increase in the levels of norepinephrine, dopamine, and serotonin, whereas the contents of metabolites, i.e., 5-hydroxyindoleacetic acid and homovanillic acid, in the whole brain gradually decreased with the duration of diabetes. Among the striatum, thalamus, and hypothalamus of alloxan diabetic rats, monoamine alterations were observed only in the hypothalamus; after 1 week an increase of norepinephrine content and after 13 weeks an increase of norepinephrine and dopamine contents were found. Tissues of 11 brain regions of 10 diabetic and 12 control patients post mortem were investigated for monoamine concentrations. Patients were all male, of similar age and interval between death and autopsy. Diabetic patients had an increase in the content of serotonin in the medial and lateral hypothalamus. The content of dopamine increased in the medial hypothalamus, putamen, and medial and lateral pallidus. In diabetic patients, the content of norepinephrine increased in the lateral pallidus and decreased in the nucleus accumbens and claustrum. Thus, it seems that diabetes mellitus in rats, as well as in humans is associated with regionally specific changes in brain monoamines.

Distribution of CCK mRNA in particular regions (hippocampus, periaqueductal grey and thalamus) of the rat by in situ hybridization.

Cholecystokinin (CCK) mRNA was detected by in situ hybridization at high magnification in some rat brain regions where CCK octapeptide (CCK-8) is thought to produce its pharmacological effects. The labeling of the dentate gyrus and the sparse but intensively stained cells found in the CA1 layer, stratum radiatum and hilus could correspond to interneurons involved in hippocampal neural activity, in agreement with excitatory responses induced by local injection of CCK-8. The intense labeling of the Edinger-Westphal nucleus and more generally the presence of CCK mRNA in the periaqueductal gray and thalamus ventrobasal nuclei could account for the various effects of CCK in pain transmission.

Distribution of serotonin-immunoreactive neurons in the brain of sockeye salmon fry.

The organization of the serotonergic cell groups in the brainstem of fishes and amphibians has received relatively little attention. It has been generally assumed that they are little differentiated and constitute a median cell column throughout the brainstem, and that laterally migrated serotonergic cell groups are largely lacking. In the present study we present evidence to the contrary. By the use of a sensitive immunocytochemical technique for the visualization of serotonin-immunoreactive (5HTir) neurons, we have been able to make a detailed delineation of the putatively serotonergic neuronal groups throughout the brain. In the epithalamus, 5HTir neurons were located in the left habenular nucleus in its dorsal subdivision. 5HTir neural elements, primarily photoreceptor cells, were present throughout the pineal organ and in some cases also in the parapineal organ. In the periventricular zones of the hypothalamus and posterior tuberculum, 5THir cerebrospinal fluid-contacting neurons were located in the paraventricular organ and in the dorsal, ventral and caudal zones of the periventricular hypothalamus. In the dorsal thalamus/synencephalon, 5THir neurons surround the tractus habenulo-interpeduncularis (fasciculus retroflexus). In the brainstem, several groups of 5HTir neurons could be discerned, that for reasons of topological similarity were named according to Lidov and Molliver a raphe pallidus/obscurus-complex (B1 and B2), raphe magnus (part of B3), median raphe (B8) possibly including raphe pontis (B5), raphe dorsalis (B4, B6 and B7), and B9. 5HTir neurons were observed in the central gray of the IVth ventricle, dorsal to the noradrenergic isthmal neurons and lateral to the brachium conjunctivum, in an area topologically equivalent with the dorsal subdivision of the locus coeruleus in mammals. In addition, small numbers of 5HTir neurons were located in the lobi faciales. Thus, the presence of well-differentiated groups of migrated serotonergic neurons is not an advanced trait of amniote brains, but may be a pattern common to all vertebrates.

[Contents of serotonin and 5-hydroxyindoleacetic acid in the rabbit brain after long-term administration of lithium oxybutyrate]. / Soderzhanie serotonina i 5-oksiindoluksusnoi kisloty v mozge krolikovpri dlitel'nom naznachenii oksibutirata litiia.

Serotonin (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA) levels in rat brain were analysed 24 hours after 7-, 15-, 29- days lithium hydroxybutyrate (LH) injections (10 mg/kg daily). After 7 days the drug reduced 5-HT in hypothalamus and 5-HIAA in the mid brain by 35%. After 15 days LH decreased 5-HT in striatum, hypothalamus by 32 and 17% and 5-HIAA in thalamus, hypothalamus by 28 and 44% respectively. After 29 days LH diminished 5-HT in striatum, hippocampus, amygdala by 24, 29 and 32% and 5-HIAA--in hypothalamus by 42%. The role of adaptative changes and stabilization processes in the central serotoninergic system in mechanism of LH psychotropic effects is discussed.

Neurokinin B and substance P genes are co-expressed in a subset of neurons in the rat habenula.

Colocalization of neurokinin B (NKB) and substance P (SP) mRNAs in neurons of the habenula was examined on thin, adjacent sections in the rat. Extensive colocalization was found in the medial habenula. In its dorsolateral part, most of the neurons contained both transcripts, with high levels of SP, while in the dorsomedial part fewer instances of colocalization were found. The ventral half could be divided into two parts, a dorsal part with most of the neurons containing both messages, having low levels of SP mRNA, and a ventral part with most of the cells containing only NKB mRNA. Cells in the medial habenula had low levels of NKB mRNA. These results suggest a structural and functional heterogeneity of the medial habenula.

[Prolactin-secreting neurons in the dorsolateral hypothalamus in Sprague-Dawley rats]. / Neurones à prolactine dans l'hypothalamus dorso-latéral du rat Sprague-Dawley.

By means of immunocytochemical techniques ovine prolactin like immunoreactivity (oPRL-LIR) has been demonstrated in the perikarya located around fornix in the dorso-lateral part of the rat hypothalamus. No PRL-LIR was observed in the arcuate n. perikarya. Immunoreactive fibers were present in the hypothalamus, medial thalamus, accumbens and amygdaloid nuclei.

[Distribution of the enkephalin neuron system in the rat thalamus; an immunohistochemical analysis].

Met-and Leu-enkephalin immunoreactive structures in the rat thalamus were investigated by indirect immunofluorescent method. Both structures showed similar distributions. Distribution: A large number of enkephalin-like immunoreactive (ENKI) terminals were observed in the antero-ventral nucleus (AV), the region between the parataenial nucleus, the antero-medial thalamic nucleus and ventral part of the ventral nucleus. ENKI cells were seen in the medial thalamic nucleus and ventral nucleus of the lateral geniculate body. Furthermore, ENKI cells are seen at the pars parvicellular of the posterior commissure and the lateral posterior thalamic nucleus detected only following pretreatment with colchicine. Fiber connection: The origin of ENKI terminals in the AV was examined by means of experimental manipulations. Injections of HRP (horseradish peroxidase) into the AV resulted in labeling of the MM (medial mammillary nucleus pars medialis) and MMC (medial mammillary nucleus pars centralis) but not the PM (premammillary nucleus). Destruction of the MM, where a number of ENKI neurons were seen, resulted in a disappearance of ENKI terminals in the ipsilateral AV. These findings suggest that MM is a major source of ENKI terminals in the AV.

Age-related changes of catecholamines and their metabolites content in the visual system of the rat.

The changes in the content of the catecholamines in each structure of the geniculate and extrageniculate visual system of the rat during the aging period (6-30 months) have been studied. Dopamine was found at lower levels than noradrenaline in all the structures. The dopamine and noradrenaline showed different developmental profiles. Dopamine and its metabolite levels decreased in the lateral geniculate and visual cortex and increased in superior colliculus and posterior thalamus. Noradrenaline and its metabolites increased in all structures during the aging period. However, 3-methoxy-4-hydroxyphenylglycol/noradrenaline and normetanephrine/noradrenaline ratios decreased in all structures except in superior colliculus. These results suggest age-related changes in the catecholamines in the visual system of the rat.

Characterization and localization of a novel neuroreceptor for the peptide sarafotoxin.

We have recently shown that the rat atrium and brain contain specific high affinity receptors for the novel snake vasoconstrictor peptide sarafotoxin-b (SRTXb), and demonstrated toxin-induced phosphoinositide hydrolysis. Here we report on the characteristics of 125I-SRTXb receptors and their regional distribution in rat brain. 125I-SRTX receptors in the rat brain bind the toxin rapidly and with high affinity. The binding was not inhibited by ligands of known neurotransmitter receptor and ion channels. 125I-SRTX receptors have a distinctive regional distribution. The highest densities were observed in the cerebellum, thalamus and hypothalamus (850, 550 and 450 fmol/mg protein, respectively) and the lowest densities in the caudate and cerebral cortex (82 and 62 fmol/mg protein, respectively). Taken together our results suggest that mammalian brains contain a hitherto undetected neuroreceptor that may operate in neurotransmission with a "SRTX-like" brain peptide, similar to the SRTX homologous vasoconstrictor peptide of the mammalian endothelium endothelin.

Regional distribution of the GABAA/benzodiazepine receptor (alpha subunit) mRNA in rat brain.

A human cDNA clone containing the 5' coding region of the GABAA/benzodiazepine receptor alpha subunit was used to quantify and visualize receptor mRNA in various regions of the rat brain. Using a [32P]CTP-labelled antisense RNA probe (860 bases) prepared from the alpha subunit cDNA, multiple mRNA species were detected in Northern blots using total and poly A rat brain RNA. In all brain regions, mRNAs of 4.4 and 4.8 kb were observed, and an additional mRNA of 3.0 kb was detected in the cerebellum and hippocampus. The level of GABAA/benzodiazepine receptor mRNA was highest in the cerebellum followed by the thalamus = frontal cortex = hippocampus = parietal cortex = hypothalamus much greater than pons = striatum = medulla. In situ hybridization revealed high levels of alpha subunit mRNA in cerebellar gray matter, olfactory bulb, thalamus, hippocampus/dentate gyrus, and the arcuate nucleus of the hypothalamus. These data suggest the presence of multiple GABAA/benzodiazepine receptor alpha subunit mRNAs in rat brain and demonstrate the feasibility of studying the expression of genes encoding the GABAA/benzodiazepine receptor after pharmacological and/or environmental manipulation.

Identification and quantification of kynurenic acid in human brain tissue.

Serial ion-exchange and high-performance liquid chromatography separations were employed for the tissue extraction and purification of kynurenic acid (KYNA). Subsequently, the compound isolated from postmortem human brain tissue was unequivocally identified as KYNA by nuclear magnetic resonance and mass spectrometric analyses. Regional distribution analyses revealed the highest concentration of KYNA (1.58 +/- 0.43 pmol/mg tissue) in the caudate nucleus with lower levels in the thalamus, globus pallidus, hippocampus, parietal cortex and frontal cortex. Of the brain structures examined, the lowest concentration of KYNA (0.14 +/- 0.02 pmol/mg tissue) was found in the cerebellum.

The effects of N-acetylaspartylglutamate and distribution of N-acetylaspartylglutamate-like immunoreactivity in the rat somatosensory thalamus.

The ventrobasal thalamus and adjacent regions were stained for the presence of N-acetylaspartylglutamate-like immunoreactivity. Immunoreactive axonal terminals were observed in this area and also in certain non-specific thalamic nuclei, the reticular thalamic nucleus and the lateral geniculate nucleus. Stained somata were found in the habenula, centrolateral thalamic nucleus and reticular thalamic nucleus. Iontophoretically applied N-acetylaspartylglutamate had variable, although predominantly inhibitory, actions on ventrobasal thalamus neurons. These results indicate that N-acetylaspartylglutamate is unlikely to be the neurotransmitter of ascending somatosensory afferents, but do not rule out the possibility that it has some other neurotransmitter or neuromodulator role in the ventrobasal thalamus.

Distribution of cholecystokinin-immunoreactive cell bodies in the male and female rat: II. Bed nucleus of the stria terminalis and amygdala.

The distribution of cholecystokinin-immunoreactive (CCK-I) cell bodies was studied in the bed nucleus of the stria terminalis (BST) and amygdaloid complex of colchicine-treated male and female rats. Immunoreactive cells were visualized in the BST medial amygdaloid (MeA), central lateral, basolateral, basolateral ventral, medial, intercalated, anterior cortical, and posterior cortical nuclei and the amygdalohippocampal zone. Several significant sex differences were observed. In the male, a dense aggregation of CCK-I cell bodies was visualized in the MeA, especially in the dorsocaudal part and in the encapsulated part of the BST. In comparison, female rats had relatively fewer immunoreactive cells in both of these regions. In the lateral and basolateral amygdaloid nuclei, however, more CCK-I cells were visualized in the female than in the male, but the difference was not statistically significant. These data provide characterization of a sexually differentiated CCK system. In addition, we observed that the number of CCK-I cells in the BST and posterodorsal part of the MeA was substantially reduced after castration. The number of CCK-I cells in female rats, however, was not significantly reduced after ovariectomy in any of the regions studied. These findings imply that the steroid regulation of CCK is sexually differentiated. The sexually dimorphic distribution of CCK-I cells in areas that are targets of steroid hormones and regulate reproductive processes is consistent with the possibility that CCK participates in central integration of sensory and steroidal input that modulates reproductive behavior.

[Biochemical and immunocytochemical study of the axonal domain of mature and immature microtubules]. / Etude biochimique et immunocytochimique du domaine axonal des microtubules matures et immatures.

The axonal domain of microtubules was analyzed using biochemistry and immunocytochemistry. High resolution isofocusing was used to study the cold-labile and cold-stable fractions of various samples: thalamus, optic nerve, sciatic nerve, brachial plexus and trigeminal nerve. The cold-labile fraction from the thalamus, i.e. from a central nervous system cell population contains 20 well-resolved isotypes as the axonal fraction from the central nervous system. The cold-stable fraction from the peripheral nervous system contains only 18 isotypes. All the cold-stable fractions are characterized by an important relative quantity of isotypes 7-9, with specific patterns tissue dependent. The cold-labile fractions are specific in both the central and peripheral nervous system. Immunocytochemistry using anti-tubulin and anti-MAP2 specific antibodies was used in PEG semi-thin sections of the embryonic tectal plate at stage E10. In the axonal profiles, cold-stable fragments of microtubules were observed. The perikaryon of the young neurons, both migrating and post-migratory, contains MAP2 as the cold-labile fractions of microtubules of the axonal process. These results suggest that; tubulin is involved in the cold-stability of the axonal domain, but this role seems tissue dependent; MAP2 is a marker of neuronal differentiation; in the growing axonal process, cold-stable fragments are present and the cold-labile pool of microtubules contain a transiently expressed protein, MAP2.

Identification and localization of glucagon-like peptide-1 and its receptor in rat brain.

The existence and distribution of glucagon-like peptide-1 (GLP-1) and its receptor in rat brain in relation to that of glucagon were examined. The concentration of GLP-1 immunoreactivity (GLP-1-IR), measured by a specific and sensitive RIA established in this study with anti GLP-1 serum (LMT-01), was found to be highest in the thalamus-hypothalamus, followed by the medulla oblongata. The distribution of glucagon-like immunoreactivity was similar to that of GLP-1-IR. However, appreciable glucagon immunoreactivity was detected only in the thalamus-hypothalamus. Gel filtration analysis showed the presence of GLP-1-IR of various molecular weights in the extract of thalamus-hypothalamus including that eluted at the same position as synthetic GLP-1 (1-37); moreover, HPLC analysis also confirmed the presence of GLP-1-IR, eluted at the exact position as synthetic GLP-1 (1-37). The distribution of receptors for GLP-1 corresponded with that of GLP-1-IR in the rat brain, except in the pituitary gland. The distribution of these receptors was also similar to that of glucagon receptors. The thalamus-hypothalamus, pituitary gland, and medulla oblongata were rich in GLP-1 and glucagon-binding sites. The binding affinities of GLP-1 and glucagon were in the nanomolar range [disocciation constant Kd approximately equal to 4 nM]. The presence of specific, high affinity receptors for GLP-1 was confirmed by demonstrating that GLP-1 stimulated cAMP formation in the thalamus-hypothalamus and the pituitary gland. The concentration of GLP-1 required for half-maximal stimulation of cAMP formation in these regions was about 1 nM. These results suggest that GLP-1 may be synthesized in certain parts of the brain and play a role as a neurosignal transmitter.

Distribution of neurotensin-like immunoreactivity in the diencephalon of the Japanese monkey (Macaca fuscata).

The distribution of neurotensin-like immunoreactive (NT-LI) neurons was examined in the thalamus and hypothalamus of the Japanese monkey (Macaca fuscata) by using the peroxidase-antiperoxidase immunocytochemistry technique. In the thalamus, NT-LI neuronal perikarya were distributed mainly in the midline nuclear group and the dorsomedial nucleus, and partially in the intralaminar nucleus: Immunoreactive fibers were mainly distributed in the midline nucleus, particularly in the nucleus rhomboidalis. Numerous immunoreactive fibers were also detected in the regions that contain the pathways to extrathalamic areas such as the stratum zonale and inferior thalamic peduncle. In the hypothalamus, many immunoreactive neuronal perikarya were distributed in the lateral hypothalamic area and in the arcuate nucleus. Immunoreactive fibers were disseminated throughout the hypothalamus, but they were dense in the preoptic area and sparse in the ventromedial nucleus. An accumulation of dense immunoreactive endings was also observed in the external layer of the median eminence. NT-LI fibers in the external layer of the median eminence were considered to represent nerve endings near portal vessels. Functional roles of neurotensin in the thalamus and hypothalamus are discussed from the anatomical point of view.

Local circuit neurons in the rat ventrobasal thalamus--a GABA immunocytochemical study.

The ventrobasal thalamus of seven rats was processed for immunocytochemistry using antisera to glutamate decarboxylase or gamma-aminobutyrate (GABA). Glutamate decarboxylase-stained sections showed a network of stained fibers and terminals but no stained cell bodies. GABA-stained sections had fewer stained fibers and terminals but did show a few stained cell bodies. Cell bodies were especially apparent when carbazole was used for a chromogen for the peroxidase-antiperoxidase visualization. The GABA-stained cells were found to be distributed throughout the ventrobasal complex, to have smaller soma cross-sectional areas than most other cells (81 +/- 34 microns vs 105 +/- 36 microns for all cells) and to make up 0.4 +/- 0.3% of the neuronal population of the ventrobasal complex. Injections of horseradish peroxidase into the somatosensory cortex (SI) retrogradely filled many neurons in the ventrobasal thalamus, but none of these labeled neurons were double labeled with GABA. These results indicate that the GABA-labeled cells probably represent a small population of local circuit neurons in the rat ventrobasal thalamus.