Chondroitin sulfate proteoglycans in the rat thalamus: expression during postnatal development and correlation with calcium-binding proteins in adults

Postnatal expression of chondroitin sulfate proteoglycans was studied in the rat thalamus by immunocytochemistry and Western immunoblotting techniques with monoclonal antibodies that recognize carbohydrate epitopes (clones CS-56, 1-B-5, 2-B-6). The complex of the results shows that these antibodies recognize mostly nonoverlapping molecules whose expression is regulated during postnatal development. Chondroitin sulfate proteoglycans, recognized by antibody CS-56, and hyaluronan, identified by antibody 1-B-5 after hyaluronidase digestion, are abundant in the neuropil of most thalamic nuclei at the perinatal stage and progressively decrease during the second week of life, attaining levels barely detectable by immunocytochemistry at the end of the third week. In adult thalamus, chondroitin sulfate proteoglycans of high molecular mass, bearing glycosaminoglycans unsulfated in the linking region, and recognized by antibody 1-B-5 are confined to perineuronal nets around neurons chiefly localized in thalamic reticular nucleus. The immunoreactvity for antibody 2-B-6, specific for chondroitin-4-sulfate, is low at the perinatal stage and is not detectable in adult thalamus. Double-immunolabeling has shown that, along the rostrocaudal extension of reticular nucleus, the most developed perineuronal nets are associated with a subset of neurons expressing calretinin, and not with parvalbumin-positive neurons, which represent the largest neuronal population of the nucleus. The distribution of perineuronal nets supports the presence, in thalamic reticular nucleus, of neuronal subpopulations with different morphological and physiological features.     

Development, neurochemical properties, and axonal projections of a population of last-order premotor interneurons in the white matter of the chick lumbosacral spinal cord

There is general agreement that last-order premotor interneurons-a set of neurons that integrate activities generated by the spinal motor apparatus, sensory information and volleys arising from higher motor centres, and transmit the integrated signals to motoneurons through monosynaptic contacts-play crucial roles in the initiation and maintenance of spinal motor activities. Here, we demonstrate the development, neurochemical properties, and axonal projections of a unique group of last-order premotor interneurons within the ventrolateral aspect of the lateral funiculus of the chick lumbosacral spinal cord. Neurons expressing immunoreactivity for neuron-specific enolase were first detected in the ventrolateral white matter at embryonic day 9 (E9). The numbers of immunoreactive neurons were significantly increased at E10-E12, while most of them were gradually concentrated in small segmentally arranged nuclei (referred to as major nuclei of Hofmann) protruding from the white matter in a necklace like fashion dorsal to the ventral roots. The major nuclei of Hofmann became more prominent at E12-E16, but substantial numbers of cells were still located within the ventrolateral white matter (referred to as minor nucleus of Hofmann). The distribution of immunoreactive neurons achieved by E16 was maintained during later developmental stages and was also characteristic of adult animals. After injection of Phaseolus vulgaris-leucoagglutinin unilaterally into the minor nucleus of Hofmann, labeled fibres were detected in the ventrolateral white matter ipsilateral to the injection site. Ascending and descending fibres were revealed throughout the entire rostro-caudal length of the lumbosacral spinal cord. Axon terminals were predominantly found within the lateral motor column and the ventral regions of lamina VII ipsilateral to the injection site. Several axon varicosities made close appositions with somata and dendrites of motoneurons, which were identified as synaptic contacts in a consecutive electron microscopic study. With the postembedding immunogold method, 21 of 97 labeled terminals investigated were immunoreactive for glycine and 2 of them showed immunoreactivity for gamma-aminobutyric acid (GABA). The axon trajectories of neurons within the minor nucleus of Hofmann suggest that some of these cells might represent a population of last-order premotor interneurons. J. Exp. Zool. 286:157-172, 2000.     

Differential expression of several molecules of the extracellular matrix in functionally and developmentally distinct regions of rat spinal cord

We have examined the regional distribution of several chondroitin sulfate proteoglycans (neurocan, brevican, versican, aggrecan, phosphacan), of their glycosaminoglycan moieties, and of tenascin-R in the spinal cord of adult rat. The relationships of these molecules with glial and neuronal populations, identified with appropriate markers, were investigated by using multiple fluorescence labeling combined with confocal microscopy. The results showed that the distribution of the examined molecules was similar at all spinal cord levels but displayed area-specific differences along the dorso-ventral axis, delimiting functionally and developmentally distinct areas. In the gray matter, laminae I and II lacked perineuronal nets (PNNs) of extracellular matrix and contained low levels of chondroitin sulfate glycosaminoglycans (CS-GAGs), brevican, and tenascin-R, possibly favoring the maintenance of local neuroplastic properties. Conversely, CS-GAGs, brevican, and phosphacan were abundant, with numerous thick PNNs, in laminae III-VIII and X. Motor neurons (lamina IX) were surrounded by PNNs that contained all molecules investigated but displayed various amounts of CS-GAGs. Double-labeling experiments showed that the presence of PNNs could not be unequivocally related to specific classes of neurons, such as motor neurons or interneurons identified by their expression of calcium-binding proteins (parvalbumin, calbindin, calretinin). However, a good correlation was found between PNNs rich in CS-GAGs and the neuronal expression of the Kv3.1b subunit of the potassium channel, a marker of fast-firing neurons. This observation confirms the correlation between the electrophysiological properties of these neurons and the specific composition of their microenvironment.     

Distribution of dopamine immunoreactive systems in brain stem and spinal cord of the chameleon

An immunohistochemical method, using glutaraldehyde fixation and a highly specific monoclonal antibody recently synthetized against dopamine (DA)-glutaraldehyde protein conjugate, permitted direct visualization of DA structures in the brainstem and spinal cord of a reptile (Chameleon). DA-immunoreactive cell bodies occurred in some contiguous areas of the midbrain tegmentum. The first one was located in the ventral tegmental area. Some somata intermingled with the oculomotor nucleus. The second group was the large round or oval DA-Immunostained neurons located in the substantia nigra. More caudally, a third group of round or fusiform DA-cell bodies was seen in an homologous area of so called mammalian A8 and were continuous with the substantia nigra group. In the medulla oblongata, the DA-containing cells were shown in the nucleus of solitary tract and in the dorsal lateral part of the dorsal motor nucleus of the vagus. The density of this DA-Immunoreactive neurons decreased more caudally. At the medullo-spinal level and upper cervical spinal cord, a few labelled cells were distinguished near the central canal. In the spinal cord DA-immunopositive cell bodies were observed in the vicinity of the central canal and formed a continuous column that extended throughout the rostral spinal cord. The apical processes of these neurons seemed to be in contact with the lumen of the central canal. This study constitute the first visualization of the immunoreactive DA-cell bodies at the medullo-spinal level which were already described, as TH immunoreactive in other species of reptiles.     

SPARCL1-containing neurons in the human brainstem and sensory ganglion

Secreted protein, acidic and rich in cysteine-like 1 (SPARCL1) is a member of the osteonectin family of proteins. In this study, immunohistochemistry for SPARCL1 was performed to obtain its distribution in the human brainstem, cervical spinal cord, and sensory ganglion. SPARCL1-immunoreactivity was detected in neuronal cell bodies including perikarya and proximal dendrites, and the neuropil. The motor nuclei of the IIIrd, Vth, VIth, VIIth, IXth, Xth, XIth, and XIIth cranial nerves and spinal nerves contained many SPARCL1-immunoreactive (-IR) neurons with medium-sized to large cell bodies. Small and medium-sized SPARCL1-IR neurons were distributed in sensory nuclei of the Vth, VIIth, VIIIth, IXth, and Xth cranial nerves. In the medulla oblongata, the dorsal column nuclei also had small to medium-sized SPARCL1-IR neurons. In addition, SPARCL1-IR neurons were detected in the nucleus of the trapezoid body and pontine nucleus within the pons and the arcuate nucleus in the medulla oblongata. In the cervical spinal cord, the ventral horn contained some SPARCL1-IR neurons with large cell bodies. These findings suggest that SPARCL1-containing neurons function to relay and regulate motor and sensory signals in the human brainstem. In the dorsal root (DRG) and trigeminal ganglia (TG), primary sensory neurons contained SPARCL1-immunoreactivity. The proportion of SPARCL1-IR neurons in the TG (mean?±?SD, 39.9?±?2.4%) was higher than in the DRG (30.6?±?2.1%). SPARCL1-IR neurons were mostly medium-sized to large (mean?±?SD, 1494.5?±?708.3?μm²; range, 320.4–4353.4?μm²) in the DRG, whereas such neurons were of various cell body sizes in the TG (mean?±?SD, 1291.2?±?532.8?μm²; range, 209.3–4326.4?μm²). There appears to be a SPARCL1-containing sensory pathway in the ganglion and brainstem of the spinal and trigeminal nervous systems.     

Brain natriuretic peptide (BNP)-like immunoreactivity in the central nervous system of the crested newt.

The distribution of brain natriuretic peptide (BNP)-like immunoreactivity (ir) was studied in the brain of a urodele amphibian, the crested newt Triturus carnifex Laur. BNP-like immunoreactive neurons were found mainly in the caudal hypothalamus (retro- and supra-chiasmatic areas) and in the preoptic area. A widespread innervation throughout the brainstem as far as the spinal cord was also observed. By double immunostaining (after section incubation with a-BNP and a-tyrosine hydroxylase-TH-antibodies), close topographical relationships between BNP-like and TH-like immunoreactive neurons within the hypothalamus were found.     

New data on the immunocytochemical localization of thyrotropin-releasing hormone in the rat central nervous system

An antiserum raised against the synthetic tripeptide pyroglutamyl-histidyl-proline (free acid) was used to localize thyrotropin-releasing hormone (TRH) in the rat central nervous system (CNS) by immunocytochemistry. The distribution of TRH-immunoreactive structures was similar to that reported earlier; i.e., most of the TRH-containing perikarya were located in the parvicellular part of the hypothalamic paraventricular nucleus, the suprachiasmatic portion of the preoptic nucleus, the dorsomedial nucleus, the lateral basal hypothalamus, and the raphe nuclei. Several new locations for TRH-immunoreactive neurons were also observed, including the glomerular layer of the olfactory bulb, the anterior olfactory nuclei, the diagonal band of Broca, the septal nuclei, the sexually dimorphic nucleus of the preoptic area, the reticular thalamic nucleus, the lateral reticular nucleus of the medulla oblongata, and the central gray matter of the mesencephalon. Immunoreactive fibers were seen in the median eminence, the organum vasculosum of the lamina terminalis, the lateral septal nucleus, the medial habenula, the dorsal and ventral parabrachial nuclei, the nucleus of the solitary tract, around the motor nuclei of the cranial nerves, the dorsal vagal complex, and in the reticular formation of the brainstem. In the spinal cord, no immunoreactive perikarya were observed. Immunoreactive processes were present in the lateral funiculus of the white matter and in laminae V-X in the gray matter. Dense terminal-like structures were seen around spinal motor neurons. The distribution of TRH-immunoreactive structures in the CNS suggests that TRH functions both as a neuroendocrine regulator in the hypothalamus and as a neurotransmitter or neuromodulator throughout the CNS.     

Response of NADPH-Diaphorase-Exhibiting Neurons in the Medullar Reticular Formation to High Spinal Cord Injury

1. The effect of hemisection of the cervical spinal cord on NADPH-diaphorase staining in the reticular nuclei of the rabbit medulla was investigated using histochemical technique.2. A quantitative assessment of somal and neuropil NADPH-diaphorase staining was made by an image analyzer in a selected area of each reticular nucleus of the rabbit medulla.3. On the 7th postsurgery day, the highest up-regulation of somatic NADPH-diapho- rase staining was observed in regions regulating cardiorespiratory processes; however, the highest increase of neuropil NADPH-diaphorase staining was found in the reticular nuclei modulating the tonus of postural muscles.4. The degeneration of non-NADPH-diaphorase-stained neurons was detected throughout the reticular formation of the medulla, but the extent of neuronal death did not correlate with the up-regulation of the NADPH-diaphorase staining in the reticular nuclei of the medulla.5. The findings provide evidence that NADPH-diaphorase-exhibiting neurons are refractory to the hemisection of the cervical spinal cord and that the neuronal up-regulation of NADPH-diaphorase at the medullar level is probably not a causative factor leading to the death of the reticulospinal neurons.     

Oxidative Stress and Autophagic Alteration in Brainstem of SOD1-G93A Mouse Model of ALS

Amyotrophic lateral sclerosis (ALS) is a progressive and fatal neurodegenerative disease involving both upper and lower motor neurons. The mechanism of motor neuron degeneration is still unknown. Although many studies have been performed on spinal motor neurons, few have been reported on brainstem and its motor nuclei. The aim of this study was to investigate oxidative stress and autophagic changes in the brainstem and representative motor nuclei of superoxide dismutase 1 (SOD1)-G93A mouse model of ALS. The expression levels of cluster of differentiation molecule 11b (CD11b), glial fibrillary acidic protein, glutamate–cysteine ligase catalytic subunit, heme oxygenase-1, NAD(P)H: quinone oxidoreductase 1, voltage-dependent anion-selective channel protein 1, Sequestosome 1/p62 (p62), microtubule-associated protein 1 light chain 3B (LC3), and SOD1 proteins in brainstem were examined by Western blot analysis. Immunohistochemistry and immunofluorescence were performed to identify the cellular localization of SOD1, p62, and LC3B, respectively. The results showed that there were progressive asctrocytic proliferation and microglial activation, induction of antioxidant proteins, and increased p62 and LC3II expression in brainstem of SOD1-G93A mice. Additionally, SOD1 and p62 accumulated in hypoglossal, facial, and red nuclei, but not in oculomotor nucleus. Furthermore, electron microscope showed increased autophagic vacuoles in affected brainstem motor nuclei. Our results indicate that brainstem share similar gliosis, oxidative stress, and autophagic changes as the spinal cord in SOD1-G93A mice. Thus, SOD1 accumulation in astrocytes and neurons, oxidative stress, and altered autophagy are involved in motor neuron degeneration in the brainstem, similar to the motor neurons in spinal cord. Therefore, therapeutic trials in the SOD1G93A mice need to target the brainstem in addition to the spinal cord.     

大鼠延髓网状背侧亚核内5-羟色胺能、P物质样和脑啡肽样阳性终末的中枢起源

研究用荧光金（ＦＧ）逆行追踪与免疫荧光组化染色相结合的双标技术对大鼠脑干向延髓网状背侧亚核（ＳＲＤ）的５┐羟色胺（５┐ＨＴ）能、Ｐ物质（ＳＰ）能和亮氨酸┐脑啡肽（Ｌ┐ＥＮＫ）能投射进行了观察。将ＦＧ注入ＳＲＤ后,ＦＧ逆标神经元主要见于中脑导水管周围灰质、脑干中缝核簇（中缝背核、中缝正中核、中缝桥核、中缝大核、中缝隐核和中缝苍白核）、巨细胞网状核α部、延髓网状结构的内侧部和外侧部、延髓外侧网状核、三叉神经脊束核尾侧亚核和孤束核。５┐羟色胺（５┐ＨＴ）样、Ｐ物质（ＳＰ）样和亮氨酸脑啡肽（Ｌ┐ＥＮＫ）样阳性神经元主要见于中脑导水管周围灰质、脑干中缝核簇和巨细胞网状核α部;此外,ＳＰ样和Ｌ┐ＥＮＫ样阳性神经元还见于臂旁核、背外侧被盖核和孤束核。ＦＧ逆标并呈５┐ＨＴ样、ＳＰ样或Ｌ┐ＥＮＫ样阳性的双标神经元也主要见于中脑导水管周围灰质、脑干中缝核簇和巨细胞网状核α部,尤其是位于延髓中缝核团内的双标神经元数量较多。本研究的结果说明ＳＲＤ内的５┐ＨＴ样、ＳＰ样和Ｌ┐ＥＮＫ样阳性终末主要来自中脑导水管周围灰质、脑干中缝核簇和巨细胞网状核α部,向ＳＲＤ发出５┐ＨＴ能、ＳＰ能和Ｌ┐ＥＮＫ能投射的上述核团对ＳＲＤ发挥“弥漫性伤害抑     

Immunocytochemical Localization of TASK-3 Channels in Rat Motor Neurons

Motor neurons are large cholinergic neurons located in the brain stem and spinal cord. In recent years, a functional role for TASK channels in cellular excitability and vulnerability to anesthetics of motor neurons has been described. Using a polyclonal monospecific antibody against the tandem pore domain K⁺ channel (K2P channel) TWIK-related acid-sensitive K⁺ channel (TASK-3), we analyzed the expression of the TASK-3 protein in motor systems of the rat CNS. Immunocytochemical staining showed strong TASK-3 expression in motor neurons of the facial, trigeminal, ambiguus, and hypoglossal nuclei. Oculomotor nuclei (including trochlear and abducens nucleus) were also strongly positive for TASK-3. The parasympathetic Edinger-Westphal nucleus and dorsal vagal nucleus showed significant, but weaker expression compared with somato- and branchiomotoric neurons. In addition, motor neurons in the anterior horn of the spinal cord were also strongly labeled for TASK-3 immunoreactivity. Based on morphological criteria, TASK-3 was found in the somatodendritic compartment of motor neurons. Cellular staining using methyl green and immunofluorescence double-labeling with anti-vesicular acetylcholine transporter (anti-vAChT) indicated ubiquitous TASK-3 expression in motor neurons, whereas in other brain regions TASK-3 showed a widespread but not ubiquitous expression. In situ hybridization using a TASK-3 specific riboprobe verified the expression of TASK-3 in motor neurons at the mRNA level.     

Expression of major histocompatibility complex antigens and CR3 complement receptors in activated microglia following an injection of ricin into the sciatic nerve in rats.

The ventral horn motor neurons in the lower lumbar cord underwent rapid degeneration following an injection of Ricinus communis agglutinin-60 (RCA) into the sciatic nerve. The cell death which was most drastic between the fifth and seventh post-injection day elicited a significant increase in the number of microglia. The activated microglia were scattered throughout the neuropil but the dramatic feature was their close association with the somata of the degenerating neurons. Often several microglial cells were seen surrounding the soma of a degenerating neuron. Immunocytochemical study showed that both the interstitial as well as the perineuronal activated microglia were labelled with the monoclonal antibodies OX-18 and OX-42 for the detection of MHCI encoded antigen and type three complement receptors, respectively. Intense immunoreactivity was observed especially in the perineuronal microglia with OX-18. Electron microscopic study confirmed the identification of the activated microglia. Although the activated microglia closely apposed the neuronal soma, there was no sign of a direct endocytosis. The cytoplasm of the activated microglia, however, contained massive lipofuscin bodies in longer survival animals. Electron microscopic immunocytochemical study showed that the immunoreactivity of the activated microglia was localized along their plasma membrane facing the neuronal soma. Since the microglia cells on the contralateral side of the ventral horn were not marked by the antibodies used, it was postulated that the vigorous expression of MHCI antigen and CR3 receptors on the activated microglia was induced by the neuronal degeneration resulting from the application of the toxin ricin.     

Distribution of FMRFamide-like immunoreactivity in the central nervous system of the Formosan monkey (Macaca cyclopsis)

The distribution of FMRFamide-like immunoreactivity in the central nervous system of the Formosan monkey (Macaca cyclopsis) was investigated employing immunohistochemical techniques. FMRFamide-containing cells were found to be widely distributed throughout the forebrain. Principal densities of FMRFamide neuronal perikarya were observed in the following areas: the amygdaloid complex, the olfactory tubercle, the cerebral cortex, the basal ganglia, the septum, the caudate-putamen and the arcuate nucleus. A large number of immunoreactive fibers were observed in areas ranging from the cerebral cortex to the spinal cord, and were noted in the following locations: the preoptic area, the tuberal and posterior hypothalamic areas, the bed nucleus of the stria terminalis, the nuclei of the spinal trigeminal nerve, the hypoglossal nucleus, the nucleus of the solitary tract, and the dorsal horn of the spinal cord. The results generally parallel those described in the rat and guinea pig.     

The distribution of cholinergic neurons in the human central nervous system

Choline acetyltransferase (ChAT), the enzyme responsible for the biosynthesis of acetylcholine, is presently the most specific marker for identifying cholinergic neurons in the central and peripheral nervous systems. The present article reviews immunohistochemical and in situ hybridization studies on the distribution of neurons expressing ChAT in the human central nervous system. Neurons with both immunoreactivity and in situ hybridization signals of ChAT are observed in the basal forebrain (diagonal band of Broca and nucleus basalis of Meynert), striatum (caudate nucleus, putamen and nucleus accumbens), cerebral cortex, mesopontine tegmental nuclei (pedunculopontine tegmental nucleus, laterodorsal tegmental nucleus and parabigeminal nucleus), cranial motor nuclei and spinal motor neurons. The cerebral cortex displays regional and laminal differences in the distribution of neurons with ChAT. The medial septal nucleus and medial habenular nucleus contain immunoreactive neurons for ChAT, which are devoid of ChAT mRNA signals. This is probably because there is a small number of cholinergic neurons with a low level of ChAT gene expression in these nuclei of human. Possible connections and speculated functions of these neurons are briefly summarized.     

Cystathionine beta-synthase in the structural elements of the human brain and spinal cord

Investigated by immunohistochemistry the presence and distribution of CBS in the nuclei of the spinal cord and brainstem in 8 mens of 18-44 years old, died from mechanical injury is not related to the central nervous system damage. Established that the CBS-neurons are revealed in all parts of stem and spinal cord, but their content in the studied cores, ranging from 0.9 to 17 %. Observed a definite relationship between the activity of CBS and the quantity of neurons. Large cells of motor nuclei often have a high and very high intensity of the reaction. In the sensory nuclei of a high proportion of small neurons with low activity of the enzyme.     

Calcitonin gene-related peptide: detailed immunohistochemical distribution in the central nervous system

With the use of an antiserum generated in rabbits against synthetic human calcitonin gene-related peptide (CGRP) the distribution of CGRP-like immunoreactive cell bodies and nerve fibers was studied in the rat central nervous system. A detailed stereotaxic atlas of CGRP-like neurons was prepared. CGRP-like immunoreactivity was widely distributed in the rat central nervous system. CGRP positive cell bodies were observed in the preoptic area and hypothalamus (medial preoptic, periventricular, anterior hypothalamic nuclei, perifornical area, medial forebrain bundle), premamillary nucleus, amygdala medialis, hippocampus and dentate gyrus, central gray and the ventromedial nucleus of the thalamus. In the midbrain a large cluster of cells was contained in the peripeduncular area ventral to the medial geniculate body. In the hindbrain cholinergic motor nuclei (III, IV, V, VI, VII XII) contained CGRP-immunoreactivity. Cell bodies were also observed in the ventral tegmental nucleus, the parabrachial nuclei, superior olive and nucleus ambiguus. The ventral horn cells of the spinal cord, the trigeminal and dorsal root ganglia also contained CGRP-immunoreactivity. Dense accumulations of fibers were observed in the amydala centralis, caudal portion of the caudate putamen, sensory trigeminal area, substantia gelatinosa, dorsal horn of the spinal cord (laminae I and II). Other areas containing CGRP-immunoreactive fibers are the septal area, nucleus of the stria terminalis, preoptic and hypothalamic nuclei (e.g., medial preoptic, periventricular, dorsomedial, median eminence), medial forebrain bundle, central gray, medial geniculate body, peripeduncular area, interpeduncular nucleus, cochlear nucleus, parabrachial nuclei, superior olive, nucleus tractus solitarii, and in the confines of clusters of cell bodies. Some fibers were also noted in the anterior and posterior pituitary and the sensory ganglia. As with other newly described brain neuropeptides it can only be conjectured that CGRP has a neuroregulatory action on a variety of functions throughout the brain and spinal cord.     

Expression of Nerve Growth Factor,p75, and trk in the Somatosensory and Motor Cortices of Mature Rats: Evidence for Local Trophic Support Circuits

Neurotrophins such as nerve growth factor (NGF) are critical for the maintenance of CNS neurons. We determined the expression of NGF and the neurotrophin receptors p75 and trk in the somatosensory and motor cortices of mature rats with immuno-histochemical techniques. Sections of mature rat cortex were processed immunohisto-chemically with primary antibodies directed against NGF, p75, or trk. The distribution of immunoreactive elements was examined, and stereological techniques were used to determine the density and size of immunoreactive cell bodies. Some sections processed for trk immunoreactivity were examined with an electron microscope.

From the size and morphology of the labeled cells, it appeared that only neurons in the gray matter were NGF-positive. NGF was detected in one-third of the neurons in layers II-III, V, and VI of both somatosensory cortex and motor cortex; however, fewer than 1 in 12 of the layer IV neurons was NGF-positive. With the notable exception of layer V, few cell bodies (2–10% of the total population) were p75– or trk-immunoreactive. Layer Vb was replete with receptor-positive cell bodies; more than one-third of the layer Vb neurons were p75– or trk-positive. All labeled cells appeared to be pyramidal neurons. The distribution of p75 labeling with the two anti-p75 antibodies was indistinguishable. In addition, the neuropil in the supragranular laminae was p75– or trk-positive. Electron microscopy showed that trk immunoreactivity was also expressed by dendrites. Only rarely were immunoreactive axons detected.

In summary, NGF is expressed by cortical neurons throughout cortex, and neurotrophin receptors are widely produced by postsynaptic targets. Thus, NGF appears to participate in an intracortical autoregulatory system. The strong expression of neurotrophin receptors by pyramidal neurons in layer Vb (the origin of brainstem and spinal cord projections) suggests that the neurotrophins are especially critical for the regulation of corticofugal projection systems.     

A light and electron microscopic study of the GABA transporter GAT-3 in the monkey basal ganglia and brainstem

The present study aimed to elucidate the distribution of the GABA transporter GAT-3 in the monkey basal ganglia and brainstem. Very dense GAT-3 immunoreactivity was observed in the medial septum, diagonal band, basal nucleus of Meynert, thalamus, globus pallidus, and substantia nigra. Moderate levels were observed in the subthalamic nucleus, periaqueductal grey, spinal trigeminal and vestibular nuclei. A general light level of staining was observed in the remainder of the brainstem regions, and very light staining was observed in the caudate nucleus and putamen. Electron microscopy showed that GAT-3 immunoreactivity was present in cell bodies with light cytoplasm and dense bundles of glial filaments, and features of astrocytes. Large numbers of astrocytic processes were also labeled in the neuropil. The cell bodies and processes of neurons were unlabeled. Further study is necessary to elucidate GAT-3 expression in neurological conditions, including hyperalgesia and Parkinson's disease.     

Peptide YY-like immunoreactivity in the central nervous system of the rat

The concentration of peptide YY (PYY)-like immunoreactivity in rat brain and spinal cord was determined by radioimmunoassay. The highest concentrations were found in the cervical spinal cord (18.1 +/- 1.3 ng/g, mean +/- S.E.M.) and in the medulla oblongata (16.3 +/- 1.5 ng/g). Lower amounts were found in the pons and in the hypothalamus. Chromatographic analysis of the PYY-like immunoreactivity from various regions of the brain revealed 95% of the immunoreactive material to be indistinguishable from synthetic porcine PYY. PYY-immunoreactive nerve cell bodies could be demonstrated by immunocytochemistry in the medulla oblongata of colchicine-treated rats, the largest group of cells being found in the midline area between and partly in the raphe pontis and obscurus nuclei. Another large group of immunoreactive cells was detected more laterally in the medial parts of the gigantocellular reticular nucleus. A few cells, finally, were seen in the dorsal parts of the medulla, including the nucleus of the solitary tract. Varicose nerve fibers displaying PYY immunoreactivity were observed in many parts of the hypothalamus, pons, medulla and spinal cord.