The Intrinsic Organization of the Ventroposterolateral Nucleus and Related Reticular Thalamic Nucleus of the Rat: A Double-Labeling Ultrastructural Investigation with γ-Aminobutyric Acid Immunogold Staining and Lectin-Conjugated Horseradish Peroxidase

An electron-microscopic investigation of the synaptic organization of the rat's ventroposterolateral nucleus (VPL) and of a reticular thalamic nucleus (RTN) area related to somatosensory thalamic nucleus was performed. In a group of 11 rats, wheatgerm agglutinin conjugated to horseradish peroxidase (WGA:HRP) was injected either in the first somatosensory area of cortex (SI) or in the dorsal column nuclei (DCN). The retrogradely and/or anterogradely transported enzyme was visualized using paraphenylenediamine-pyrocatechol (PPD-PC) as substrate. In a second series of six experiments, an immunocytochemical procedure using a specific anti-γ-aminobutyric acid (anti-GABA) was employed. Postembedding localization of GABA was performed for ultrastructural observation by means of the colloidal gold immunostaining procedure. Thin sections of recognized VPL and RTN areas from WGA:HRP-injected animals were further processed for immunocytochemistry in order to localize simultaneously, at the electron-microscopic level, the transported enzyme and GABA.

The results obtained with this procedure demonstrated that HRP-labeled terminals from DCN contacted the soma and proximal dendrites of VPL neurons, while the terminals labeled after SI cortical injections were predominantly localized to the distal portion of the dendrites. The same cortical injection also determined the presence of labeled synaptic boutons contacting the soma, and both proximal and distal dendrites of RTN neurons. GABA-immunolabeled terminals were observed in VPL in a number larger than those observed with other methods, since not only typical F terminals were labeled but also terminals containing round and/or pleomorphic vesicles. GABA-ergic terminals contacted the soma and the proximal and distal dendrites of VPL neurons, while in RTN cells they made synaptic contact mainly with the soma and proximal dendrites. In the double-labeling experiments, terminals containing both HRP and specific immunogold GABA staining were never observed.

The present data provide a direct demonstration of the presence of a strong inhibitory input from RTN upon VPL neurons and of the existence of autoinhibition within RTN neurons.     

The Corticocuneate Pathway in the Cat: Relations among Terminal Distribution Patterns,Cytoarchitecture, and Single Neuron Functional Properties

A combined anatomical and physiological strategy was used to investigate the organization of the corticocuneate pathway in the cat. The distribution of the corticocuneate projection was mapped by means of the anterograde horseradish peroxidase (HRP) labeling technique and correlated with the nuclear cytoarchitecture in Nissl and Golgi material, the distribution of retrogradely labeled relay cells after HRP injections in the ventrobasal complex of the thalamus, and the topographic organization derived from single-and multiunit recordings in the decerebrate, unanesthetized cat. This approach provided details about the arrangement of the corticocuneate pathway that were not available from previous studies with anterograde degeneration methods.

On the basis of cytoarchitectonic and connectional features, a number of subdivisions are identified in the cuneate nucleus, each of which is associated with characteristic functional properties. In agreement with previous studies, it is found that a large portion of the cuneate nucleus, the middle dorsal part (MCd), is exclusively devoted to the representation of cutaneous receptive fields on the digits. This “core” region contains more thalamic projecting neurons than any other subdivision of the cuneate nucleus. A topographic arrangement also exists in the subdivisions of the rostral cuneate and of the nuclear region ventral to MCd, although in these, receptive fields are larger and predominantly, but not exclusively, related to deep receptors and involve the arm, shoulder, and trunk.

Observations on corticocuneate projections were based on injections, mainly focused on functional subdivisions of the primary somatosensory cortex (SI) as described by McKenna et al (1981). Although cortical projections are mainly to cuneate regions other than its core, a significant proportion of fibers from the region of SI where the digits are represented (particularly area 3b) do project to the MCd region of the cuneate nucleus. Similarly, nuclear areas associated with receptive fields on the arm and trunk are labeled after injection in SI arm and trunk regions, respectively. Thus, a close topographic relationship appears to exist between the somatosensory cortex and cuneate regions related to the same body representation, although nuclear regions in which receptive fields on the neck area are represented receive very sparse or no detectable cortical projections even when the injection of the tracer involves the entire sensorimotor cortex. The topographic arrangement of SI projections upon the cuneate nucleus suggests that a similar pattern exists in both structures with regard to the relative representations of distal versus proximal and deep versus cutaneous receptive fields (e.g., “core” vs. “shell” organization), and that cuneate regions preferentially related to either of these classes of receptive fields receive direct connections from the corresponding regions in SI.

A comparison of the results from cats with tracer injections in areas 4 and 3b reveals that the projections from the former is denser than that arising from the latter and that their territories of termination largely overlap in the ventral portions of the cuneate nucleus. However, cortical projections to MCd may be derived from the somatosensory cortex with no contribution from area 4. The demonstration of the relative selectivity of cortical projections from different cytoarchitectonic and functional cortical areas to cuneate regions identified here provides a structural basis for the elucidation of the physiological and behavioral observations, particularly on cortical modulation of somatosensory transmission during movements.     

Differences in the Neurons That Project from the Dorsal Column Nuclei to the Diencephalon,Pretectum, and Tectum in the Cat

The dorsal column nuclei (DCN) project to a number of targets in the nervous system besides the ventroposterolateral nucleus (VPL) of the thalamus. Recent evidence obtained using double-labeling techniques indicates that DCN's diencephalic-projecting neurons differ in their location and morphology from those that project to some of its other targets, such as the cerebellum and tectum. The purpose of the present study was to characterize anatomically the DCN neurons that project another of DCN's targets, the pretectum, and to determine if any of these neurons have collateral projections to the tectum or diencephalon.

The projections were studied using two double-labeling methods. One method made use of either tritiated inactivated horseradish peroxidase ([³H]apoHRP) or tritiated N-acetyl wheatgerm agglutinin ([³H]WGA) as a marker and HRP or WGA conjugated to HRP. The other method made use of the dyes Fast Blue and Nuclear Yellow. In each cat, one marker was injected into the DCN-recipient portions of the pretectum, tectum, or diencephalon, and the other marker was injected into another of these three targets.

Neurons labeled by pretectal or tectal injections were of all sizes, fusiform and multipolar in shape, and similarly located. They were scattered through the rostral zone of DCN, but were distributed at the periphery of and at the junction between the gracile and cuneate nuclei in DCN's middle and caudal zones.

In contrast to the pretectal-and tectal-labeled neurons, neurons labeled by diencephalic injections were round and large. They were found throughout the DCN complex, but were concentrated in DCN's middle and caudal zones. When both the pretectum and diencephalon were injected in the same cat, the two groups of neurons occupied similar locations in the rostral zone, but were distinct in the middle and caudal zones, with the pretectal-projecting neurons surrounding the clusters of diencephalic-projecting neurons. Very few neurons were double-labeled.

These results demonstrate that the projections to the pretectum, tectum, and diencephalon originate from different populations of neurons within specific domains in DCN. When these results are compared with the results of electrophysiological and other anatomical studies, it appears that the pretectal- and tectal-projecting neurons may be part of a previously unrecognized system originating in DCN. In contrast with the well-known lemniscal system, recognized for its function in tactile discrimination, and composed of DCN's VPL-projecting neurons together with VPL's projections to the cerebral cortex, this other system may serve some role in the regulation of posture or the coordination of movement.     

Cerebellar projections to the somatic pretectum in the cat

Neurons in the somatic pretectum receive input from the dorsal column nuclei (DCN) and project to a comparable "somatic" portion of the dorsal accessory nucleus of the inferior olive (DAO). This somatic DAO is reciprocally connected with the anterior interpositus nucleus of the cerebellum. One question that arises is whether this circuitry is further controlled by an output specifically from the anterior interpositus nucleus to the somatic pretectum. Wheatgerm agglutinin conjugated to horseradish peroxidase was injected into various parts of the cat pretectum. Injection sites were interpreted as including the somatic pretectum if neurons in the DCN were retrogradely labeled and if anterograde terminal labeling occurred in somatic DAO. The locations of retrogradely labeled neurons within the deep cerebellar nuclei were then compared in cases in which the injection sites included or excluded the somatic pretectum. In all cases in which the injection site included the somatic pretectum, retrogradely labeled neurons were observed in the anterior interpositus nucleus as well as in the lateral cerebellar nuclei. In some of these cases, neurons in the posterior interpositus and medial nuclei were also labeled. In contrast, in cases in which the pretectal injection site was located outside or at the border of the somatic pretectum, retrogradely labeled neurons were observed only in the lateral, posterior interpositus, and medial nuclei. Thus, the somatic pretectum appears to receive input primarily from neurons in the anterior interpositus nucleus, along with some input from neurons in the lateral nucleus. These results provide additional evidence for a pathway through the DCN in which sequentially processed somatic information has access to and is modulated by cerebellar circuitry. The existence of such a pathway supports the conclusion that neurons in the DCN convey somatic information important not only for cutaneous, kinesthestic, and other bodily sensations, but also for the control of movement.     

Cytology of large neurons in the guinea pig dorsal cochlear nucleus contacting the inferior colliculus

Large neurons in the dorsal cochlear nucleus of the guinea pig which project to the inferior colliculus were identified after injections of the neural tracer WGA-HRP. Retrograde labelled cells (pyramidal and giant neurons) in the dorsal cochlear nucleus were glycine and GABA immunonegative and showed a similar ultrastructure. Between 30 and 60% of their perimeter was covered by axo-somatic boutons, most of which (>50%) contained pleomorphic synaptic vesicles. Other boutons (about 40% of total) contained flat vesicles and few (5-6%) contained round vesicles, a characteristic of the excitatory cells innervating the inferior colliculus. Immunogold-cytochemistry, coupled to silver intensification, showed that more than 50% of axo-somatic pleomorphic boutons and over 90% of boutons containing flat and pleomorphic vesicles store glycine. Rare WGA-HRP labelled axo-somatic boutons containing flat-pleomorphic vesicles were seen on pyramidal and giant neurons. This suggests that a few inhibitory collicular terminals contact the excitatory large neurons in the dorsal cochlear nucleus.     

The synaptic relationships between the primary afferent terminals and the cuneo-thalamic relay neurons in the rat cuneate nucleus

In order to establish the synaptic relationship between the primary afferent terminals and the cuneothalamic relay neurons in the cuneate nucleus, the combined retrograde transport of horseradish peroxidase (HRP) and experimental degeneration have been applied in the young adult albino rats. 10 to 30% HRP was injected contralaterally (0.5 microliter) in the ventrobasal thalamic nucleus and multiple dorsal rhizotomies (C5 to T1) in the cervicothoracic dorsal roots were performed on the side ipsilateral to the cuneate nucleus. The results showed that: The cuneo-thalamic relay (CTN) neurons were the major neuronal type of the nucleus. More than 55% of neurons have been labelled. These neurons were 18-30 micron X 15-25 micron in sizes. They distributed in the whole rostrocaudal extent of the nucleus, particularly dense in the middle portion. The cells varied from round, oval, spindle to multipolar in shapes. They were rich in cytoplasmic organelles and had well-developed roughed endoplasmic reticulum. Their nucleus was either centrally or eccentrically located and was rather regular. The HRP-positive granules were randomly distribute in the perikaryon, dendrites and initial segment of the axons; At least three types of the experimental degeneration of the primary afferent terminals (PAT) were observed in the cuneate nucleus two to three days after dorsal rhizotomy, namely, electron-dense, granular and neurofilamentous. These PAT were mostly large and contained round vesicles. They were commonly found within synaptic complex, in which they were presynaptic to dendrites of various sizes, and were themselves postsynaptic to smaller axon terminals containing flattened vesicles. Degenerating PAT forming isolated synapses were less commonly seen; The PAT in the synaptic complex were directly presynaptic to the dendrites originating from the CTN neurons. The dendrites forming PAT-CTN synases were of large and medium-sized. The PAT did not form direct axo-somatic synapses with the somata of CTN or of any other cell types in the cuneate nucleus.     

Effect of Microstimulation of Movement-Evoking Cortical Foci on the Activity of Neurons on the Dorsal Column Nuclei

Cortical foci in which stimulation produced movement in either the forelimb or hindlimb were isolated in rats. In each experiment, two foci were selected: one for movement in the forelimb, and the other in the hindlimb. Stimulation was subsequently reduced in order to avoid eliciting a movement, and the effects of this stimulation on activity of gracile and cuneate neurons were examined. Both excitation and inhibition were observed and were found to be arranged in a somatotopic manner. Excitation was almost exclusively obtained when the receptive field (RF) of a given neuron corresponded to the body surfaces overlying the joints involved in the cortically evoked movement. A high percentage of neurons with RFs on body surfaces corresponding to, or adjacent to, the region of cortically induced movement were inhibited, while the activity of neurons with RFs distant to the site of movement was seldom modified. These results suggest that cortical influences exerted on the dorsal column nuclei (DCN) in rats are organized in a somatotopic manner.     

Identification of tuberculo-ventral neurons in the polymorphic layer of the rat dorsal cochlear nucleus

The tuberculo-ventral tract represents a short nervous circuit within the auditory cochlear nuclei. Tuberculo-ventral neurons of the dorsal cochlear nucleus send isofrequency inhibitory inputs to bushy cells of the ventral cochlear nucleus. Injection of wheat germ agglutinin conjugated to horseradish peroxidase into the rat ventral cochlear nucleus, labelled tuberculo-ventral neurons retrogradely in the deep polymorphic layer of the ipsilateral dorsal cochlear nucleus. Five to 20% of the perimeter of these cells was covered by synaptic boutons, most of which contained flat and pleomorphic vesicles. These boutons contained glycine and sometimes GABA. Occasional small axo-somatic boutons contained round vesicles and were immunonegative for both glycine and GABA. This study shows that the synaptic profile of tuberculo-ventral neurons is different from that of other medium-size glycinergic neurons within the polymorphic layer or more superficial regions of the dorsal cochlear nucleus like cartwheel neurons. In fact the latter mostly receive boutons that contain pleomorphic vesicles.     

Synaptic terminals in the ventroposterolateral nucleus of the thalamus from neurons in the dorsal column and lateral cervical nuclei: an electron microscopic study in the cat

Brain Cell Biology - The afferent fibres to the ventroposterolateral nucleus (VPL) of the contralateral thalamus from neurons in the dorsal column nuclei (DCN) and the lateral cervical nucleus...     

NADPH-diaphorase, nitric oxide synthase, and tyrosine hydroxylase in the diencephalon of the Rhodeus sericeus (Cyprynidae:Teleostei)

The presence and distribution of nitric oxide synthase (NOS)-like neurons as well as tyrosine hydroxylase-immunoreactive (TH) neurons was studied in the diencephalon of the cypriniform teleost Rhodeus sericeus. The anatomical relationships between tyrosine hydroxylase (TH)- and nitric oxide synthase (NOS)-containing cells were visualized both by NOS-immunohistochemistry and NADPH-histochemistry. Immunohistochemical labeling and morphological studies were performed on the same sections. The results reported in this paper show that both a NOS and TH activity are present in the preoptic region, posterior tuberculum, paraventricular organ and hypothalamus of R. sericeus. Putative nitrergic neurons were identified in all major hypophysiotrophic nuclei of the R. sericeus brain using both NADPH-d histochemistry and nNOS immunohistochemistry. In the preoptic region, nitrergic neurons were found in both the parvocellular and the magnocellular nuclei. Within these nuclei, the distribution of NADPH-d reactivity was similar to that of nNOS immunoreactivity. However, we found no evidence of colocalization of NADPH-d and nNOS in consecutive sections. NOS- and TH-containing neurons were observed in all the nuclei under study (hypothalamus, posterior tuberculum, ventral thalamus) and telencephalon (preoptic region), although most neurons showing the coexistence of both substances were mainly located in the preoptic nucleus and hypothalamus, some labelled neurons were found in the posterior tuberculum. Most of the cerebrospinalliquor-contacting cells (LCNs) in diencephalic periventricular area of R. sericeus were TH-immunoreactive. Also, a large number ofnitrergic small LCNs distributed throughout the third ventricle were observed in these regions. The data obtained supports the existence of a nitrergic circumventricular system in teleost. LCNs in R. sericeus are thought to be involved in osmoregulatory functions as osmosensitive neurons. Due to their chemical properties, NO produced by these cells might play an important role in the maintenance and regulation of CSF homeostasis through the modulation of cerebral blood flow.     

溴化乙锭诱导的短暂脱髓鞘增强PRV的逆行转导效率 *

目的 改进伪狂犬病病毒(Pseudorabies virus,PRV)的注射方法,通过溴化乙锭(ethidium bromide,EB)诱导的短暂脱髓鞘提高PRV的转导效率。方法 选用18只正常成年Wistar大鼠,随机分为肌肉组、NS组和EB组(n=6)。肌肉组将2μl滴度为2×10 ⁹的PRV工具病毒注射到胫骨前肌和腓肠肌上;NS组将2μl生理盐水(normal saline,NS)注射到坐骨神经上,1周后相同位置注射2μl滴度为2×10 ⁹的PRV;EB组将2μl 0.1%的EB注射到坐骨神经上,1周后相同位置注射2μl滴度为2×10 ⁹的PRV。大鼠注射PRV 5天后,灌流取材并制作冰冻切片,观察各级神经元的感染情况。 结果 EB组大鼠L4-L5脊髓前角神经元和背根神经节(DRG)、T8脊髓中间神经元、C4脊髓中间神经元、延髓、中脑、大脑皮层均有大量神经元被PRV标记。肌肉组和NS组各级神经元仅有少量被PRV标记。结论 EB诱导坐骨神经脱髓鞘后能够显著提高PRV的逆行转导效率。     

GABAergic input of cholecystokinin-immunoreactive neurons in the hilar region of the rat hippocampus

Summary Double immunostaining was performed for electron microscopy to analyze the synaptic connections between glutamate decarboxylase (GAD)-immunoreactive axons and cholecystokinin (CCK)-immunoreactive neurons in the hilar region of the rat hippocampal formation. Following immunostaining for CCK, the diaminobenzidine (DAB) reaction product was silver-intensified and gold-substituted. In a subsequent second immunostaining for GAD, the immunoreactive elements were labeled using a single DAB reaction. Electron microscopic analysis of the double-stained Vibratome sections demonstrated that the single DAB-labeled GAD-immunoreactive boutons form symmetrical synaptic connections on the soma and primary dendrites of the DAB-gold-labeled CCK-immunoreactive neurons.     

Immunocytochemical identification of GABAergic neurons in the main olfactory bulb of the rat

With the aid of a sheep antiserum against rat brain glutamate decarboxylase (GAD), the endogenous marker for GABAergic neurons, we have labeled immunocytochemically various types of nerve cells in the main olfactory bulb of rats, with and without topic injections of colchicine. The peroxidase-antiperoxidase procedure was applied to floating Vibratome and frozen sections. A large part of the periglomerular cell population and practically all granule cells in the deep layers contain GAD-like immunoreactivity in untreated rats, while tufted and mitral cells (the projection neurons) are unstained. This observation confirms a previous study with a rabbit antiserum against mouse brain GAD, which suggested that GABAergic neurons with presynaptic dendrites contain high somatal concentrations of GAD. We show, however, that immunostaining of granule cell bodies decreases progressively from the internal plexiform layer to the deep portion of the granule cell layer. Many cell processes in the glomeruli are densely stained. They presumably represent synaptic gemmules of the numerous GAD-positive periglomerular cells, which thus could provide initial, inhibitory modulation of the afferent input. In the external plexiform layer immunostaining of the neuropil is substantially denser in the superficial half than in the deep half. This may reflect a corresponding gradient of inhibition related to unequal frequency of occurrence of synaptic gemmules of granule cell dendrites. Alternatively such a graded immunostaining of cell processes could be related to the corresponding gradient in the density of immunostaining of granule cell bodies in the deep layers, in accordance with recent data indicating that superficial and deep granule cells project their ascending dendrites respectively to superficial and deep portions of the external plexiform layer. Furthermore, we have demonstrated the presence of additional classes of GAD-positive neurons, microneurons in the external plexiform layer, small neurons in the periglomerular region, the external plexiform layer, the mitral cell layer, the internal plexiform layer, and medium-size neurons in the granule layer and the white matter. The small- and medium-size GAD-positive neurons appear weakly immunoreactive in untreated rats, but become densely stained after topic colchicine injection. Such cells presumably lack presynaptic dendrites and may correspond to different types of short axon cells demonstrated by the Golgi method.(ABSTRACT TRUNCATED AT 400 WORDS)     

Innervation of the dermatomes in the neck of the mouse

The dorsal rami of the cervical and thoracic spinal nerves were investigated using both the in situ cholinesterase staining technique and cholinesterase staining on serial sections of plastic-embedded embryos. In most cases only the dorsal rami of the 2nd to 5th cervical spinal nerve possess cutaneous branches. The area innervated by the cutaneous branch of the dorsal ramus of the 5th spinal nerve borders on an area innervated by the cutaneous branch of the dorsal ramus of the 1st thoracic spinal nerve. The dorsal rami of the cervical spinal nerves 6-8 show no cutaneous branches. Therefore the gap in the series of the dorsal cutaneous branches is due only to the middle part of the nerves of the brachial plexus, which range from the 5th cervical nerve to the 1st thoracic nerve.     

GABAergic input of cholecystokinin-immunoreactive neurons in the hilar region of the rat hippocampus. An electron microscopic double immunostaining study

Double immunostaining was performed for electron microscopy to analyze the synaptic connections between glutamate decarboxylase (GAD)-immunoreactive axons and cholecystokinin (CCK)-immunoreactive neurons in the hilar region of the rat hippocampal formation. Following immunostaining for CCK, the diaminobenzidine (DAB) reaction product was silver-intensified and gold-substituted. In a subsequent second immunostaining for GAD, the immunoreactive elements were labeled using a single DAB reaction. Electron microscopic analysis of the double-stained Vibratome sections demonstrated that the single DAB-labeled GAD-immunoreactive boutons form symmetrical synaptic connections on the soma and primary dendrites of the DAB-gold-labeled CCK-immunoreactive neurons.     

Anatomical changes of the GABAergic system in the inferior colliculus of the genetically epilepsy-prone rat

The number of GABAergic neurons as determined by GAD immunocytochemistry and total neurons as determined from Nissl preparations were counted and classified at the light microscopic level in the inferior colliculus (IC) of the genetically epilepsy prone rat (GEPR) and the non-epileptic Sprague-Dawley (SD) strain of rat. GAD-positive neurons are abundant in the IC and a significant increase in the number of GAD-positive neurons occurs in the GEPR as compared to the SD in all three subdivisions. However, the most pronounced difference occurs in the ventral lateral portion of the central nucleus, where there is a selective increase in the small (200%) and medium-sized (90%) GABAergic somata (10-15 microns in diameter and 15-25 microns in diameter, respectively). As determined from Nissl preparations an increase in total numbers of neurons also occurs. Thus, a 100% increase in the number of small neurons and a 30% increase in the number of medium-sized neurons occur in the adult GEPR as compared to the SD rat. A statistically significant increase in the numbers of small neurons also occurred in the IC of the young GEPR. At 4 days of age, a 55% increase in the number of small neurons was found, and at 10 days of age this increase was 105%. The numbers of the medium and large neurons were similar in the older group of rats. These data suggest that the increase in cell number observed in the adult GEPR is not compensatory to the seizure activity, but may either be genetically programmed or be a failure of cell death. Based on other studies of genetic models of epilepsy, we propose that the additional GABAergic neurons may disinhibit excitatory projection neurons in the IC.     

Distinct origin of GABA-ergic neurons in forebrain of man, nonhuman primates and lower mammals

In this mini-review we present recent data about origin of GABA-ergic (gama-aminobutyric acid) neurons in the mammalian forebrain, including the diencephalon and telencephalon. The interest in GABA-ergic neurons, which in cerebral cortex mostly correspond to local circuit neurons (interneurons), has increased in the past decade. Many studies have shown that in lower mammals all hippocampal and almost all neo-cortical GABA-ergic neurons are born in the specific region named ganglionic eminence, and not locally in proliferative layers all around telencephalic vesicle. The ganglionic eminence, that represents a region with thick proliferative-subventricular layer in the ventral (basal) part of telencephalon, was classically thought to give neurons to basal ganglia and septal nuclei, whereas proliferative layers of dorsal telencephalon give neurons to cerebral cortex including hippocampus. It was thought that neurons migrate from proliferative layer to their target region following a radial orientation. However, data in lower mammals showed that this is the case only for glutamatergic principal cells, i.e. projection neurons. GABA-ergic neurons use long distance tangentional migration, parallel to pial surface to reach, from ganglionic eminence, their targeting layer in the cerebral cortex. Especially intriguing, but frequently neglecting, several studies suggest that mammalian evolution might use different developmental rules to provide GABA-ergic neurons to an expending brain. In this review we focus on specific events underlying GABA-ergic neuron development in human and non-human primates. Disturbances of the GABAergic network are found in many neurological and psychiatric disorders, some of them might result from altered production or migration of these neurons during development. Therefore, it is crucial to understand human-specific mechanisms that regulate the development of GABA-ergic neurons.     

Electron microscopic analysis of tyrosine hydroxylase, dopamine-beta-hydroxylase and phenylethanolamine-N-methyltransferase immunoreactive innervation of the hypothalamic paraventricular nucleus in the rat

The catecholaminergic innervation of the hypothalamic paraventricular nucleus (PVN) of the rat was studied by preembedding immunocytochemical methods utilizing specific antibodies which were generated against catecholamine synthesizing enzymes. Phenylethanolamine-N-methyltransferase (PNMT)-immunoreactive terminals contained 80-120 nm dense core granules and 30-50 nm clear synaptic vesicles. The labeled boutons terminated on cell bodies and dendrites of both parvo- and magnocellular neurons of PVN via asymmetric synapses. The parvocellular subnuclei received a more intense adrenergic innervation than did the magnocellular regions of the nucleus. Dopamine-beta-hydroxylase (DBH)-immunopositive axons were most numerous in the periventricular zone and the medial parvocellular subnucleus of PVN. Labeled terminal boutons contained 70-100 nm dense granules and clusters of spherical, electron lucent vesicles. Dendrites, perikarya and spinous structures of paraventricular neurons were observed to be the postsynaptic targets of DBH axon terminals. These asymmetric synapses frequently exhibited subsynaptic dense bodies. Paraventricular neurons did not demonstrate either PNMT or DBH immunoreactivity. The fibers present within the nucleus which contained these enzymes are considered to represent extrinsic afferent connections to neurons of the PVN. Tyrosine hydroxylase (TH)-immunoreactivity was found both in neurons and neuronal processes within the PVN. In TH-cells, the immunolabel was associated with rough endoplasmic reticulum, free ribosomes and 70-120 nm dense granules. Occasionally, nematosome-like bodies and cilia were observed in the TH-perikarya. Unlabeled axons established en passant and bouton terminaux type synapses with these TH-immunopositive cells. TH-immunoreactive axons terminated on cell bodies as well as somatic and dendritic spines of paraventricular parvocellular neurons. TH-containing axons were observed to deeply invaginate into both dendrites and perikarya of magnocellular neurons. These observations provide ultrastructural evidence for the participation of central catecholaminergic neuronal systems in the regulation of the different neuronal and neuroendocrine functions which have been related to hypothalamic paraventricular neurons.     

Light and electron-microscopic study of leucine enkephalin immunoreactivity in the cat claustrum

The claustrum is a complex telencephalic structure owing to its reciprocal connectivity with most—if not all—cortical areas. However, there is a paucity of data in the literature concerning its histochemical components, including opioid peptide neurotransmitters. The aim of the present study was to examine the morphology, distribution and ultrastructure of leucine-enkephalin-immunoreactive (Leu-enk-ir) neurons and fibers in the dorsal claustrum (DC) of the cat. Seven healthy, adult male and female cats were used in our study. All animals received humane care. They were irreversibly anesthetized and transcardially perfused with fixative. Brains were removed, postfixed, blocked and sectioned. Sections were incubated with polyclonal anti-Leu-enk antibodies using the Avidin–Biotin–Peroxidase Complex method. Leu-enk-ir neurons and fibers were distributed throughout the DC. Some of the neurons were lightly-stained, while others were darkly-stained. Light-microscopically, they varied in shape: oval, fusiform, multipolar and irregular. With regard to size, they were categorized as small (15?μm or less in diameter), medium (16–20?μm in diameter) and large (21?μm or more in diameter). No specific pattern of regional distribution was found. On the electron microscope level, immunoproduct was observed in neurons, dendrites and terminal boutons. Different types of Leu-enk-ir neurons differ in their ultrastructural features, including two types of synaptic boutons. No gender-specific features were observed. In conclusion, it is our hope that our study will serve to contribute to a better understanding of the functional neuroanatomy of the DC in the cat, and that it can be extrapolated and applied to other mammals, including humans.