Clear Synaptic Vesicles Research Articles

Somatostatin-immunoreactive amacrine cells of chicken retina: Retinal mosaic, ultrastructural features, and light-driven variations in peptide metabolism

Somatostatin-like immunoreactive amacrine cells of the chicken retina have been characterized by immunohistochemistry at the light and electron microscope levels. The cell bodies were set back from the junction of the inner nuclear and inner plexiform layers, and prominent fibre plexuses were found in sublaminas 1 and 3–5 of the inner plexiform layer. The cells were distributed across the retinal surface with a centroperipheral gradient of cell density. Locally, the cells were organized in a non-random mosaic. Ultrastructurally, immunohistochemical reaction product was found throughout the cytoplasm of the cell bodies, particularly associated with membranous structures, including the cytoplasmic surfaces of the Golgi apparatus, and within large dense-core vesicles. In dendritic varicosities in the inner plexiform layer, reaction product was associated with the external surfaces of small, clear synaptic vesicles. The synaptic relationships of the somatostatin-immunoreactive terminals in sublamina 1 were distinct from those in sublaminas 3–5. Those in sublamina 1 received input predominantly, possibly exclusively, from bipolar cells. Feedback synapses onto bipolar terminals or to the other amacrine cell process at a synaptic dyad were observed. In sublaminas 3–5, input came predominantly, possibly exclusively, from other, non-immunoreactive amacrine cells, and output was primarily onto other amacrine cells. No synaptic contacts with ganglion cells or with other somatostatin-immunoreactive amacrine cells were identified. Changes in levels of somatostatin-like immunoreactivity in retinas of chicks kept on 12:12 light:dark cycles were detected by radioimmunoassay, and by light and electron microscopic immunohistochemistry. Levels of retinal somatostatin-like immunoreactivity increased in the light and decreased in the dark. The changes appear to be light-driven rather than circadian, since with prolonged exposure to light or dark, the levels of somatostatin-like immunoreactivity continued to increase or decrease until plateaus were reached. The light-driven change in levels of somatostatin-like immunoreactivity may be related to the predominance of bipolar input to the immunoreactive processes in sublamina 1 of the inner plexiform layer. The reduction in peptide levels in the dark may indicate greater release of somatostatin-like immunoreactivity from the amacrine cells in the dark, resulting in an inability of peptide synthesis to keep pace with breakdown. In the light, release of somatostatin-like immunoreactivity may be lower, leading to a net synthesis of peptide.

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.

Morphology of physiologically identified slowly adapting lung stretch receptor afferents stained with intra-axonal horseradish peroxidase in the nucleus of the tractus solitarius of the cat. II. An ultrastructural analysis.

The nucleus of the tractus solitarius is a site for termination of primary afferents originating from a variety of visceral receptors. The localization of bouton terminals of slowly adapting lung stretch (SAR) afferent fibers originating from the tracheobronchial tree have been described in the companion paper (Kalia and Richter, '85). The most conspicuous finding regarding the location of SAR terminals is that they are concentrated within specific subnuclear groups of the nucleus of the tractus solitarius (nTS) and are distributed widely in the rostrocaudal plane of the medulla oblongata. These light microscopic features have provided us with valuable information with regard to the organization of visceral afferents in the central nervous system. The synaptic profiles formed by the 476 bouton terminals of these HRP-labeled afferents have been described in this paper in serial thin sections. All of the bouton terminals examined under the electron microscope were found to contain round synaptic vesicles. Synaptic boutons (1.0-3.0 microns in diameter) were usually of the en passant variety and made contact with different structures depending upon the subnucleus which was examined. In the ventral (v) and the ventrolateral (vl) subnuclei of the nTS, asymmetrical (type I) synaptic contacts containing round, clear synaptic vesicles of 35-50 microns in diameter were found and these contacts were made with (1) the soma of cell bodies located in that subnucleus; (2) spiny dendrites in that nucleus; (3) vesicle-containing axon terminals that were presynaptic to the HRP-labeled bouton terminal; and (4) vesicle-containing dendrites in which the HRP profile was presynaptically located. The terminal axon remained myelinated till the last 1 micron before the bouton terminal was formed. There was no distinct, unmyelinated portion of the terminal axon. The synaptic bouton received axon-axonal synapses from unlabeled bouton terminals containing round, clear vesicles. This is the first report of the localization of these afferent fibers as well as of the regional variations in the ultrastructure of boutons of physiologically identified terminals. It appears likely that the lung stretch afferent fibers, by having axon-axonal as well as axon-somatic contact in the ventral, ventrolateral, and intermediate subnuclei of the nTS, can interact in a variety of different ways in this region. The significance of these features in relation to the precise influence of respiratory afferents on central respiratory mechanisms needs to be evaluated further.

Neurons dissociated from rat myenteric plexus retain differentiated properties when grown in cell culture: I. Morphological properties and immunocytochemical localization of transmitter candidates

We have developed procedures for dissociating neurons from the myenteric plexus of the small intestine of newborn rats and for growing those neurons in cell cultures for up to 3 months. Neurons in these cultures retain many of the differentiated properties of myenteric neurons in vivo. This is the first of a series of 3 papers describing those properties. In this paper, we describe the morphology of cultured neurons that we have observed with light and electron microscopy; we also describe the patterns of straining observed when immunocytochemical techniques were used to localize neurotransmitter candidates in the cultured neurons. Intracellular injections of a fluorescent dye, Lucifer yellow, revealed that many of the cultured neurons had morphologies similar to those of myenteric neurons in vivo. When thin sections of cultures were viewed in an electron microscope, many neurons were observed to have numerous small (40–60 nm), clear synaptic vesicles and/or large (80–150 nm), opaque-cored (p-type) vesicles. Synaptic profiles were most often observed on neuronal somata. Neurons containing immunoreactive serotonin, substance P, somatostatin, enkephalin, bombesin and gastrin/cholecystokinin were observed in about the same proportions as they occur in the intact myenteric plexus. Neurons containing immunoreactive vasoactive intestinal polypeptide were found in higher numbers than reported in vivo. Neurons containing immunoreactive neurotensin, secretin and glutamate decarboxylase were not observed. An antiserum directed against choline acetyltransferase stained 40–50% of the neurons. We conclude that myenteric neurons continue to express much of their normal differentiated properties even when they are removed from the gut, dissociated into a suspension of single cells and grown in culture. Such cultures will be useful for correlating the morphological, biophysical, pharmacological and synaptic properties of individual myenteric neurons and for testing the ability of altered environmental conditions to change those properties.

The mormyrid brainstem—III. Ultrastructure and synaptic organization of the medullary “pacemaker” nucleus

The ultrastructure and synaptic organization of the presumed medullary pacemaker nucleus, nucleusc of the weakly electric mormyrid fish, Gnathonemus petersii has been investigated. Nucleusc consists of about 12–15 small (20–25 μm) neurones (P-cells), which form a group situated ventrally to the medullary relay nucleus and embedded in a neuropil of myelinated fibres and dendritic processes. The P-cells often exhibit an enhanced electron density of their cytoplasm and dendroplasm. They possess several dendrites of different diameter, a short, thin axon initial segment and a thickly myelinated axon running in dorsal direction. The pacemaker neurons are interconnected by complex electronic coupling, established by somatosomatic, dendrosomatic and dendrodendritic gap junctions. Perikarya and dendrites are frequently interconnected serially by gap junctions; dendrites showed sometimes triadic gap-junction arrangement. It is suggested that this high degree of electrotonic coupling amongst the pacemaker cells represents the first level of the highly ordered synchronization processes which characterize the electric discharge command system of Gnathonemus. Pacemaker cells receive synaptic input from club endings with mixed synapses and from bouton-like terminals with chemical synapses, both of them originating from medium-sized myelinated fibres and contacting mainly neuronal perikarya and dendritic processes. The axon initial segment receives only few synaptic inputs. Bouton-like terminals were found to be of two types according to their vesicle content, namely, boutons with ovoid, clear synaptic vesicles forming Gray type-1 synapses and boutons with pleomorphic clear synaptic vesicles forming Gray type-2 synapses. Different functional roles for the two types of boutons in modulating pacemaker cell activity are suggested.

Immunocytochemical observations of corticotropin-releasing-factor-containing neurons in the rat hypothalamus with special reference to neuronal communication.

Synapses between neurons with corticotropin-releasing-factor-(CRF)-like immunoreactivities and other immunonegative neurons in the hypothalamus of colchicine-treated rats, especially in the paraventricular nucleus (PVN) and the supraoptic nucleus (SON) were observed by immunocytochemistry using CRF antiserum. The immunoreactive nerve cell bodies and fibers were numerous in both the PVN and the SON. The CRF-containing neurons had synaptic contacts with immunonegative axon terminals containing a large number of clear synaptic vesicles alone or combined with a few dense-cored vesicles. We also found CRF-like immunoreactive axon terminals making synaptic contacts with other immunonegative neuronal cell bodies and fibers. And since some postsynaptic immunonegative neurons contained many large neurosecretory granules, they are considered to be magnocellular neurosecretory cells. These findings suggest that CRF functions as a neurotransmitter and/or modulator in addition to its function as a hormone.

Ultrastructural study of cholecystokinin-like immunoreactive nerve terminals in the rat nucleus accumbens

The cholecystokinin (CCK)-like immunoreactive nerve terminals were studied in the caudal and medial parts of the rat nucleus accumbens (NA), using the indirect immunoperoxidase technique, at the electron microscopic level. In the labelled axon terminals the immunoprecipitate is localized inside large densecored vesicles which are occasionally present, and surrounds small and medium-sized, round, clear synaptic vesicles. The immunoreactive nerve terminals participate in synapses of both asymmetrical and symmetrical types containing mostly small synaptic vesicles. The asymmetrical synapses are much more numerous and mainly axo-spinous. The symmetrical synapses are less frequent and are axo-dendritic or axo-somatic.

Cartwheel neurons of the dorsal cochlear nucleus: a Golgi-electron microscopic study in rat.

Cartwheel neurons in rat dorsal cochlear nucleus (DCN) were studied by Golgi impregnation-electron microscopy. Usually situated in layers 1-2, cartwheel neurons (10-14 micrometers in mean cell body diameter) have dendritic trees predominantly in layer 1. The dendrites branch at wide angles. Most primary dendrites are short, nontapering, and bear only a few sessile spines. Secondary and tertiary dendrites are short, curved, and spine-laden. The perikaryon forms symmetric synapses with at least two kinds of boutons containing pleomorphic vesicles. The euchromatic nucleus is indented and has an eccentric nucleolus. The cytoplasm shows several small Nissl bodies, a conspicuous Golgi apparatus, and numerous subsurface and cytoplasmic cisterns of endoplasmic reticulum with a narrow lumen, joined by mitochondria in single or multiple assemblies. In primary dendrites mitochondria are situated peripherally, while in distal branches they become ubiquitous and relatively more numerous. Dendritic shafts usually form symmetric synapses with boutons that contain pleomorphic vesicles. The majority of the dendritic spines are provided with a vesiculo-saccular spine apparatus. All dendritic spines have asymmetric synapses. Most of these are formed with varicosities of thin, unmyelinated fibers (presumably axons of granule cells) running parallel to the long axis of the DCN or radially. These varicosities contain round, clear synaptic vesicles. On the initial axon segment few symmetric synapses are present. The axon acquires a thin myelin sheath after a short trajectory. Cartwheel neurons outnumber all other neurons in layers 1-2 (with the exception of granule cells), and presumably correspond to type C cells with thinly myelinated axons described by Lorente de Nó. The axons of these neurons provide a dense plexus in the superficial layers without leaving the DCN. The possible functional role of cartwheel neurons is discussed.

Development of the axon cap neuropil of the Mauthner cell in the goldfish.

Development of the axon cap neuropil of the Mauthner neuron in post-hatching larval goldfish brains was observed electron-microscopically. The axonal initial segment of newly hatched (day-4) larvae is completely covered with synaptic terminals containing clear spherical synaptic vesicles. Profiles of thin terminal axons, the spiral fibers, containing similar synaptic vesicles, rapidly increase in number around the initial segment and form glomerular neuropil similar to the central core of the adult axon cap by day 7. Three types of synapses are formed in the core neuropil. Bouton-type synapses contacting the initial segment are most abundant in day-4 to -14 larvae; they decrease thereafter and are rare on the distal half of the initial segment of day-40 larvae. Asymmetric axo-axonic synapses are commonly observed between spiral fibers in the core neuropil of day-7 to -19 larvae, but become fewer by day 40. Unique symmetrical axo-axonic synapses showing accumulation of synaptic vesicles on either side of apposed membrane thickenings first appear in day-14 core neuropil, gradually increase in number, and become the predominant type in day-40 core neuropil. Thick myelinated axons, which lose their myelin sheaths in the glial cap cell layer, start to penetrate into the axon cap on day 10. They gradually increase in number and form the peripheral part of the axon cap together with the cap dendrites, which finally grow into the axon cap from the axon hillock region of the Mauthner cell by day 40.

The primary afferent projection of the greater petrosal nerve to the solitary complex in the rat, revealed by transganglionic transport of horseradish peroxidase

The primary afferent projection of the greater petrosal nerve (GPN) to the solitary complex was studied following application of horseradish peroxidase (HRP) to the GPN just distal to the geniculate ganglion. Labeled fibers were traced to the most rostral part of the solitary tract. Numerous collaterals entered the solitary complex from its dorsal and lateral aspects, and formed a dense plexus. They terminated in the dorsal half of the medial solitary nucleus at the level of the rostral half of the solitary complex, and in the ventrolateral and commissural nuclei at the level of the caudal half. The densest termination was observed in the medial solitary nucleus. Labeled terminals were found to contain round, clear synaptic vesicles and to make asymmetrical synaptic contacts with dendritic profiles.

Demonstration of electron-dense material in clear synaptic vesicles using cationic ferrocenyl compounds.

Electron-dense material in clear synaptic vesicles in rat cerebral cortex and neuromuscular junctions of frog cutaneous pectoris muscle was demonstrated by using ferrocenyl cationics. Electron-dense spots were usually attached to the inner surface of the vesicular membrane. Control experiments (treatment with Triton X-100 or cetylpyridinium chloride; enzyme digestion with trypsin, hyaluronidase, neuraminidase, sulfatase and beta-glucuronidase) suggested that the electron-dense material is a glycoprotein.

Primary afferent terminals in the nucleus of the solitary tract of the frog: an electron microscopic study.

In the frog solitarius nucleus, primary afferent terminals of the facial and glossopharyngeal-vagal nerves were identified with cobalt labelling and electron microscopy. The labelled terminals were grouped in two main categories, one with small (1-2 micron) and pale terminals, and another with large (3-5 micron) and dark terminals. The small terminals greatly outnumbered the large ones. In addition many terminals intermediate in size and staining reactions were found. All kinds of labelled boutons contained medium-size clear synaptic vesicles, among which dense-core vesicles of the smaller type frequently occurred. The labelled primary afferent terminals established axo-dendritic contacts of the asymmetric type. Close to these contact sites they were themselves very frequently contacted by a profile interpreted as presynaptic in relation to them. Such profiles contained spherical, pleomorphic (including dense-core) or flattened vesicles; a fourth kind was interpreted as presynaptic dendrites. It is concluded that viscerosensory fibres, as opposed to somatosensory fibres, predominantly generate small and lightly stained terminals. It is likely that the effect of synaptic transmission at the solitarius tract terminals is modulated in a very versatile manner by the various presynaptic profiles converging on these terminals.

Ultrastructural studies of neuromuscular junctions in visceral and skeletal muscles of the chaetognath Sagitta setosa.

The ultrastructural characteristics of the neuromuscular junctions were studied in oesophageal (visceral) muscle and in four skeletal muscles of the head and trunk in Sagitta. Three types of neuromuscular junctions were encountered. The first is made up of nerve terminals which synapse with the surface of the muscle fiber, in a deep or in a slight depression. The second is characterized by muscle fiber protrusions that cross the connective tissue and form synapses with nerve endings; in this type, numerous post-junctional membrane folds are noted. In the third type, the synaptic cleft is very large (greater than 0.2 micron) and contains bundles of connective fibers. Nerve endings are partially ensheathed in glial cells; they contain mostly clear synaptic vesicles, though some dense-cored vesicles are noted. In many muscle fibers post-junctional membrane thickenings are also observed. All observed neuromuscular junctions resemble chemical synapses. Chaetognaths thus show a great variety of neuromuscular junction ultrastructure as do for instance Arthropods.

Light- and electron-microscopic immunocytochemistry of glutamic acid decarboxylase (GAD) in the basal hypothalamus: Morphological evidence for neuroendocrine γ -aminobutyrate (GABA)

GABAergic cells and axon terminals were localized in the basal hypothalamus of different species (rat, mouse and cat), by means of an immunocytochemical approach using a specific and well-characterized antiserum to the GABA biosynthetic enzyme, glutamate decarboxylase. Lightmicroscopic visualization was performed with an indirect immunofluorescence method and electron-microscopic observations were made on material with pre-embedding staining and use of the peroxidase-antiperoxidase procedure. At the light-microscopic level, a dense immunofluorescent plexus was observed over both the medial and lateral parts of the external layer of the median eminence. The labelling extended from the rostral part of the median eminence up to the pituitary stalk. Over the subependymal and internal layers only a few immunoreactive dots were visible, except around the blood vessels where they appeared more concentrated. Immunoreactive varicosities could be found following the outlines of the capillary loops and lining tanycyte processes, especially in the median eminance midportion. At the electron-microscopic level, the immunolabelling was exclusively found over neuronal profiles in the median eminence. The latter represented a small fraction of the total number of varicosities visible on the same section. Labelled profiles typically contained numerous small clear synaptic vesicles and only a few or no dense-core vesicles. In the subependymal and internal layers, rare labelled endings were found close to ependymal cells or among transversally cut fibers, respectively. In the palisadic zone, elongated positive boutons were visible intermingled with bundles of unlabelled axons and glial or ependymal processes. In the neurohemal contact zone, immunoreactive endings were observed among unlabelled neurosecretory endings in close vicinity to fenestrated capillary perivascular space. Small moderately intense immunofluorescent varicosities were observed all over the hypothalamus. The density of the glutamate decarboxylase-positive network was higher than in most diencephalic regions. Intraventricular or topical injection of colchicine allowed the visualization of small lightly immunoreactive cells in the diffusion area of colchicine. In the arcuate nucleus labelled axonal endings containing small pleomorphic synaptic vesicles and sometimes a few dense-core vesicles were observed at the electron-microscopic level. Typical synaptic junctions were commonly found between positive endings and unlabelled perikarya, or more frequently, unlabelled dendrites. These findings show that glutamate decarboxylase-containing endings are localized in several strategic sites for potential GABAergic neuroendocrine regulations. The GABAergic endings found among neurosecretory endings in the neurohemal contact zone may provide the morphological support for the release of γ -aminobutyrate into the portal blood flow as an hypothalamic hypophysiotropic hormone. Alternatively, neurosecretory cells might be under GABAergic control expressed either at their terminal level within the median eminence or the cell body level within the parvicellular hypothalamic nuclei.

Lesion‐induced mossy fibers to the molecular layer of the rat fascia dentata: Identification of postsynaptic granule cells by the Golgi‐EM technique

The axons of the dentate granule cells, the hippocampal mossy fibers, sprout "backward" into the dentate molecular layer when this is heavily denervated. Using the combined Golgi-electron microscopy (EM) technique we now demonstrate that these aberrant supragranular mossy fibers at least in part terminate on granule cell dendrites. Sprouting of mossy fibers into the dentate molecular layer was induced in adult rats by simultaneous surgical removal of the commissural and entorhinal afferents to the fascia dentata. After at least 7 weeks survival, the presence of mossy fiber terminals in the inner part of the dentate molecular layer was demonstrated by light microscopy. In the electron microscope the mossy fiber terminals were identified by their unique structural characteristics, namely, the unusually large size of the terminals, the dense packing of clear synaptic vesicles with a few dense core vesicles intermingled, the presence of asymmetric synaptic contacts with spines and desmosome-like contacts with dendritic shafts, and the continuity with a thin unmyelinated preterminal axon. Golgi-stained granule cells were first identified in the light microscope, and then, after deimpregnation, the same cells were examined in the electron microscope. In ultrathin, serial sections lesion-induced mossy fiber terminals were found in synaptic contact with spines on proximal dendritic segments of such identified Golgi-impregnated granule cells. From this we conclude that the aberrant, supragranular mossy fibers can innervate dendrites of the parent cell group, the dentate granule cells. The results, moreover, provide an example of reactive synaptogenesis where both the sprouted afferents and its postsynaptic element have been identified.

The fine structure of laminae IV, V, and VI of the Macaque spinal cord.

The normal fine structure of the deep dorsal horn (laminae IV, V, and VI) of the lumbosacral spinal cords of adult Macaque monkeys was examined. Axonal profiles with rounded synaptic vesicles (R) constitute about one-fourth of the total synaptic population in lamina IV and gradually increase in number to comprise more than a third of the synaptic population in lamina VI. Conversely, profiles with flattened vesicles (F) make up two-thirds of the synaptic population in lamina IV and slightly more than half in lamina VI. Axons which form the central profile (C) of synaptic glomeruli are more common in laminae IV and V than in lamina VI, and constitute about 5% of the total synaptic population. Profiles with large granular vesicles (LGV) are relatively uncommon in all the deep laminae. The vast majority of synaptic contacts are axodendritic. Axoaxonal synapses are formed between F, and R or C profiles, the F profile being the presynaptic component. It is concluded that there are both light and electron microscopic morphological differences among the deep laminae, as well as between the deep laminae and the superficial laminae of the spinal cord. Therefore, it appears that Rexed's original schema for laminar organization of the dorsal horn in the cat is also applicable to that of the monkey. In view of the relative paucity of LGV profiles in the deep dorsal horn, it is suggested that synapses in these laminae which contain monoamines or peptides are more likely to be associated with clear synaptic vesicles than granular synaptic vesicles.

Ultrastructure of normal and degenerating glomerular terminals of dorsal root axons in the substantia gelatinosa of the rhesus monkey.

The fine structure of primary sensory terminals within glomerular complexes of lamina II of Rexed (substantia gelatinosa Rolandi) in the spinal cord was investigated in normal adult rhesus monkeys and in monkeys subjected to thoracic or lumbosacral dorsal root transection. Three types of "scalloped" primary sensory terminals were distinguished on the basis of their ultrastructural characteristics, size, and distribution of synaptic vesicle population: (1) dense sinusoid axon (DSA) terminals contain medium-sized (42--46 nm and 58--62 nm) and large (80 nm) clear synaptic vesicles; (2) large dense-core vesicles (LDCV) terminals are equipped with empty synaptic vesicles ranging from 30 to 106 nm, large, (80 nm) and very large, (100 nm) dense-core vesicles; and (3) regular synaptic vesicles (RSV) terminals contain a homogeneous population of 45--50 nm clear synaptic vesicles. Following transection of the dorsal roots, all three types of primary afferents degenerate and become engulfed and phagocytosed by glial cells. However, each type of terminal displays a different mode and tempo of degeneration as seen in monkeys sacrificed 36, 48, and 72 hours following rhizotomy. DSAs follow the osmiophilic degeneration pattern; LDCVs are characterized by a gradual increase in the number of "electron-dense bodies" and, less frequently, by a progressive osmiophilic process; RSVs exhibit signs of a filamentous degeneration, accompanied by clusters of synaptic vesicles. The three types of terminals are distributed in an overlapping but distinct pattern within the posterior horn. Thus DSAs are present in highest numbers in lamina II where they constitute the most frequent terminal type. LDCVs also occur in lamina II in its outer half but are most concentrated in lamina I. RSVs predominate in the deeper layers of the dorsal horn (lamina III) but are also found in the internal half of lamina II. On the basis of these ultrastructural data and a comparison with afferent profiles impregnated according to the Golgi method, it appears that DSAs and LDCVs correspond respectively to superficial and marginal collaterals of small, thin dorsal root fibers whereas RSVs represent terminals of deep collaterals from large, thick dorsal root axons.

Synaptology of neurosecretory cells in the nucleus paraventricularis of the domestic fowl

Processes of magnocellular neurosecretory cells (MNCs) are easily identifiable on the basis of their content in neurosecretory granules in the neuropil of the rostral division of the paraventricular nucleus (PVN) of the domestic fowl. In specimens sacrificed during the winter the synaptic organization of the neuropil and the pattern of synapses ending on neurosecretory processes were studied at the ultrastructural level. Synapses in the rostral part of the PVN neuropil may be divided into three main categories on the basis of their morphology and their content of clear and dense-core synaptic vesicles. These different types of terminals can be attributed to aminergic, peptidergic or other types of synapses. The percent distribution of synapses within these categories differs when all synapses observed in the neuropil or only those ending on MNC processes are compared. Present ultrastructural data obtained in birds support two physiological hypotheses already suggested for mammals, i.e., the probable existence of a recurrent pathway to MNCs via an interneuron, and the importance of aminergic and peptidergic input in regulating the electrical activity ov MNCs.

Ultrastructure of hair follicle afferent fibre terminations in the spinal cord of the cat.

In acute electrophysiological experiments on anaesthetized cats, single identified hair follicle afferent fibres were injected with horseradish peroxidase (HRP). The HRP was injected from an intra-axonal microelectrode in the lumbosacral spinal cord. One to six hours after injection the animals were perfused and the tissue prepared for light and electron microscopy (EM). Axon collateral arborizations containing HRP reaction product were identified in thick sections under the light microscope and the same tissue then cut on the ultramicrotome for EM study. The terminal branches of the collaterals kept their myelin sheaths until they were 0.45-1.0 micron in diameter, just before they formed synaptic boutons. Synaptic boutons (1.0-4.0 microns in diameter) were usually of the en passant variety and made contact with dendrites. The contacts were asymmetrical (Type I) and contained round, clear synaptic vesicles of 35-60 nm diameter. Both the non-myelinated portion of the terminal axon and the synaptic boutons received axo-axonic contacts. These axo-axonic boutons contained clear (agranular) vesicles irregular in profile.

Light- and electron-microscopic characterization of electrophysiologically-identified, horseradish peroxidase-injected magnocellular neuroendocrine cells in goldfish preoptic nucleus

We recorded intracellularly from neurons in the goldfish preoptic nucleus which were anti dromically identified by electrical stimulation of the pituitary gland and marked by intracellular injection of horseradish peroxidase for subsequent localization. At the light-microscopic level, labeled neurons resembled profiles of Golgi-impregnated neurons and lay in the magnocellular portion of the preoptic nucleus. Densely labeled axons and dendrites projected to the lateral forebrain bundle, the medial forebrain bundle, fiber tracts in the preoptico-hypophysial tract, small blood vessels and capillaries, the ependymal lining of the third ventricle and toward other preoptic neurons. Occasionally, a lightly-labeled, large perikaryon lay adjacent to a large, heavily-labeled magnocellular neuron. Ultra structural examination of these identified cells revealed dense reaction product in neuronal perikarya and processes. Heavily labeled perikarya had elaborate networks of endoplasmic reticulum, extensive Golgi apparatus, occasional somatic spines and infrequent axo-somatic contacts from unlabeled neurons. These labeled perikarya which were frequently in close somatic apposition with unlabeled profiles were sometimes adjacent to a large, lightly-labeled perikaryon. A thin glial sheath separated most labeled neurons and processes from brain capillary endothelium. Labeled dendrites had heavily labeled spines and axo-dendritic contacts from unlabeled neurons. Labeled axons abutted unlabeled axons and -dendrites. Synaptic boutons innervating labeled structures always contained small clear synaptic vesicles and some boutons also contained large dense-core vesicles. These results demonstrate the complex connections of goldfish preoptic magnocellular neuroendocrine cells with other neurons, fiber systems, brain capillaries, ventricular ependyma and the pituitary and provide further support for non-endocrine as well as endocrine functions of magnocellular neurons.