Fine structure of nervous elements in the pancreas of some vertebrates

Summary The innervations of the exocrine and endocrine pancreas of some vertebrate animals were studied by electron microscopy. The pancreas of the bat and monkey contained ganglion cells in the interlobular connective tissue or between acinar cells. Unmyelinated nerve fibers ran through the interlobular connective tissue and reached the exocrine and endocrine parts, and terminated there as the endings. The nerve endings within the pancreas could be divided into four types: 1. Type 1-a of the nerve ending contained only agranular synaptic vesicles of about 500 Å in diameter. 2. Type 1-b characterized by containing agranular synaptic vesicles and some large cored vesicles (1,000 Å diameter). These two types of nerve endings might belong to the cholinergic (parasympathetic) endings. 3. Type 2-a contained small cored vesicles and agranular synaptic vesicles along with a few large cored vesicles. 4. Type 2-b was characterized by containing vesicles of the same size as those of agranular synaptic vesicles, and a majority of these vesicles contained bar-shaped crystalloids. This ending also contained a few large cored vesicles. These nerve endings of Type 2-a and 2-b might be the adrenergic (sympathetic) endings.     

A survey of the cytology and synaptic organization of the insular trigeminal-cuneatus lateralis nucleus in the rat, including an identification of spinal afferent inputs

The cytology and synaptic organization of the insular trigeminal-cuneatus lateralis (iV-Cul) nucleus was examined in the rat. In addition, the ultrastructural morphology and synaptic connectivity of anterogradely labeled spinal afferent axons terminating in iV-Cul were examined following injection of horseradish peroxidase (HRP) into the cervical spinal cord. The uniformity of the ultrastructural features of iV-Cul neurons supports the presence of a homogeneous neuronal population. The most prominent feature of the iV-Cul neuropil is the presence of numerous interdigitating astrocytic processes, which extensively isolate neuronal somata and processes. iV-Cul contains a heterogeneous population of axonal endings that can be separated into three categories, depending upon whether they contain predominantly spherical-shaped agranular synaptic vesicles (R endings), predominantly pleomorphic-shaped agranular synaptic vesicles (P endings), or a heterogeneous population of dense-core vesicles (DC endings). The R endings represent the majority of axonal endings in iV-Cul and establish asymmetrical axodendritic and axospinous synaptic contacts, primarily along the distal portions of the dendritic tree. P endings establish symmetrical axosomatic, axodendritic, and axospinous synaptic contacts and exhibit a more generalized distribution along the somadendritic tree. DC terminals establish asymmetrical axodendritic synaptic contacts with distal dendritic processes and are the least frequently observed endings in the iV-Cul neuropil. Numerous synaptic glomeruli, exhibiting a single large central R bouton that establishes multiple axodendritic or axospinous synapses, characterize the iV-Cul neuropil. Axoaxonic synapses are conspicuously absent from the iV-Cul neuropil and glomeruli. The anterograde HRP labeling of spinal afferent axons that terminate in iV-Cul indicates that the terminals along these fibers are of the R type and that they are engaged predominantly in synaptic glomeruli. The results of this study indicate that the synaptic organization of iV-Cul is distinctly different from that of neighboring somatosensory nuclei, and supports the recent suggestion that this nucleus should be considered a separate precerebellar spinal relay nucleus in the lateral medulla.     

AN ELECTRON MICROSCOPIC CHARACTERIZATION OF CLASSES OF SYNAPTIC VESICLES BY MEANS OF CONTROLLED ALDEHYDE FIXATION

Examination of variables of aldehyde fixation that may affect the shape of agranular synaptic vesicles has revealed that even brief storage of aldehyde-perfused nervous tissue pieces in cacodylate buffer, prior to hardening in osmium tetroxide, has an unusually severe flattening effect on agranular vesicles of a particular type. These are the vesicles of peripheral cholinergic axon endings, and of certain central synaptic bulbs. Types of synaptic bulbs can now be further defined on the basis of shape of agranular synaptic vesicles under controlled conditions of aldehyde fixation. Previously described "S" bulbs in the spinal cord contain uniformly spheroid vesicles, which are wholly resistant to flattening. Previously described "F" bulbs contain somewhat smaller agranular vesicles that are flattened after aldehyde fixation, even when this is followed by prompt hardening in osmium tetroxide solution. A third type, previously characterized as having irregularly round agranular vesicles after the above treatment, contains only severely flattened vesicles when the osmium tetroxide hardening is preceded by even a brief wash with sodium cacodylate buffer containing sucrose. Moreover, the "third" type is characteristic of all cholinergic peripheral axon endings examined, as well as the large axosomatic ("L") synaptic bulbs of the spinal cord.     

A Survey of the Cytology and Synaptic Organization of the Insular Trigeminal—Cuneatus Lateralis Nucleus in the Rat,Including an Identification of Spinal Afferent Inputs

The cytology and synaptic organization of the insular trigeminal—cuneatus lateralis (iV-Cul) nucleus was examined in the rat. In addition, the ultrastructural morphology and synaptic connectivity of anterogradely labeled spinal afferent axons terminating in iV-Cul were examined following injection of horseradish peroxidase (HRP) into the cervical spinal cord. The uniformity of the ultrastructural features of iV-Cul neurons supports the presence of a homogeneous neuronal population. The most prominent feature of the iV-Cul neuropil is the presence of numerous interdigitating astrocytic processes, which extensively isolate neuronal somata and processes. iV-Cul contains a heterogeneous population of axonal endings that can be separated into three categories, depending upon whether they contain predominantly spherical-shaped agranular synaptic vesicles (R endings), predominantly pleomorphic-shaped agranular synaptic vesicles (P endings), or a heterogeneous population of dense-core vesicles (DC endings). The R endings represent the majority of axonal endings in iV-Cul and establish asymmetrical axodendritic and axospinous synaptic contacts, primarily along the distal portions of the dendritic tree. P endings establish symmetrical axosomatic, axodendritic, and axospinous synaptic contacts and exhibit a more generalized distribution along the somadendritic tree. DC terminals establish asymmetrical axodendritic synaptic contacts with distal dendritic processes and are the least frequently observed endings in the iV-Cul neuropil. Numerous synaptic glomeruli, exhibiting a single large central R bouton that establishes multiple axodendritic or axospinous synapses, characterize the iV-Cul neuropil. Axoaxonic synapses are conspicuously absent from the iV-Cul neuropil and glomeruli. The anterograde HRP labeling of spinal afferent axons that terminate in iV-Cul indicates that the terminals along these fibers are of the R type and that they are engaged predominantly in synaptic glomeruli. The results of this study indicate that the synaptic organization of iV-Cul is distinctly different from that of neighboring somatosensory nuclei, and supports the recent suggestion that this nucleus should be considered a separate precerebellar spinal relay nucleus in the lateral medulla.     

Fine structural characteristics and synaptic connections of trigeminocerebellar projection neurons in rat trigeminal nucleus oralis

In order to classify the presynaptic terminals contacting trigeminocerebellar projection neurons (TCPNs) in rat trigeminal nucleus oralis (Vo), electron-microscopic examination of sequential thin sections made from TCPNs located in the border zone (BZ) of Vo, labeled by the retrograde transport of horseradish peroxidase, was undertaken. The use of BZ TCPNs, labeled in Golgi-like fashion so that many of their dendrites and axons were visible, allowed for the determination of the distribution of each bouton type along the soma and dendrites, as well as for the characterization of the morphology and synaptic relations of the labeled axon and its terminals. Three types of axon terminals contacting labeled BZ TCPNs have been recognized, depending upon whether they contain primarily spherical-shaped, agranular synaptic vesicles (S endings); predominantly flattened, agranular synaptic vesicles (F endings); or a population of pleomorphic-shaped, agranular synaptic vesicles (P endings). The S endings represent the majority of axon terminals contacting labeled BZ TCPNs and establish asymmetrical axosomatic and axodendritic synaptic contacts. Many S endings are situated in one of two types of synaptic glomeruli. One type of glomerulus has a large S ending at its core, whereas the other contains a small S ending. Large-S-ending glomeruli include only labeled distal dendrites of BZ TCPNs; small-S-ending glomeruli contain either a labeled soma, proximal dendrite, or distal dendritic shaft. The remaining S endings are extraglomerular, synapsing on distal dendrites. P endings are less frequently encountered and establish intermediate axosomatic and axodendritic synapses. These endings exhibit a generalized distribution along the entire somatodendritic tree. F endings make symmetrical axodendritic synapses with distal dendrites, are only found in glomeruli containing small S endings, and are the least frequently observed ending contacting labeled BZ TCPNs. The majority of axonal endings synapsing on labeled BZ TCPNs are located along distal dendrites, with only a relatively few synapsing terminals situated on proximal dendrites and somata. The axons of labeled BZ TCPNs arise from the cell body and generally give rise to a single short collateral near their points of origin. This collateral remains unbranched and generates several boutons within BZ, while the parent axon acquires a myelin sheath and, without branching further, travels dorsolaterally toward the inferior cerebellar peduncle. The collateral boutons resemble extraglomerular S endings. They contain agranular, spherical-shaped synaptic vesicles and make asymmetrical axodendritic synapses with small-diameter unlabeled dendritic shafts in the BZ neuropil.     

Nervous system of the snailHelix aspersa

Summary The fine structure of neurosecretory nerves and endings associated with the sheath of the infraesophageal ganglion ofHelix aspersa is described. The sheath is a neurohemal organ whose vascularized stroma receives both monoaminergic and peptidergic endings. The latter occur along the surface of the nerves or scattered within the stroma. They include a complex population of vesicular profiles. The granular vesicles (1300–3000 Å in diameter) exhibit structural modifications that may be related to the intra-axonal release of their neurohormones. The agranular vesicles (500–2000 Å in diameter) occur in large numbers and lie mostly adjacent to the axon surface. Synaptoid specializations seem to represent active sites for the extracellular discharge of neurosecretory material. The monoaminergic endings so far studied lack synaptoid specializations and contain small granular (800–1300 Å in diameter) and agranular (700 Å in diameter) vesicles. Two kinds of non-neural cells are associated with the nerves: glial cells and melanocytes.Partly supported by Conicyt (Grant 105) and Comisión de Investigación Científica Universidad de Chile (Grant 48). The technical assistance of Mr. Arnold van Dun is gratefully acknowledged. We also thank the Department of Physics, Faculty of Physical Sciences and Mathematics, University of Chile, for the use of a Philips EM-300 electron microscope.     

A comparative morphometric study of synaptic vesicles and other organelles in mossy fiber endings of the pigeon and the rat

The mossy fiber ending organelles of 4 rats and 4 pigeons were studied using ultrastructural morphometric methods. The number of agranular vesicles per square micrometer of synaptic surface, as well as the number of agranular vesicles per synapse, were found to be greater in the rat than in the pigeon, while no significant differences were found in coated and dense core vesicles. A highly positive correlation was found, in both species, between the synaptic surface and the number of agranular vesicles per unit volume of mossy fiber endings, while no correlation was found between the synaptic surface and numerical densities of coated and dense core vesicles.     

The zinc iodide-osmium tetroxide (ZIO) reaction on nerve endings in the median eminence of the rat under normal and experimental conditions

Summary The reaction of nerve endings in the median eminence of the rat to zinc iodide-osmium tetroxide (ZIO) staining was examined electron microscopically under normal and experimental conditions. The experimental condition of catecholamine exhaustion in the nerve endings was induced by the administration of H44/68 and reserpine. Vesicles in the terminals of catecholaminergic nerves reacted similarly to ZIO staining in both normal and experimental material. The majority of synaptic vesicles in various terminals gave a positive ZIO reaction. The neurosecretory elementary granules, however, failed to react with ZIO. On the other hand, some nerve terminals in the external layer of the median eminence showed a strong positive reaction in the cytoplasmic matrix, in mitochondria as well as in synaptic vesicles. These findings strongly suggest that the ZIO-positive substance in nerve terminals is not the transmitter itself, i.e. the monoamine, but rather represents a range of substances commonly found in various kinds of synaptic vesicles and is probably proteinaceous in nature. A brief discussion is also given on the difference in ZIO reactivity between neurosecretory elementary granules and small vesicles in the hypothalamo-hypophyseal tract.This work was supported in part by a research grant from the Ministry of Education, Japan     

Modifications in the neurosecretory system of the fish,Misgurnus fossilis,induced by lasting inanition

Summary The reaction of the hypothalamic neurosecretory system to prolonged starvation was examined in the fish,Misgurnus fossilis L. Two and four months of starvation resulted in storage of neurosecretory substance in the perikarya of the nucleus preopticus and in the preoptico-hypophysial tract. Further starvation, prolonged to five or seven months, caused degenerative changes in the nucleus preopticus. However, there were no drastic changes in the amount of stainable substance in the neurohypophysis. The examination of control animals revealed that the nucleus preopticus undergoes seasonal changes which differ from those brought about by starvation.     

Fine structure of islet-cell innervation in the pancreas of normal and alloxan-treated rats

Summary The innervation of the islets of Langerhans of normal albino rats and of albino rats treated with several daily doses of 125 mg/kg of alloxan was studied by electron microscopy. In the normal rat, nerve endings containing either agranular vesicles (200–400 Å) alone or in combination with large granular vesicles (500–800 Å) were found on both alpha and beta cells. Infrequently a third type of nerve ending containing small granular synaptic vesicles could be observed. Bundles of unmyelinated axons were also seen, as were typical autonomic ganglion cells. Similar normal neural elements were noted in rats treated with alloxan. However, islets of alloxan-treated animals also possess large elliptical profiles which appear to be dystrophic nerve terminals. These structures most frequently contact degranulated beta cells. Islets of Langerhans fixed with zinc iodide-osmium (ZIO) reported to specifically impregnate synaptic vesicles were also studied. Synaptic vesicles of normal axons and nerve endings as well as of the dystrophic structures were filled with ZIO reactive material. These studies suggest that alloxan may induce autonomic nerve ending changes in the rat endocrine pancreas. This may result from neuronal hyperactivity in an attempt to secrete insulin from the post-alloxan insulin-depleted beta cell.     

Ultrastructure of the hypothalamic neurosecretory system of the dog

Summary The hypothalamic neurosecretory system of normal dogs was studied by light and electron microscopy after perfusion-fixation. In the supraoptic nucleus most neurons are loaded with elementary neurosecretory granules having a content of low electron density. Neurons with less neurosecretory material and signs of enhanced synthetic activity, as recognized by the changes in the endoplasmic reticulum, were also observed.The vesiculated neurons ofJewell were studied under the electron microscope and various stages of development were described. It was postulated that they originate by a localized process of cytoplasmic cytolysis which ends in the formation of a large aqueous intracellular cavity limited by a plasma membrane. The possible significance of these vesiculated neurones is discussed. Some few myelinated neurosecretory axons are found in the supraoptic nucleus.The neurons of the paraventricular nucleus are smaller and contain less neurosecretory material. This is abundant and very pale in the axons. The median eminence consists of an inner zone, mainly occupied by the neurosecretory axons of the hypothalamic-neurohypophysial tracts, and an outer zone in which some neurosecretory axons end on the capillary of the portal system. This outer zone contains numerous axons with the axoplasm rich in neurofilaments and some containing granulated and non-granulated synaptic vesicles. Some neurons with granulated vesicles were observed in this region. The adrenergic nature of these neurons and axons is postulated.The infundibular process of the neurohypophysis shows small axons with discrete amounts of elementary granules and vesicles of synaptic type at the endings. Some enlarged axons having, in addition, large polymorphic bodies are observed and related to the Herring bodies.The size and morphology of the granules are analyzed along the entire hypothalamic-neurohypophysial system. The changes in diameter and electron density are related to the maturation of the granules and the possible significance of such evolution.Supported by grants from the Consejo Nacional de Investigaciones Cientificas y Técnicas and by the Air Force Office of Scientific Research (AF-AFOSR 963-66).     

Cerebrospinal fluid-contacting neurons,ciliated perikarya and “peptidergic” synapses in the magnocellular preoptic Nucleus of teleostean fishes

Summary The magnocellular preoptic nucleus of fishes (Anguilla anguilla, Amiurus nebulosus, Cyprinus carpio, Carassius auratus, Ctenopharyngodon idella, Cichlasoma nigrofasciatum) has been studied by light and electron microscopy.Two kinds of neurons were found: a) large, electron-dense, Gomori-positive cells with moderate acetylcholinesterase (AChE) positivity which contain granulated vesicles of 1400 to 2200 Å (in average 1600 to 1800 Å), and b) small, strongly AChE-positive, electron-lucent neurons containing granulated vesicles of 900 to 1200 Å. The nerve cells are supplied with axo-somatic and axo-dendritic synapses. These are formed by axon terminals containing either 1. synaptic vesicles of 500 Å, or 2. synaptic vesicles of 500 Å and dense-core vesicles of 600 to 800 Å, or 3. synaptic vesicles of 600 Å and granulated vesicles of up to 1100 Å, or 4. synaptic vesicles of about 400 Å and granulated vesicles of up to 1800 Å. The presence of peptidergic and numerous other synapses shows the complexity of the organization and afferentation of the magnocellular preoptic nucleus.In the eel, both types of nerve cells form dendritic terminals within the cerebrospinal fluid (CSF). These CSF contacting dendrites are supplied with 9×2+0 cilia. In the other species investigated, only some large neurons build up intraventricular endings. The ependymofugal process of the CSF contacting neurons enters the preoptic-neurohypophysial tract.Perikarya of both the large and the small cells may give rise to single, paired or multiple 9×2+0 cilia extending into the intercellular space. The number of CSF contacting neurons is reciprocal to the number of perikarya with intercellular cilium. These latter cells may represent modified, more differentiated forms of the CSF contacting neurons. We think that atypical cilia protruding into the intercellular space may have the same significance for the intercellular fluid as the cilia of the intraventricular dendrites of the CSF contacting neurons for the CSF.Dedicated to Prof. Dr. W. Bargmann on the occasion of his 70th birthday.     

Fine structure of the median eminence and the pars nervosa of the bullfrog,Rana catesbeiana

Summary The hypothalamic neurosecretory system of the bullfrog, Rana catesbeiana, was studied with light- and electron microscopy. The median eminence is roughly divided into two portions. The upper portion mostly consists of ependymal cells, glial cells and preoptico-hypophysial nerve tract, whereas in the lower portion, neurosecretory axons, glial cells, processes of glial and ependymal cells, and fine blood vessels of the hypothalamic portal vein are located. A part of the neurosecretory axons of the preoptico-hypophysial tract proceeds to the lower portion of the median eminence. These axons are arranged perpendicularly to the capillaries of the hypothalamic portal vein. The glial cells are densely located in the area of the median eminence where neurosecretory material is abundant. The neurosecretory material in the neurosecretory cells, their axons, the median eminence and the pars nervosa of the bullfrog shows a positive reaction to PAS treatment.The neurohemal area of the median eminence is occupied by many neurosecretory and non-neurosecretory axons, containing neurosecretory granules and/or synaptic vesicles. The axonal portions with the synaptic vesicles which are considered to be the nerve endings abut on the capillaries of the portal system. The size of synaptic vesicles in the axon terminals containing few neurosecretory granules is larger than those in the endings with many neurosecretory granules. Infrequently glial and ependymal processes are interposed between the nerve endings and the capillary wall.In the hilar region of the infundibulum, synapses are frequently observed between the thin fibers with or without neurosecretory granules and dendrites of non-neurosecretory neurons. The probable functions of these synapses are briefly discussed on the basis of our findings. Both in the hilar region of the infundibulum and in the pars nervosa, electron-dense neurosecretory granules of two different sizes were observed. The median eminence contains only one type of granules.The fine structure of the pars nervosa shows similar structures to those of the median eminence. Both in the median eminence and the pars nervosa, the fenestrated endothelium of the capillaries was frequently observed. The thick perivascular connective tissue space containing fibroblasts and collagen fibrils was observed both in the median eminence and the pars nervosa. Vesicles in the cytoplasm of the endothelial cells which appear to take a part in the transendothelial transport were observed.This investigation was supported in part by United States Public Health Service Research Grant, No. A-3678, to Hideshi Kobayashi from the National Institute of Arthritis and Metabolic Diseases and partly by a grant for Fundamental Scientific Research from the Ministry of Education of Japan. The authors wish to express their thanks to Prof. K. Takewaki for his kind encouragement.     

Ultrastructure of synapses in the nucleus of the trapezoid body of the bat]

The subsynaptic structure of the synapses in the medial nucleus of the trapezoid body was studied in the bat Myotis oxygnatus. The synaptic endings in the nucleus are represented by large-cup-shaped and small loop-shaped terminations. The cup-shaped terminations are formed of large branches originating from a thick myelinated fibre after loss of myelin from it. Each branch forms a series of contacts alternating with vast enlargements of extracellular space, on the body of the cell and its processes. Large branches are filled with synaptic vesicles, neurofilaments and neurotubules, mitochondria; all these components are distributed rather regularly along the branch diameter. In fine branches of the cup the synaptic vesicles are the main and often the only component. The pattern of the cup branch changes as the distance from the main fibre increases, namely the amount of neurofilaments and neurotubules diminishes up to their disappearance, while the amount and the density of synaptic vesicles increases. The small loop-shaped treminals are different from the cup-shaped ones by the composition of the synaptic vesicles and the structure of the contact zone. In addition to agranular vesicles there are also granular ones. Both types of terminations--cup-shaped and loop-shaped ones -- are found both on the bodies and dendrites. On distal portions of dendrites the terminations are disposed in nests.     

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.     

The hypothalamo-neurohypophysial system in Indian fresh-water goby, Glossogobius giuris (Ham.)

The hypothalamo-neurohypophysial neurosecretory system in Indian fresh-water goby, Glossogobius giuris (Ham.) has been described. The tractus preoptico-hypophyseus serves the function of a morphological and physiological connection between the hypothalamus and pituitary gland. In addition to main mass of the nucleus preopticus cells (cystine/cysteine bearing), a group of few cells in the hypothalamus has also been observed. These cells are situated posterior to the position of the nucleus preopricus and are CH Ph + ve and AF + ve. The neurosecretory material in the cells of nucleus preopticus is in the form of fine granules. The nucleus lateralis tuberis is absent in the fish under study. The disposition of neurosecretory material is heaviest along the fibres of the neurohypophysis in the region of pars intermedia with which it forms a profuse interdigitation. The fibres usually terminate over the blood vessels. The Herring bodies are noticeable at different levels in the neurohypophysis and pars-distalis. Besides the neurosecretory fibres, Herring bodies, non-stainableneurosecretory fibres and blood vessels, the pituicytes are also present in the neurohypophysis (SAKSENA 1974a, b). The intraaxonal flow of neurosecretory material, the vascularization of the nucleus preopticus and hypothalamo-hypophysial regulatory mechanism have been also discussed.     

Hypothalamo-hypophysial Neurosecretory System in Normal and Hypophysectomised Catfish,Clarias Batrachus L

Two neurosecretory centers, nucleus preopticus (NPO) and nucleus lateralis tuberis (NLT), are distinguished in the hypothalamus of Clarias batrachus L. The cell bodies of NPO are grouped into nucleus preopticus magnocellularis (NPM) and nucleus preopticus parvocellularis (NPP). The NLT is recognised into three divisions: pars rostralis, pars medialis and pars ventrolateralis. The neurons of both the NLT and NPO are aldehyde-fuchsin (AF) positive. The axons from NPO run into four or five bundles ventrolaterally and caudally for some distance before all of them from either side join ventromedially into a single thick cord of neurosecretory fibers which finally enters the pars intermedia to form the neurointermediate lobe. The fibers from NLT, which are separate from the neurosecretory fibers of the NPO, anastomose in the region of the pars distalis. The neurosecretory material (NSM) of NPO is transported by axonal route whereas that of NLT is by axonal as well as ependymovascular pathways. Hypophysectomy results in the increase of AF-positive material in the neurons of NPO shortly after the operation, but later on, the material begins to deplete in them. The AF-positive material at the cut end of neurosecretory fibers, however, accumulates. The AF-positive material in the cell bodies of NLT is also depleted, but the nuclei increase in size, after hypophysectomy.     

Special dendritic and axonal endings formed by the cerebrospinal fluid contacting neurons of the spinal cord

Summary The cerebrospinal fluid (CSF) contacting neurons have a dendritic process which protrudes into the central canal, and is provided with one long kinocilium and many shorter stereocilia (about 80 in the turtle) as revealed by scanning electron microscopy. The shape, number and arrangement of the cilia are similar to those of known receptor endings.The silver impregnated axons of these cells converge to a paired centrosuperficial tract forming terminal enlargements at the ventrolateral surface of the spinal cord. Lying among glial endfeet these terminals are ultrastructurally similar to those present in known neurosecretory areas. The nerve endings are attached to the basal lamina, and they comprise many synaptic vesicles (200 to 400 Å in diameter), as well as granular vesicles of different sizes (diameter 600 to 1800 Å). The axons may lie within finger-like protrusions on the surface of the spinal cord, or they may terminate around vessels.Morphological evidence suggests that these nerve terminals and the corresponding CSF contacting perikarya represent a spinal neurosecretory system possibly influenced by information taken up by its special dendrites protruding into the inner CSF space.     

FINE STRUCTURE OF THE MEDIAL NUCLEUS OF THE TRAPEZOID BODY OF THE BAT WITH SPECIAL REFERENCE TO TWO TYPES OF SYNAPTIC ENDINGS

The medial nucleus of the trapezoid body has been studied electron microscopically in two species of bat, Miniopterus schreibersi fuliginosus and Vespertilio superans, which were perfused with three different kinds of fixatives, osmium tetroxide, glutaraldehyde, and formaldehyde. Two types of synaptic endings are observed in the nucleus: the abundant calyciferous endings and the less frequently occurring "small-vesicle endings." The former endings vary greatly in size, and contain extended extracellular spaces between pre- and post-synaptic membranes. The latter endings are always small, without the extended extracellular spaces, and tend to lie side by side. In all of the materials perfused with three different fixatives, synaptic vesicles in the calyciferous endings are round in shape and larger than those in the small-vesicle endings. The shape of vesicles in the small-vesicle endings varies according to the kinds of fixatives used; round in osmium tetroxide-fixed materials, flattened in formaldehyde-fixed materials, and somewhat round or flattened in glutaraldehyde-fixed materials. It is suggested that the calyciferous endings are excitatory in nature and that the small-vesicle endings are inhibitory.     

Microvesicles of the neurohypophysis are biochemically related to small synaptic vesicles of presynaptic nerve terminals

Nerve endings of the posterior pituitary are densely populated by dense- core neurosecretory granules which are the storage sites for peptide neurohormones. In addition, they contain numerous clear microvesicles which are the same size as small synaptic vesicles of typical presynaptic nerve terminals. Several of the major proteins of small synaptic vesicles of presynaptic nerve terminals are present at high concentration in the posterior pituitary. We have now investigated the subcellular localization of such proteins. By immunogold electron microscopy carried out on bovine neurohypophysis we have found that three of these proteins, synapsin I, Protein III, and synaptophysin (protein p38) were concentrated on microvesicles but were not detectable in the membranes of neurosecretory granules. In addition, we have studied the distribution of the same proteins and of the synaptic vesicle protein p65 in subcellular fractions of bovine posterior pituitaries obtained by sucrose density centrifugation. We have found that the intrinsic membrane proteins synaptophysin and p65 had an identical distribution and were restricted to low density fractions of the gradient which contained numerous clear microvesicles with a size range the same as that of small synaptic vesicles. The peripheral membrane proteins synapsin I and Protein III exhibited a broader distribution extending into the denser part of the gradient. However, the amount of these proteins clearly declined in the fractions preceding the peak of neurosecretory granules. Our results suggest that microvesicles of the neurohypophysis are biochemically related to small synaptic vesicles of all other nerve terminals and argue against the hypothesis that such vesicles represent an endocytic byproduct of exocytosis of neurosecretory granules.