Pleiomorphic Synaptic Vesicles Research Articles

Synaptic circuitry in the retinorecipient layers of the optic tectum of the lamprey (Lampetra fluviatilis). A combined hodological, GABA and glutamate immunocytochemical study

The ultrastructure of the retinorecipient layers of the lamprey optic tectum was analysed using tract tracing techniques combined with GABA and glutamate immunocytochemistry. Two types of neurons were identified; a population of large GABA-immunonegative cells, and a population of smaller, highly GABA-immunoreactive interneurons, some of whose dendrites contain synaptic vesicles (DCSV). Five types of axon terminals were identified and divided into two major categories. The first of these are GABA-immunonegative, highly glutamate-immunoreactive, contain round synaptic vesicles, make asymmetrical synaptic contacts, and can in turn be divided into AT1 and AT2 terminals. The AT1 terminals are those of the retinotectal projection. The origin of the nonretinal AT2 terminals could not be determined. AT1 and AT2 terminals establish synaptic contacts with DCSV, with dendrites of the retinopetal neurons (DRN), and with conventional dendritic (D) profiles. The terminals of the second category are GABA-immunoreactive and can similarly be divided into AT3 and AT4 terminals. The AT3 terminals contain pleiomorphic synaptic vesicles and make symmetrical synaptic contacts for the most part with glutamate-immunoreactive D profiles. The AT4 terminals contain rounded synaptic vesicles and make asymmetrical synaptic contacts with DRN, with DCSV, and with D profiles. A fifth, rarely observed category of terminals (AT5) contain both clear synaptic vesicles and a large number of dense-core vesicles. Synaptic triads involving AT1, AT2 or AT4 terminals are rare. Our findings are compared to these of previous studies of the fine structure and immunochemical properties of the retinorecipient layers of the optic tectum or superior colliculus of Gnathostomes.

Fine structure of the visual dorsolateral anterior thalamic nucleus of the pigeon (Columba livia): A hodological and GABA‐immunocytochemical study

The ultrastructure of the lateroventral subcomponent of the visual dorsolateral anterior thalamic nucleus of the pigeon (DLLv) was analyzed using hodological techniques and GABA-immunocytochemistry. Two types of GABA-immunonegative hyperpalliopetal neurons and a single type of strongly GABA-immunoreactive (-ir) interneuron were identified, the latter displaying long dendrites with some containing synaptic vesicles (DCSV). Ten types of axon terminal were identified and divided into two categories. The first, GABA-immunonegative and making asymmetrical synaptic contact, contain round (RT1, RT2, RT3) or pleiomorphic synaptic and many dense-core vesicles (DCT). RT1 terminals are retinothalamic and RT2 terminals hyperpalliothalamic; both mainly contact dendrites of projection neurons (72% and 78% respectively), less frequently dendrites of interneurons and sometimes DCSV; RT1 terminals are rarely involved in synaptic triads. The second category are consistently GABA-immunopositive. Four types (PT1-4), distinguished by their pleiomorphic synaptic vesicles, make symmetrical synaptic contact essentially with dendrites of projection neurons, more rarely on dendrites of interneurons (PT2). PT1 terminals are very probably those of interneurons, whereas the rare PT4 terminals are of retinal origin. A fifth type (RgT) contains round synaptic vesicles and makes asymmetrical synaptic contact with dendrites of projection neurons and interneurons. PT2 and RgT terminals occasionally contact DCSV of interneurons, which are sometimes involved in synaptic triads. Two final subcategories (DCgT1-2) contain many dense-core vesicles. Our findings are compared with those of previous studies concerning the fine structure and neurochemical properties of the GLd of reptiles and mammals, with special reference to the origin of the extraretinal and extracortical projections to this structure.

Vestibuloocular reflex of the adult flatfish. III. A species-specific reciprocal pattern of excitation and inhibition.

In juvenile flatfish the vestibuloocular reflex (VOR) circuitry that underlies compensatory eye movements adapts to a 90 degrees relative displacement of vestibular and oculomotor reference frames during metamorphosis. VOR pathways are rearranged to allow horizontal canal-activated second-order vestibular neurons in adult flatfish to control extraocular motoneurons innervating vertical eye muscles. This study describes the anatomy and physiology of identified flatfish-specific excitatory and inhibitory vestibular pathways. In antidromically identified oculomotor and trochlear motoneurons, excitatory postsynaptic potentials (EPSPs) were elicited after electrical stimulation of the horizontal canal nerve expected to provide excitatory input. Electrotonic depolarizations (0.8-0.9 ms) preceded small amplitude (<0.5 mV) chemical EPSPs at 1.2-1.6 ms with much larger EPSPs (>1 mV) recorded around 2.5 ms. Stimulation of the opposite horizontal canal nerve produced inhibitory postsynaptic potentials (IPSPs) at a disynaptic latency of 1.6-1.8 ms that were depolarizing at membrane resting potentials around -60 mV. Injection of chloride ions increased IPSP amplitude, and current-clamp analysis showed the IPSP equilibrium potential to be near the membrane resting potential. Repeated electrical stimulation of either the excitatory or inhibitory horizontal canal vestibular nerve greatly increased the amplitude of the respective synaptic responses. These observations suggest that the large terminal arborizations of each VOR neuron imposes an electrotonic load requiring multiple action potentials to maximize synaptic efficacy. GABA antibodies labeled axons in the medial longitudinal fasciculus (MLF) some of which were hypothesized to originate from horizontal canal-activated inhibitory vestibular neurons. GABAergic terminal arborizations were distributed largely on the somata and proximal dendrites of oculomotor and trochlear motoneurons. These findings suggest that the species-specific horizontal canal inhibitory pathway exhibits similar electrophysiological and synaptic transmitter profiles as the anterior and posterior canal inhibitory projections to oculomotor and trochlear motoneurons. Electron microscopy showed axosomatic and axodendritic synaptic endings containing spheroidal synaptic vesicles to establish chemical excitatory synaptic contacts characterized by asymmetrical pre/postsynaptic membrane specializations as well as gap junctional contacts consistent with electrotonic coupling. Another type of axosomatic synaptic ending contained pleiomorphic synaptic vesicles forming chemical, presumed inhibitory, synaptic contacts on motoneurons that never included gap junctions. Altogether these data provide electrophysiological, immunohistochemical, and ultrastructural evidence for reciprocal excitatory/inhibitory organization of the novel vestibulooculomotor projections in adult flatfish. The appearance of unique second-order vestibular neurons linking the horizontal canal to vertical oculomotor neurons suggests that reciprocal excitation and inhibition are a fundamental, developmentally linked trait of compensatory eye movement circuits in vertebrates.

Types, numbers and distribution of synapses on the dendritic tree of an identified visual interneuron in the brain of the locust.

The descending contralateral movement detector (DCMD), an identified descending interneuron in the brain of the locust Schistocerca gregaria has been investigated by using light and electron microscopy. We describe the fine structure, distribution and numbers of synapses that it receives from another identified brain neuron, the lobular giant movement detector (LGMD), and from unidentified neurons. The DCMD dendrites emerging from the integrative segment vary in form and number between individuals and sexes but always form a flattened dendritic domain. The arborizations and the integrative segment appear to be exclusively postsynaptic. Two types of synaptic contacts (Type 1 and 2) onto the DCMD can be discerned as having either round (Type 1) or pleiomorphic synaptic vesicles (Type 2) and by large (Type 1) or small (Type 2) subsynaptic appositions. Contact zones of Type 1 synapses are smaller than those of Type 2. LGMD-synapses are of Type 1 and occur intermingled with presynaptic sites of unidentified units. Some branches of the DCMD receiving input from unidentified units are devoid of contacting LGMD processes. Synapses of both types are randomly distributed over the DCMD integrative segment and at fibres with similar sizes. Type 1 synapses are much more frequent than Type 2 synapses and their number is negatively correlated with fibre diameter. For a whole DCMD dendritic arborization, a total of 8500 active zones of chemical synapses has been calculated, including a minimum of 2250 LGMD-synapses and about 1000 Type 2 synapses. The DCMD may thus receive a considerable amount of input from as yet unidentified neurons.

Excitatory and Inhibitory Vestibular Pathways to the Extraocular Motor Nuclei in Goldfish

Electrophysiological, ultrastructural, and immunohistochemical techniques were utilized to describe the excitatory and inhibitory vestibular innervation of extraocular motor nuclei in the goldfish. In antidromically activated oculomotor motoneurons, electrical stimulation of the intact contralateral vestibular nerve produced short-latency, variable amplitude electrotonic excitatory postsynaptic potentials (EPSPs) at 0.5-0.7 ms followed by chemical EPSPs at 1.0-1.3 ms. Stimulation of the ipsilateral vestibular nerve produced small amplitude membrane hyperpolarizations at a latency of 1.3-1.7 ms in which equilibrium potentials were slightly more negative than resting potentials. The inhibitory postsynaptic potentials (IPSPs) reversed with large amplitudes after the injection of chloride ions suggesting a proximal soma-dendritic location of terminals exhibiting high efficacy inhibitory synaptic conductances. In antidromically identified abducens motoneurons and putative internuclear neurons, electrical stimulation of the contralateral vestibular nerve produced large-amplitude, short-latency electrotonic EPSPs at 0.5 ms followed by chemical depolarizations at 1.2-1.3 ms. Stimulation of the ipsilateral vestibular nerve evoked IPSPs at 1.4 ms that were reversed after injection of current and/or chloride ions. gamma-Aminobutyric acid (GABA) antibodies labeled inhibitory neurons in vestibular subdivisions with axons projecting into the ipsilateral medial longitudinal fasciculus (MLF). Putative GABAergic terminals surrounded oculomotor, but not abducens, motoneurons retrogradely labeled with horseradish peroxidase. Hence the spatial distribution of GABAergic neurons and terminals appears highly similar in the vestibuloocular system of goldfish and mammals. Electron microscopy of motoneurons in the oculomotor and abducens nucleus showed axosomatic and axodendritic synaptic endings containing spheroidal synaptic vesicles establishing chemical, presumed excitatory, synaptic contacts with asymmetric pre- and/or postsynaptic membrane specializations. The majority of contacts with spheroidal vesicles displayed gap junctions in which the chemical and electrotonic synapses were either en face to dissimilar or adjacent to one another on the same soma/dendritic profiles. Another separate set of axosomatic synaptic endings, presumed to be inhibitory, contained pleiomorphic synaptic vesicles with symmetric pre- and/or postsynaptic membrane specializations that never included gap junctions. Excitatory and inhibitory synaptic contacts appeared equal in number but were more sparsely distributed along the soma-dendritic profiles of oculomotor as compared with abducens motoneurons. Collectively these data provide evidence for both disynaptic vestibular inhibition and excitation in all subdivisions of the extraocular motor nuclei suggesting the basic vestibulooculomotor blueprint to be conserved among vertebrates. We propose that unique vestibular neurons, transmitters, pathways, and synaptic arborizations are homologous structural traits that have been essentially preserved throughout vertebrate phylogeny by a shared developmental plan.

Morphology and soma‐dendritic distribution of synaptic endings from the rostral interstitial nucleus of the medial longitudinal fasciculus (riMLF) on motoneurons in the oculomotor and trochlear nuclei in the cat

The morphology and soma-dendritic distribution of anterograde biocytin-labelled rostral interstitial nucleus of the medial longitudinal fasciculus (riMLF) synaptic endings in the oculomotor and trochlear nuclei have been examined by electron microscopy by using both preembedding immunoperoxidase and postembedding immunogold methods. The results indicate that three morphological types of riMLF synaptic endings are distinguishable on the basis of synaptic vesicle morphology (spheroidal, pleiomorphic, or ellipsoidal) and postsynaptic membrane specializations (asymmetrical or symmetrical). All three morphological types of riMLF synaptic endings establish synaptic connections predominantly with dendrites. Synaptic endings that contain ellipsoidal synaptic vesicles have a more proximal soma-dendritic distribution than those that contain either spheroidal or pleiomorphic synaptic vesicles. Furthermore, all three morphological types of synaptic endings are encountered in the same motoneuron subdivisions of the oculomotor and trochlear nuclei in the same experiments. The findings suggest that subregions of the riMLF contain coexistent populations of excitatory and inhibitory premotor neurons that are related to opposite directions of vertical saccadic eye movements but that project to the same motoneuron subgroups on the ipsilateral side. Both the morphology and the mode, pattern, and soma-dendritic distribution of saccade-related riMLF synaptic endings that establish synaptic connections with vertical motoneurons differ from those of excitatory and inhibitory second-order vertical vestibular synaptic endings. These differences in the synaptic organization of riMLF and second-order vestibular inputs to oculomotor and trochlear motoneurons may be related to differences in the information transferred by each source, the riMLF input conveying eye-velocity signals, and the vestibular input conveying eye-position signals. © 1996 Wiley-Liss, Inc.

Morphology and soma-dendritic distribution of synaptic endings from the rostral interstitial nucleus of the medial longitudinal fasciculus (riMLF) on motoneurons in the oculomotor and trochlear nuclei in the cat.

The morphology and soma-dendritic distribution of anterograde biocytin-labelled rostral interstitial nucleus of the medial longitudinal fasciculus (riMLF) synaptic endings in the oculomotor and trochlear nuclei have been examined by electron microscopy by using both preembedding immunoperoxidase and postembedding immunogold methods. The results indicate that three morphological types of riMLF synaptic endings are distinguishable on the basis of synaptic vesicle morphology (spheroidal, pleiomorphic, or ellipsoidal) and postsynaptic membrane specializations (asymmetrical or symmetrical). All three morphological types of riMLF synaptic endings establish synaptic connections predominantly with dendrites. Synaptic endings that contain ellipsoidal synaptic vesicles have a more proximal soma-dendritic distribution than those that contain either spheroidal or pleiomorphic synaptic vesicles. Furthermore, all three morphological types of synaptic endings are encountered in the same motoneuron subdivisions of the oculomotor and trochlear nuclei in the same experiments. The findings suggest that subregions of the riMLF contain coexistent populations of excitatory and inhibitory premotor neurons that are related to opposite directions of vertical saccadic eye movements but that project to the same motoneuron subgroups on the ipsilateral side. Both the morphology and the mode, pattern, and soma-dendritic distribution of saccade-related riMLF synaptic endings that establish synaptic connections with vertical motoneurons differ from those of excitatory and inhibitory second-order vertical vestibular synaptic endings. These differences in the synaptic organization of riMLF and second-order vestibular inputs to oculomotor and trochlear motoneurons may be related to differences in the information transferred by each source, the riMLF input conveying eye-velocity signals, and the vestibular input conveying eye-position signals.

Distribution of GABA immunoreactivity in the retino-recipient layer of the viper optic tectum. A light and electron microscope quantitative study.

The distribution of GABA-immunoreactivity was investigated in the principal retino-recipient layer of the optic tectum in the snake Vipera aspis. This layer, the stratum griseum et fibrosum superficiale, contained an important proportion (approximately 50%) of small GABA-immunoreactive interneurons, characterized by a voluminous invaginated nucleus surrounded by a thin rim of cytoplasm poor in organelles and occasionally showing pleiomorphic synaptic vesicles, which could also be observed in some of the dendrites that contained synaptic vesicles. In the neuropile, the GABA-immunoreactive profiles containing synaptic vesicles could be subdivided into dendrites containing synaptic vesicles and axon terminals with pleiomorphic synaptic vesicles. The dendrites containing synaptic vesicles (23.4% of all profiles containing synaptic vesicles) were postsynaptic either to optic terminals (39.2%), GABA-immunoreactive axon terminals with pleiomorphic synaptic vesicles (48.2%) or to immunonegative (S1) boutons with round synaptic vesicles (12.6%). These dendrites were presynaptic to GABA-immunoreactive (18%) neurons or immunonegative (82%) neurons. The axon terminals with pleiomorphic synaptic vesicles, which represented 47.4% of all profiles, were predominantly (99%) GABA-immunoreactive and four types could be distinguished according to cytological criteria. These axon terminals made synaptic contacts for the most part (78%) with immunonegative profiles, and more rarely (22%) with immunoreactive neurons. These data are compared to those previously obtained in the homologous structure of other vertebrate species, birds and mammals in particular.

Immunohistochemical localization of gaba in the mammillary complex of the rat

The distribution and synaptic organization of GABAergic elements in the mammillary nuclei of rats have been examined by the immunocytochemical localization of GABA at the light and electron microscope levels. The distribution of GABA-immunoreactive fibres and terminals in the mammillary body is non-homogeneous. By light microscopy, small scattered immunoreactive terminals are observed in the pars medianus, pars posterior and ventral region of the pars medialis of the medial mammillary nucleus. Larger labelled terminals are found in the pars lateralis, the dorsal region of the pars medialis of the medial mammillary nucleus and the lateral mammillary nucleus. At the ultrastructural level, GABA-immunoreactive synaptic endings in the different subdivisions of the medial mammillary nucleus exhibit a widespread somadendritic distribution. By contrast, GABA-immunoreactive terminals within the lateral mammillary nucleus are located predominantly in the neuropil and less frequently on neuronal somata. GABA-immunoreactive synaptic endings contain pleiomorphic synaptic vesicles and have symmetrical synaptic contact zones with the somata and dendrites in the lateral and medial mammillary nuclei. After in vivo inhibition of GABA metabolism with amino-oxyacetic acid, light microscopic examination of the mammillary nuclei reveals numerous small GABA-immunoreactive cells in various subdivisions of the medial mammillary nucleus. No immunoreactive cells are observed, however, in the lateral mammillary nucleus. Electron microscopic examination demonstrates that the GABA-immunoreactive cells are astrocytes. In the labelled astrocytes, immunohistochemical reaction product is localized throughout the nucleus and cytoplasm of the cells, in thin sheet-like processes surrounding neuronal elements and in end-feet lining the basal lamina of capillaries. The results indicate that the mammillary nuclei in the rat receive a strong GABAergic innervation. Most if not all, of the GABA-immunoreactive synaptic endings in the mammillary nuclei probably arise from extrinsic inhibitory sources. The possible sources of the GABA-immunoreactive input to the mammillary complex are discussed.

Presumptive GABAergic centrifugal input to the lamprey retina: a double-labeling study with axonal tracing and GABA immunocytochemistry

The distribution of GABA-like immunoreactivity (GABA-LI) was performed in the lamprey retinopetal system which was previously identified by either anterograde or retrograde axonal tracing methods. This study was carried out at the ultrastructural level for the retina and under both the light and electron microscope for the mesencephalic retinopetal centers (M5 and RMA). The GABA-LI was distributed in about 40% of anterogradely HRP-labeled axon terminals in the inner retina. These made synaptic contacts upon either HRP-labeled ganglion cell dendrites or mostly on GABA-LI or on immunonegative amacrine cell dendrites and somata. The other immunonegative HRP-labeled axon terminals also established synaptic contacts on amacrine cell dendrites and somata. The mesencephalic retinopetal neurons, retrogradely labeled with HRP or [ 3H]proline, were GABA-LI in 65% of M5 somata and only in 15% of RMA neurons. M5 and RMA retinopetal neurons and dendrites, either GABA-LI or immunonegative, were contacted: (1) asymmetrically by HRP-labeled or unlabeled axon terminals containing rounded synaptic vesicles, always immunonegative and (2) symmetrically by HRP-unlabeled axon terminals containing pleiomorphic synaptic vesicles, which were either GABA-LI or immunonegative. The role of GABA as a putative neurotransmitter in the centrifugal visual system is discussed.

Evidence of GABA-immunopositive neurons in the dorsal part of the lateral geniculate nucleus of reptiles: Morphological correlates with interneurons

The distribution and staining pattern of γ-aminobutyric acid immunoreactivity have been examined by both light and electron microscopy in the dorsal part of the lateral geniculate nucleus of three reptilian species: the turtle Chinemys reevesi, the lizard Ophisaurus apodus and the snake Vipera aspis. After perfusion of the animals with 1% paraformaldehyde and 1% glutaraldehyde and polyethyleneglycol embedding of the brains, the analysis of sections processed immunocytochemically with an anti-GABA antiserum has revealed a moderate-to-dense labeling of the neurons of the dorsal part of the lateral geniculate complex in these species. Labeled cell bodies are small-sized, either rounded or fusiform and the GABA-positive dendrites emerging from them are not preferentially oriented in any particular direction. Quantitative studies in Vipera indicate that GABA-positive neurons make up about 14% of the population of neurons of the dorsal part of the lateral geniculate nucleus. Electron microscopy of specimens treated by either pre- or post-embedding techniques has confirmed that these cells corresponded to neurons. No glial cells were ever observed to be immunopositive. These GABA-positive neurons, characterized by the presence of pleiomorphic synaptic vesicles localized either in their perikaryon or more often in presynaptic dendrites, established symmetrical synaptic contacts. In this case, the latter were involved both pre- and postsynaptically in serial and, more rarely, in triadic arrangements, a synaptic organization specific to interneurons. The involvement of such GABA-positive neurons in local circuits is discussed.

Fine structure of the optic fiber termination layer in the tectum of the teleost Rutilus: a stereological and morphometric study.

The stratum fibrosum et griseum superficiale (SFGS) of the Rutilus optic tectum, which receives a massive fiber projection from the contralateral retina, was studied by electron microscopy. The qualitative and quantitative analysis of the laterodorsal (LD) portion of the stratum involved both a stereological examination of the different elements and a morphometric study of the various profiles containing synaptic vesicles (PCSVs). The relative volume of each element in the LD SFGS was as follows: myelinated and unmyelinated axons, 6.6%; PCSVs, 38%; dendrites without vesicles, spines, and cell bodies, 41.7%; glia, 10.5%. With the fixation employed, 35% of PCSVs showed spheroidal synaptic vesicles. These profiles could be subdivided into three types: (1) S1 (23.5%) represented optic terminals, since they degenerated after retinal ablation or were labeled after intraocular injection of HRP or [3H] proline. Three subgroups of S1 were identified: S1m--profiles containing clear mitochondria;S1c--profiles that were contiguous with S1m and lacked mitochondria;S1i--isolated profiles without mitochondria. (2) S2 (9.3%) were characterized mainly by their dark mitochondria. (3) S3 (2.2%) corresponded to small nonvisual terminals that were isolated and lacked mitochondria. The PCSVs with pleiomorphic synaptic vesicles (65%) were subdivided into three groups: P1 (38%), P2 (19%), and P3 (8%). P1 and P2 were axonal in nature; P2 could be distinguished from P1 by a greater density of synaptic vesicles. P3 was of dendritic origin. Analysis of synaptic patterns revealed a small number of serial synapses. The presynaptic elements were optic boutons, whereas the intermediate profiles were dendrites with synaptic vesicles (P3). Results are compared with ultrastructural data obtained in the superficial tectal layers of other teleosts and other vertebrate groups.

Morphological studies of synapses and varicosities in dissociated cell cultures of sympathetic neurons.

Neurons dissociated from the superior cervical ganglia of newborn rats can be grown under conditions which support either adrenergic or cholinergic differentiation. In both cases, the neurons form numerous morphologically specialized synaptic terminals or synapses as well as relatively unspecialized varicosities. The ultrastructure of both types of terminal was compared in mature neuronal cultures and the effects of growth conditions on terminal morphology examined. After aldehyde-osmium fixation, synapses in cultures grown under adrenergic or cholinergic conditions were characterized by asymmetrical membrane specializations comparable to type I or asymmetric synapses; bismuth iodide and ethanolic phosphotungstic acid impregnation of neuronal cultures revealed the presence of characteristic synaptic membrane specializations: a presynaptic grid of dense projections and a wide postsynaptic dense band of uniform thickness. No membrane specializations were apparent in varicosities after aldehyde-osmium fixations or with these stains. Intramembranous particle distributions were examined in freeze-fracture replicas of neurons. Aggregates of large, 10-12 nm particles were found on P-face membrane leaflets of cell bodies and large diameter processes; this distribution is the same as that of synapses in thin-sectioned preparations. These particle aggregates may represent postsynaptic membrane specializations or acetylcholine receptors. The cytoplasmic leaflet of boutons contained large, 12-14 nm particles, which appeared to be concentrated at the region of synaptic contact at putative synapses, but were diffusely distributed in varicosity membranes. Similar large particles were also seen at a much lower density in the membrane E-face. None of these ultrastructural characteristics appeared to vary with transmitter identity or growth conditions. Synaptic vesicle shape, however, did vary in glutaraldehyde-fixed cultures. At all ages examined, neurons grown on monolayers of heart cells contained predominantly round vesicles, whereas neurons grown in the virtual absence of non-neuronal cells possessed pleiomorphic synaptic vesicles. This difference in vesicle shape appeared to be correlated more closely with growth in the presence of non-neuronal cells than with the transmitter present at the time of fixation.

Morphologically distinct classes of inhibitory synapses arise from the same neurons: Ultrastructural identification from crossed vestibular interneurons intracellularly stained with HRP

AbstractPrevious studies have demonstrated that in goldfish and tench inhibitory interneurons mediating both electrical and chemical inhibition of the Mauthner cell can be identified by a passive hyperpolarizing potential (PHP) which is recorded intracellularly when the latter is firing; some of them are crossed second‐order vestibular cells, the axons of which project ipsi‐laterally and contralaterally to their cell's body in several medullary nuclei. Since PHP‐exhibiting neurons and their target Mauthner cells can be simultaneously impaled, commissural interneurons were intracellularly stained with HRP contained in presynaptic microelectrodes and their synaptic features were correlated with electrophysiological findings. In this study we were able to confirm that commissural neurons are monosynaptically activated by VIIIth nerve afferents and that spikes selectively evoked in these cells produce monosynaptic Cl−‐dependent unitary IPSPs in their adjacent Mauthner cell; in addition to observations related to their general morphology, including their mode of termination within vestibular and reticular nuclei, new data were obtained, most of them with EM techniques. (1) The Mauthner cell is covered by well‐segregated synaptic endings of different shapes; among them, the unmyelinated club endings (which mediate field effect inhibition) penetrate within the peripheral part of the axon cap and terminate on this cell's soma and fine cap dendrites, while the small vesicle boutons which remain outside the axon cap (and can therefore only carry chemical inhibition) impinge on the Mauthner cell's soma and the proximal part of its large dendrites. An important finding was that endings of the two groups were stained by HRP, indicating that most commissural interneurons issue synaptic knobs of these two distinct classes. (2) The fine structure of labeled terminals was investigated: They exhibit the typical aspect of Gray type II synapses, with well‐differentiated presynaptic and postsynaptic differentiations and they contain pleiomorphic synaptic vesicles (mean diameter 40–46 nm; elongation coefficient 1.4–1.5; surface area 13–15.102 nm2). A comparison with adjacent unstained equivalent terminals has revealed that HRP allows a reliable analysis of synaptic organelles with, however, a slight tendency of vesicles to be more spherical in presence of the enzyme. (3) Spikes do not propagate actively in unmyelinated club endings but the remaining passively transmitted depolarization is nevertheless above threshold for inhibitory transmitter release, as indicated by unitary IPSPs evoked by fibers terminating exclusively within the axon cap. Thus, club endings have a dual action, and they mediate part of the M‐cell's chemical inhibition. (4) Synapses established by commissural cells on vestibular neurons contain a pleiomorphic population of vesicles; they can be compared to those which were identified in the vestibular nuclei of other species and which were also postulated to have inhibitory functions. (5) The synaptic investment of HRP‐stained commissural neuronal cell bodies included mixed synapses associating gap junctions and active zones established by presumably VIIIth nerve afferents. This observation can be related to the fast‐occurring inhibition produced in both M‐cells by vestibular nerve stimulation. (6) Axons of commissural fibers are myelinated and the largest of them have an internal diameter of no less than 10 μm; the ratio g (axon/total fiber) is close to the optimal value for maximum conduction velocity. Thus, again, these cells are likely to carry the nearly simultaneous ipsi‐lateral and contralateral inhibition of the M‐cell following VIIIth nerve excitation and their widespread distribution suggests a diffuse and synchronous ipsi‐lateral and contralateral inhibitory action on other nuclei as well. (7) In contradiction to electrophysiological results suggesting that some of these neurons were efferent to the labyrinth, a stained process was only observed once in the vestibular tract; this case confirms that commissural cells can project to the labyrinth but its rarity raises questions about the criteria commonly used for efferent identification.

Fine structure of the superficial layers of the viper optic tectum. A Golgi and electron-microscopic study.

The superficial layers of the viper optic tectum, which receive fibers from he retina, were studied using both light and electron microscopes. The optic fibers layer, or stratum opticum, is composed of 200 to 250 tight fascicles containing thin fibers, nearly all of which are myelinated. The main optic terminal layers, the stratum griseum et fibrosum superficiale, the greatest part of the cellular population is composed of small vertically oriented neurons and horizontal nerve cells, many of which are probably local circuit neurons. The neuropil of the stratum griseum et fibrosum superficiale is made up of small nerve elements, including three types of profiles containing synaptic vesicles; 1) boutons with pleiomorphic synaptic vesicles (P), representing over 47% of the total population of profiles containing synaptic vesicles and comprising three subgroups (P1, P2, and P3); 2) boutons with spheroidal synaptic vesicles (S), forming more than 29% of the total populations of profiles containing synaptic vesicles and comprising two categories, S1 and S2 (S2, the more numerous, represents the optic boutons, which make up 22% of the total populations of profiles containing synaptic vesicles); and 3) dendrites with pleiomorphic vesicles, accounting for approximately 23% of the total populations of profiles containing synaptic vesicles. A study of synaptic patterns revealed a large number of serial synapses and a lesser number of triplets or triadic synapses. The presynaptic components are boutons containing spheroidal (S1, S2) or pleiomorphic (P1, P2, P3) synaptic vesicles. The intermediate profile was always a dendrite with synaptic vesicles which frequently belonged to the small neurons of the stratum griseum et fibrosum superficiale. Comparison of the present results with other recent data shows that the synaptic circuitry in the optic tectum of Vipera aspis closely resembles the pattern observed in the optic tectum of other vertebrates, ranging form fish to mammals. However, quantitative differences exist, especially with regard to the proportion of dendrites containing synaptic vesicles. Their number seems to be higher in sauropsidians than in mammals, particularly in primates.

The bushy cells in the anteroventral cochlear nucleus of the cat. A study with the electron microscope

The bushy cells in the anterior division of the anteroventral cochlear nucleus of the cat were studied with the electron microscope. In the anterior part of the anterior division, profiles of bushy cells and their processes are easily identified, since few cells of other types are found in this region. In the posterior and posterodorsal parts of the anterior division, the bushy cells are intermingled with stellate and small cells but can be identified on the basis of light-microscopic descriptions and comparisons with the results from the anterior part. Bushy cells are large, spherical cells with a centrally located nucleus enveloped by sheets of rough endoplasmic reticulum. Thin proximal dendrites jut abruptly from the cell body and contain a relatively pale cytoplasm. The distal dendrites contain few organelles other than numerous, very large mitochondria. The cell soma and proximal dendrites, as well as the axon hillock, receive numerous synaptic terminals, but the distal dendritic processes are contacted by relatively few endings. At least four types of terminals form synaptic contacts with the bushy cells. Very large terminals, containing large, spherical synaptic vesicles and forming multiple asymmetrical contacts, correspond to the end-bulbs of Held from the cochlea. These terminals disappear after cochlear ablations, but the other three types remain. The most numerous of these is a large terminal that contains flattened synaptic vesicles and forms long, nearly symmetrical contacts with the soma and dendrites of bushy cells. The second type of non-cochlear terminal is smaller and contains small, pleiomorphic synaptic vesicles that are not flattened. The third type occurs mainly on bushy cell dendrites, contains small, spherical synaptic vesicles, and forms moderately asymmetrical contacts. The bushy cells probably correspond to the primarylike units described in electrophysiological studies of the anterior division. Primarylike units respond to activity in auditory nerve fibers in a one-to-one manner, a finding compatible with the observation that much of the surface of the soma and dendrites of the bushy cells is contacted by auditory nerve terminals (end-bulbs of Held). Neither the origins nor the functions of the several types of non-cochlear inputs to the bushy cells are known. Further analysis of these inputs and of the other neuronal types in the anterior division, when correlated with physiological and biochemical data from the same cell types, could clarify the functional significance of the observed patterns of synaptic organization.

The ventral lateral geniculate nucleus, pars lateralis of the rat. Synaptic organization and conditions for axonal sprouting.

The synaptic organization of the pars lateralis portion of the ventral lateral geniculate nucleus is similar to that of other thalamic nuclei. There are four types of synaptic knobs (RL, RS, F1, F2). RL knobs are large and irregularly shaped, contain round synaptic vesicles and make multiple asymmetrical junctions. They are found primarily in "synaptic islands" making contact with gemmules, spines, small dendrites, and other synaptic profiles containing pleiomorphic synaptic vesicles (F2). Smaller RS knobs contain round vesicles and make asymmetrical junctions with the same type of elements as RL knobs, with the exception of the F2 profiles, but are seldom found in synaptic islands. F1 knobs contain flattened synaptic vesicles and form symmetrical junctions with F2 knobs, gemmules, spines, and small-medium dendrites in synaptic islands, throughout the neuropil, and on the proximal dendrites and soma of the largest type of neuron. F2 knobs are irregularly shaped, contain pleiomorphic synaptic vesicles and make symmetrical junctions primarily with gemmules and spines in synaptic islands. They are postsynaptic to RL and F1 knobs. Occipital decortication indicates that cortical terminals are of the RS type. Bilateral enucleation indicates that retinal terminals are of both the FL and RS type. The large amount of geographic overlap of retinal and cortical terminals on gemmules, spines, and small dendrites found in the neuropil outside of synaptic islands logically would maximize axonal sprouting between these two sources.