Synaptic organization in the lateral geniculate nucleus of the monkey

Summary The synaptic organization in the lateral geniculate nucleus of the monkey has been studied by electron microscopy.The axon terminals in the lateral geniculate nucleus can be identified by the synaptic vesicles that they contain and by the specialized contacts that they make with adjacent neural processes. Two types of axon terminal have been recognized. The first type is relatively large (from 3–20 ) and contains relatively pale mitochondria, a great many vesicles and, in normal material, a small bundle of neurofilaments. These terminals have been called LP terminals. The second type is smaller (1–3 ), contains darker mitochondria, synaptic vesicles, and no neurofilaments. These have been called SD terminals.Both types of terminal make specialized axo-somatic and axo-dendritic synaptic contacts, but the axo-somatic contacts are relatively rare. In addition the LP terminals frequently make specialized contacts with the SD terminals, that is, axo-axonal contacts, and at these contacts the asymmetry of the membranes is such that the LP terminal must be regarded as pre-synaptic to the SD terminal.The majority of the synaptic contacts are identical to those that have been described previously (Gray, 1959 and 1963a) but, in addition, a new type of contact has been found. This is characterized by neurofilaments that lie close to the post-synaptic membrane, and by an irregular post-synaptic thickening. Such filamentous contacts have been found only where an LP terminal contacts a dendrite or a soma.The degeneration that follows removal of one eye demonstrates that the LP terminals are terminals of optic nerve fibres. The origin of the SD terminals is not known.The glial cells often form thin lamellae around the neural processes and tend to isolate synaptic complexes. These lamellae occasionally show a complex concentric organization similar to that of myelin.It is a pleasure to thank Prof. J. Z. Young for advice and encouragement and Dr. E. G. Gray for the considerable help he has given us. Dr. J. L. de C. Downer gave us much help with the care of the animals and with the operations. We also wish to thank Mr. K. Watkins for technical assistance and Mr. S. Waterman for the photography.     

Neuronal and synaptic organization of the lateral geniculate nucleus of the tree shrew,Tupaia glis

Summary The ultrastructural study of the lateral geniculate nucleus (LGN) of the tree shrew (Tupaia glis) revealed two types of neurons: (1) a large thalamocortical relay cell (TCR), which may bear cilia, and (2) a small Golgi type-II interneuron (IN) with an invaginated nucleus. The narrow rim of pale cytoplasm of the IN contains fewer lysosomes and fewer Nissl bodies than the cytoplasm of the TCR. The IN perikarya, which in some cases establish somatosomatic contacts, frequently contain flattened or pleomorphic synaptic vesicles. The ratio of TCR to IN is 31.Three types of axon terminals were observed in the LGN. Two of them contain round synaptic vesicles but differ in size. The large RL boutons undergo dark degeneration after enucleation; they are the terminals of retino-geniculate fibers. The smaller RS boutons show dark degeneration after ablation of the visual cortex; they are the terminals of the cortico-geniculate fibers. The third type of bouton (F₁ does not degenerate after either intervention. The boutons of this type are filled with flattened vesicles and are believed to be intrageniculate terminals. F₂-profiles were interpreted as presynaptic dendrites of the IN. The characteristic synaptic glomeruli found in the LGN contain in their center an optic terminal. These optic terminals establish synaptic contacts with dendrites or spine-like dendritic protrusions of TCRs as well as with presynaptic dendrites. Synaptic triads were also seen. The distribution of the individual types of synaptic contacts in layers 3 and 4 was determined. Layer 4 contains only one third of the retino-geniculate synapses and of the synaptic contacts of F₁-terminals.     

Synaptology of three peptidergic neuron types in the central nucleus of the rat amygdala

The synaptology of neurotensin (NT)-, somatostatin (SS)- and vasoactive intestinal polypeptide (VIP)-immunoreactive neurons was studied in the central nucleus of the rat amygdala (CNA). Three types of axon terminals formed synaptic contacts with peptide-immunoreactive neurons in the CNA: Type A terminals containing many round or oval vesicles; Type B terminals containing many pleomorphic vesicles; and Type C terminals containing fewer, pleomorphic vesicles. Peptide-immunoreactive terminals were type A. All three types of terminals formed symmetrical axosomatic and asymmetrical axodendritic contacts. However, type B and peptide-immunoreactive terminals frequently formed symmetrical axodendritic synaptic contacts. VIP-immunoreactive terminals also formed asymmetrical axodendritic contacts. SS- and NT-immunoreactive terminals commonly formed symmetrical contacts on SS- and NT-immunoreactive cell bodies, respectively. VIP-immunoreactive axon terminals were postsynaptic to nonreactive terminals. Type B terminals appeared more frequently on VIP neurons than on NT or SS neurons.     

Changes in signal transmission by neurons of the cat lateral geniculate nucleus by acute deafferentation and early degeneration.

1. The responses of single principal cells of the cat lateral geniculate nucleus (LGN) were recorded extracellulary from the optic radiation (OR) axons or intracellularly from the LGN. Electrical stimuli at different frequencies were applied to the optic tract (OT) to test the transneuronal and the synaptic signal transmission in the LGN. 2. The effect of acute deafferentation (by photocoagulation of the retinal receptive field) or of synaptic degeneration induced 2-4 days prior to the recording time on the LGN neuron signal transfer was studied. Immediately after deafferentation, the synaptic signal transfer by LGN neurons exhibits signs of hyperexcitability leading to multiple neuronal discharges. This acute deafferentation hyperexicitability is probably caused mainly by the disapperance of lateral inhibition mediated by LGN interneurons. The deafferentation hyperexcitability disappeared during electrical stimulation of the OT at frequencies greater than 10/sec. 3. With progressing degeneration of the synaptic terminals during the 2nd to 4th day after interruption of the optic nerve axoplasmic flow, the synaptic signal transfer by LGN neurons gradually deteriorates and ceases at the end of the fourth day. The signs of this deterioration (larger temporal scatter, increased exhaustability and reduced upper frequency limit of the transneuronal signal transmission and gradual reduction of the EPSP amplitude in D-neurons) were quantitatively investigated. 4. The neurophysiological data obtained at different levels of synaptic terminal degeneration are well correlated with morphological changes found within the degenerating synaptic terminals.     

The ventral lateral geniculate nucleus,pars lateralis of the rat

Summary 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 RL 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.We would like to thank Mr. Peter Rossetti for his excellent technical assistance on a major portion of this project, Ms. Judith Strauss for photographic assistance, and Ms. Nancy Wood for typing. Supported by grants NS 10579, NS 08724, 5 S01 RR 05402, and 2 T01 GM 00326     

Simultaneous Recording of Input and Output of Lateral Geniculate Neurones

TO understand the way in which the cat dorsal lateral geniculate nucleus (LGN) processes visual information it would be useful to know the number and type of retinal inputs to individual LGN neurones. Using electrical stimulation of the optic nerve Bishop et al.¹concluded that an impulse in a single optic nerve fibre is sufficient to excite a single LGN neurone. From the appearance of excitatory postsynaptic potentials (EPSPs) recorded essentially intracellularly, Creutzfeldt suggested that LGN neurones are driven by perhaps one² or a few³ retinal ganglion cells. Hubel and Wiesel⁴ proposed models of convergence of several retinal inputs on single LGN neurones based on analyses of receptive fields. Guillery⁵ produced anatomical evidence that some types of LGN neurones receive inputs from several different retinal fibres. Now we report direct observations which were made by recording simultaneously from single LGN neurones and from individual retinal ganglion cells which provided excitatory input to them. We shall not consider inhibitory influences, which are currently under study.     

Synapses in the cerebral ganglion of adult Ciona intestinalis

Summary Electron microscopy of the synaptic morphology of synapses in the cerebral ganglion of the adult ascidian (sea squirt) Ciona intestinalis reveals that the synapses are restricted to the central neuropil of the ganglion. Many of the synapses show a polarity of structure such that pre and post synaptic parts can be identified. The vesicles in the presynaptic bag are of two main diameters 80 and 30 nm respectively. The large vesicles have electron dense contents that vary both in their capacity and dimensions.The pre and postsynaptic membranes are more electron dense than the surrounding membranes, but they are only slightly thicker. Both the pre and post synaptic membranes have electron dense dots some 10 nm in diameter associated with their cytoplasmic surfaces. Sometimes the presynaptic membrane has larger peg-like projections between the vesicles. Associated with the post synaptic membrane are tubules some 10 nm in diameter. These tubules may be the dots cut obliquely.The synaptic cleft material is more electron dense than the surrounding intercellular material, and in it there is a dense line made up of granules about 3–5 nm in diameter. This dense line is usually mid way between the pre and post synaptic membranes, but may be nearer the postsynaptic membrane.No tight junctions between adjacent nerve process profiles have been observed.I wish to thank Professors J. Z. Young, F. R. S. and E. G. Gray for much advice and encouragement, also Dr. R. Bellairs for the use of electron microscope facilities and Mr. R. Moss and Mrs. J. Hamilton for skillful technical assistance.     

Electron microscopic observations of synaptic endings in cultures of mammalian central nervous tissue

Summary Synapses were found in rat cerebellar and brainstem cultures with the electron microscope. Three distinct types of synaptic terminals were described. The similarity between synapses found in vitro and in vivo was emphasized.Supported by USPHS Grants 5 Tl 459-04 and NB 03114-03S1 from the National Institutes of Health, Bethesda, Maryland.The authors wish to express their sincere appreciation to Mrs. Eleanor Morris for her assistance in preparing the cultures and Mr. Earl Pitsinger for his photographic assistance.     

Observations on the cellular localization of dopamine in the caudate nucleus of the rat

Summary The cellular localization of dopamine in the caudate nucleus of the rat hat been studied with the highly sensitive and specific fluorescence method of Falck and Hillarp, and by electron microscopy. The histochemical studies provided strong support for the view that the dopamine is concentrated within very fine nerve fibres which have abundant varicosities with an intense fluorescence. The electron microscopical studies revealed the presence of a tightly packed plexus built up i.a. of abundant synaptic nerve terminals, many of which had a diameter below 0.4 . The terminals made synaptic contact mainly with processes that seemed to belong to an extensive dendrite net.The investigation was supported by research grants from the United States Public Health Service (02854-04), The Swedish Medical Research Council and the Knut and Alice Wallenberg Foundation.     

Osmium tetroxide-zinc iodide reactive sites in the photoreceptor cells of the retina of the rat

Summary The osmium tetroxide-zinc iodide fixative of Champy-Maillet has been used to study the rat's retina at the electron microscope level. Electron opaque deposits were observed all along the photoreceptor cells and concentrated in the outer segments of rods and cones and in the nerve endings. In the outer segments that deposits are located in the inter and intra disk spaces as well as between the disk and outer membranes. In the outer plexiform layer reactive sites include synaptic vesicles and mitochondria; other minor reactive sites are described in the inner segment and inner plexiform layer.Electron opaque deposits were not seen if potassium iodide substitutes zinc iodide in the fixative. However, if osmium tetroxide-potassium iodide fixed retinae are immersed in osmium tetroxide-zinc iodide the characteristic electron-dense material is evidenced at those same sites. The effect of other several fixatives were studied with a similar double fixation procedure. Our finding points to the histochemical demonstration of an unidentified component (s) of the retina which shows a striking specificity of localization and which is made evident when zinc iodide is used in the Champy-Maillet mixture.This work has been supported by grants of the Consejo Nacional de Investigaciones Cientificas y Técnicas, Argentina and U.S. Air Force AF-AFOSR 67-0963 A.We are greatly indebted to Miss Haydée Agoff and to Mr. Alberto Saenz for their skillful technical assistance.     

Photoperiodic and osmotic influences on the ultrastructure of the hypothalamic neurosecretory system of the White-crowned sparrow,Zonotrichia leucophrys gambelii

Summary An attempt was made to correlate functional changes in the neurohypophysis of the White-crowned Sparrow, Zonotrichia leucophrys gambelii, with morphologic features on the light- and electron-microscope levels. The aldehyde-fuchsin-staining anterior median eminence possesses essentially the same ultrastructural features as the non-staining posterior median eminence. The axon terminals are characterized by the presence of a large number of small vesicles (approximately 400 Å in diameter) and occasional electron-dense granules. The more-or-less depleted anterior median eminence occasionally evident in the photosensitive bird showing testicular development is indistinguishable ultrastructurally from the more intensely staining median eminence generally characteristic of the photorefractory bird. In the median eminence, stainability and functional state do not seem to be correlated with changes in the type, size or number of vesicles. A slight increase in the number of granules was noted in the photorefractory bird but this was considered insufficient basis to account for differences in stainability.The pars nervosa, on the other hand, responded to osmotic stimuli (saline drinking water) by loss of stainability and decrease in numbers of elementary neurosecretory granules. Small vesicles are also present in the pars nervosa axon terminals, but are intermingled with neurosecretory granules in normal birds. Acute-osmotic birds, however, had axon terminals almost entirely occupied by small vesicles.It is to be emphasized that the pars nervosa and the median eminence are two structurally very different regions of the neurohypophysis. The basis for aldehyde-fuchsin staining in the median eminence appears to differ from that in the pars nervosa. The implications of these findings are considered in regard to hypothalamic control over gonadotropic activity in the White-crowned Sparrow.Dedicated to Professor Dr. W. Bargmann in honor of his 60th birthday.This investigation was supported by grant GB-2484 from the National Science Foundation to Professor Bern, grant GB-2819 from the National Science Foundation to Professor Mewaldt, and grant NB-01353 from the National Institutes of Health to Professor Farner. The authors wish to express their appreciation of the technical assistance of Mrs. Irene Brown, Mr. John Butchart, Sally S. Kibby, Mrs. Carol Nicoll, and Mr. John Striffler. Mrs. Emily Reid kindly prepared the histograms.     

On the neurons and synapses of the lateral geniculate nucleus of the monkey,and the effects of eye enucleation

Summary Two neuron types are distinguished by electron microscopy in the lateral geniculate nucleus (LGN) of the monkey-a large cell (P cell) interpreted as a geniculostriate relay cell, and a small cell (I cell) interpreted as an inhibitory interneuron. The I cell, distinguished by its small size, infolded nucleus, small mitochondria, cilium and small granular bodies, forms about 10% of the total neuron population. It could not be determined whether this cell has an axon, but its dendrites, which contain aggregates of flattened vesicles, are thought to form a proportion of the F processes, profiles which are post-synaptic to the retinal (RLP) axons and presynaptic to the dendrites of the P cells. The small dark (RSD) axon terminals of unknown origin contact the dendrites of both cell types.After eye enucleation the P cells of the affected laminae of the LGN shrink and partially withdraw their dendrites from the neuropil. By 29 months' survival, they have only a narrow cytoplasmic rim around the nucleus. A necrotic process also occurs, affecting fine dendrites by 22 days and large profiles by 45 days, but it is not clear whether whole cells are destroyed by this process. At 45 days the I cells are commonly seen to form somatodendritic synapses. The appearance of these synapses is interpreted as the result of a withdrawal to the soma of the presynaptic dendrites.It is concluded that the I cells are probably inhibitory interneurons subject to excitation and presynaptic inhibition by the RLP and RSD axons, and a diagram is presented to demonstrate the possible significance of these connections for the transmission of information through the LGN.The author wishes to thank Dr. J. Campos-Ortega for much practical advice.     

The structure of a photoreceptor organelle in the eye of Pterotrachea mutica

Summary The photoreceptor cell of Pterotrachea consists of an elongated cell some 100 m long with recogniseable inner and outer segments. The photoreceptor membranes point towards the light. There are about 300 discs per photoreceptor, a small number of discs arising from a single ciliary base. There are bout 75–100 such bases on each receptor cell. The receptor cells themselves (the inner segments) have four recognisable regions. The vacuolated region, the region of mitochondria, the nuclear region, and the axonal region.The photoreceptor cells are organised in five roughly parallel rows, and separated from one another by pale supporting cells.My thanks are due to Professor J. Z. Young, F. R. S. for his enthusiastic support and help during this work. Dr. R. Bellairs kindly provided electron microscope facilities. Mr. R. Moss, Mrs. J. Hamilton and Mr. A. Aldridge provided excellent technical and photographic assistance.     

Synaptogenic effect of estrogen on the hypothalamic arcuate nucleus of the adult female rat

Summary In order to examine the effect of estrogen on the synaptic structures in the hypothalamic arcuate nucleus (ARCN), semi-quantitative studies were performed by counting synapses in an 18,000 m² area in the middle part of the ARCN in each brain. In ovariectomized female rats injected with 2 g of estradiol benzoate (EB) for three weeks, the mean numbers of axodendritic and axosomatic synapses were not significantly different from those in the intact and ovariectomized controls. When the medial basal hypothalamus (MBH) including the ARCN was isolated by use of a Halász knife (MBH island), the mean number of axodendritic synapses was decreased to about half of the controls. However, EB treatment for three weeks from the day of surgery effectively restored the axodendritic synaptic population of the deafferented ARCN. This may suggest that estrogen has a facilitatory effect on axodendritic synapse formation in the deafferented ARCN, presumably by stimulating axonal sprouting and synaptic regeneration of intact axons in the MBH island.Supported by grants from the Ministry of Education of Japan     

Histochimie des mono-amine-oxydases du tronc cerebral du rat: noyaux latero-dorsal du tegmentum et interpedonculaire

Résumé L'étude histochimique des Mono-Amine-Oxydases (MAO) des noyaux gris du tronc cérébral: interpédonculaire et latéro-dorsal du tegmentum a été réalisée chez 30 rats albinos mâles et femelles.La méthode de Glenner avec les sels de Tétrazolium, modifiée par Shimizu, a été employée et nous a permis de confirmer les travaux de ces auteurs et ceux de Smith.Le noyau latéro-dorsal du tegmentum paraît avoir la plus forte teneur en MAO du cerveau; sa coloration est dense, homogène, diffusant légèrement autour du site observé lors de l'étude histologique par la méthode d'Einarson à la Gallocyanine.Au contraire, celle du noyau interpédonculaire est moins intense, non homogène, plus marquée latéralement.L'action in vivo des inhibiteurs spécifiques des MAO (Phénelzine, Nialamide) confirme la disparité de ces deux noyaux: la dose minimale efficace pour inhiber totalement les MAO du n. interpédonculaire est insuffisante pour le n. latéro-dorsal du tegmentum.La Phénelzine a une action beaucoup plus précoce et plus complète que le Nialamide; leurs durées d'action sont voisines.

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Summary The histochemical study of the Mono-amine-oxidases (MAO) in the nuclei interpeduncularis and latero-dorsalis tegmenti of the brain stem was made over 30 male and female albinos rats.The Glenner's method with Tetrazolium salts, modified by Shimizu, was used and enabled us to confirm the works of these authors and Smith as well.The coloration of the nucleus latero-dorsalis tegmenti is of a strong and homogeneous density and spreading slightly all around the spot observed during the histological study with Gallocyanin after Einarson's method.On the contrary, the coloration of the nucleus interpeduncularis is less strong, without any homogeneity, and of thicker density on the edges.The activity in vivo of the specific inhibitors of MAO (Phenelzin, Nialamid) confirms the disparity of their 2 nuclei. Phenelzin operates earlier and more completely than Nialamid. The time of their action is similar.

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Remerciements. Nous remercions Meile Gontcharoff (lab. Biologie Générale) et Mr Jacquot (lab. Physiologie Animale) de la faculté des Sciences de Reims, qui nous ont acceuilli généreusement dans leur laboratoire et nous ont permis de réaliser cette étude. Nous exprimons notre gratitude envers Mr Jouvet (lab. Médecine expérimentale, faculté de Médecine, Lyon) qui nous a adressé ses critiques.Nos remerciements vont aussi à Mr Lechenault, Maître-Assistant de Biologie Générale (Fac. des Sciences, Reims) qui nous a prodigué ses conseils compétents et sans qui cette étude n'aurait pu être menée à bien.     

The ultrastructural changes in the neurohypophysis after destruction of the paraventricular nuclei in normal and castrated rats

Summary In normal and castrated rats an electrolytic lesion of 1–1.5 mm in diameter was placed in the hypothalamus using bilateral electrodes. Such a lesion destroyed both paraventricular nuclei and surrounding tissue. The electron microscope study of the neurohypophysis revealed that about 1/5 of the neurosecretory axons and terminals degenerated. In the castrated rats, the clear axons described in a previous paper (Zambrano andDe Robebtis, 1968), were those that underwent degeneration. This finding supports our previous assumption that such axons belong to the paraventricular system. The sequence of the ultrastructural changes occurring in the degenerating axons and terminals is described. Special features were the early lysis of the neurotubules, the breakage of the membrane of the elementary granules and of the axolemma. Disrupted axonic material was observed in the interstitial tissue. The degeneration of the clear axons from the paraventricular system is discussed in relation to their possible oxytocinergic nature.An early cellular reaction was found in the perivascular microglial type of cell. These increased in number and underwent ultrastructural changes indicative of active phagocytosis and pinocytosis. Both the protoplasmatic and the fibrous pituicytes did not participate in the removal of the degenerating axons. This was apparently done by extracellular digestion and engulfment by microglial cells.Supported by grants from the Consejo Nacional de Investigaciones Científicas y Técnicas and by the Air Force Office of Scientific Research (AF-AFOSR 963-67).The Authors are grateful to Dr.S. Taleisnik, Instituto de Investigaciones Médicas Mer cedes y Martín Ferreira, Córdoba, Argentina for encouraging this investigation and for the technical help so generously offered.     

The synaptic organization in the medial geniculate body of afferent fibres ascending from the inferior colliculus

Summary The ventral nucleus of the medial geniculate body has been examined electron microscopically 2–5 days after destruction of the inferior colliculus. In both the ipsi- and contralateral ventral nuclei, degenerating collicular afferents are of medium diameter (1–5 ) and their degenerating terminals are distributed mainly to synaptic aggregations (glomeruli) in which they end axo-axonically and axo-dendritically. Their distribution and mode of termination indicates that these terminals belong to a class which in normal material is large, contains round synaptic vesicles and ends by means of asymmetrical synaptic complexes upon dendrites and upon the second (pale) type of glomerular terminal. It also ends by means of adhesion plaques on the same dendrites.As the terminals of corticothalamic afferents to the nucleus are already known, only the origin of two types remains to be determined: the pale terminals, which arise from structures resembling dendrites and which end only axo-dendritically, and a small, less common terminal which ends axo-axonically, axo-dendritically and axo-somatically. Both types contain flattened synaptic vesicles and end by means of symmetrical synaptic complexes.Correlative Nauta and Golgi studies suggest that the collicular afferents have a very specific spatial distribution within the cellular laminae composing the ventral nucleus.The terminal degeneration commences as a neurofilamentous hyperplasia and quickly passes to one of increased electron density. There is evidence for early removal of degenerating terminals from the postsynaptic membrane.This work was supported by a grant from the Bank of New Zealand Medical Research Fund.We are indebted to Professor J. A. R. Miles for use of the electron microscope.     

An electron microscopical study of the ventral nerve cord of the leech

Summary Three types of fibrillar structure can be seen with the electron microscope in nerve cells of the vental nerve cord of the leech: the neurofibrillar bundles, the tubules and the tonofibrils. In neuroglial cells only the tonofibrils are present. The three types are structurally distinct, and, contrary to past suggestions, there is no evidence that neurofibrillar bundles may consist of tightly packed or badly fixed tubules.In vertebrates the electron microscope reveals bundles of discrete neurofilaments that form the basis for the argyrophilic neurofibrillae seen by light microscopy. Each neurofilamentous unit appears as a dot in cross section. In contrast, in the leech, the electron microscope shows compact fibrillar bundles that clearly correspond to the neurofibrils described by light microscopists. These bundles are made up of closely packed units rather than discrete filaments and where the units occur singly they are seen to have an angular or stellate outline in cross section. To make this distinction clear these have been termed neurofibrillar bundles rather than neurofilaments.Attachment plaques occur in both neurons and neuroglia. These plaques have tonofibrils attached, and the glial tonofibrils are far more numerous than the neuronal tonofibrils. The glial fibrils are identical with the tonofibrils in the glial cells.The attachment plaques are invariably related to an extracellular space that contains material identical with the basement membrane. This material is continuous, by a complex system of channels and diverticulae, with the outer basement membrane in the neuron packets, but forms isolated patches in the other parts of the nervous system.We are grateful to Prof. J. Z. Young, F. R. S., for his encouragement to Mrs. Astafiev for the drawings, to Miss B. Shirra and Mr. K. Watkins for technical assistance and to Mr. S. Waterman for photography.     

Synaptic relationships in the plexiform layers of carp retina

Summary The synaptic contacts made by carp retinal neurons were studied with electron microscopic techniques. Three kinds of contacts are described: (1) a conventional synapse in which an accumulation of agranular vesicles is found on the presynaptic side along with membrane densification of both pre- and postsynaptic elements; (2) a ribbon synapse in which a presynaptic ribbon surrounded by a halo of agranular vesicles faces two postsynaptic elements; and (3) close apposition of plasma membranes without any vesicle accumulation or membrane densification.In the external plexiform layer, conventional synapses between horizontal cells are described. Horizontal cells possess dense-core vesicles about 1,000 Å in diameter. Membranes of adjacent horizontal cells of the same type (external, intermediate or internal) are found closely apposed over broad regions.In the inner plexiform layer ribbon synapses occur only in bipolar cell terminals. The postsynaptic elements opposite the ribbon may be two amacrine processes or one amacrine process and one ganglion cell dendrite. Amacrine processes make conventional synaptic contacts onto bipolar terminals, other amacrine processes, amacrine cell bodies, ganglion cell dendrites and bodies. Amacrine cells possess dense-core vesicles. Ganglion cells are never presynaptic elements. Serial synapses between amacrine processes and reciprocal synapses between amacrine processes and bipolar terminals are described. The inner plexiform layer contains a large number of myelinated fibers which terminate near the layer of amacrine cells.This work was supported by an N.I.H. grant NB 05404-05 and a Fight for Sight grant G-396 to P.W. and N.I.H. grant NB 05336 to J.E.D. The authors wish to thank Mrs. P. Sheppard and Miss B. Hecker for able technical assistance. P.W. is grateful to Dr. G. K. Smelser, Department of Ophthalmology, Columbia University, for the use of his electron microscope facilities.     

Electron microscopic and histochemical observations on different types of nerve endings in the extraocular muscles of the rat

Summary Nerve endings in the extraocular muscles of the rat were submitted to histochemical tests for formalin-induced fluorescence and carboxylic esterases. Acetylthiocholine, butyrylthiocholine and -naphthyl acetate were used as substrates and iso-OMPA, 284C51, eserine and E-600 as inhibitors. The ultrastructure of the endings was studied with the electron microscope.Both single and multiple nerve terminals were observed in all six extraocular muscles. The single terminals of myelinated axons were comparable in their light and electron microscopic structure with the typical motor end plates of other striated muscles, and like these they exhibit acetylcholinesterase (AChE), non-specific cholinesterase (ns. ChE) and non-specific esterase (ns. E) activity. These endings were apposed to twitch-type muscle fibres.The multiple terminals were classified with the light microscope into two types. The larger type was 1/3 of the size of the motor end plate; 2–5 endings innervated the same muscle fibre; subneural infoldings were weakly developed and possessed only slight AChE and ns. ChE and probably no ns. E activity. No subneural lamellae were visible under the light microscope in the smaller type, which also possessed AChE and ns. ChE and was composed of 10–20 small dots dispersed along a single muscle fibre. The Schwann cells along nerve fibres leading to these two types of multiple endings exhibited ns. ChE but not AChE and ns. E activity.The ultrastructure of the two types of multiple endings was principally similar. The main difference, compared with the motor end plate, was that these endings were derived from unmyelinated axons which either make synaptic contacts along their course with the muscle fibre at variable distances (smaller-type) or these terminals were grouped closely together (larger-type).A few dense-core vesicles were observed in these unmyelinated nerves and in their terminals which were considerably smaller than those in the motor end plate. They were not always separated from each other by sarcoplasm and teloglia (larger-type) and contained also empty vesicles. The secondary synaptic clefts were often sparse and irregular or even absent, but the typical myoneural postsynaptic electron density was always observed. These multiple endings, in contrast to the motor end plate, were apposed only to muscle fibres with slow contraction.No catecholamine containing nerve endings were observed in the extraocular muscles. These observations indicate that the rat extraocular muscles have a double cholinergic innervation.The author wishes to express his gratitude to Professor Antti Telkkä, M. D., Head of the Electron Microscope Laboratory, University of Helsinki, for permission to avail himself of the electron microscope facilities.