Fine structure of smooth muscle and neuromuscular junctions in the foot of Helix aspersa

Summary The smooth muscle cells in the foot of Helix aspersa are arranged in bundles which interweave to form a complex mesh. In the peripheral cytoplasm of the muscle cells there is a system of interconnected obliquely and longitudinally orientated tubules. The full extent of this system has not been determined; its possible function in relation to Ca⁺⁺ storage and excitation-contraction coupling is discussed. Longitudinal tubules are present among the myofilaments and in association with mitochondria. Distributed throughout the myofilaments are elliptically shaped dense bodies, the fine structure of which resembles an accumulation of thin filaments. Located on the plasma membrane of the muscle cells are dense areas; the fine structure and relationships of these cellular elements resemble desmosomes. They may serve as attachment points for thin, cytoplasmic filaments (not necessarily myofilaments). The muscle cells are innervated by axons which diverge from a coarse, neural plexus (the sole plexus). The axons initially come into close contact with the muscle cells and then pass over their surfaces for up to 35 before being gradually enveloped by flange-like protrusions of the muscle cells. These axons contain either, (i) agranular vesicles (600 Å in diameter), (ii) agranular and very dense granular vesicles (1000 Å in diameter) or (iii) agranular and less dense, granular vesicles (1000 Å in diameter). The possible role of these inclusions as sites of excitatory and inhibitory transmitters is discussed.I wish to thank Professor G. Burnstock for making laboratory facilities available. This work has been supported by the Australian Research Grants Committee.     

Ultrastructure of the neurohaemal region of the toad median eminence

Summary According to the internal structure and size of the granules, six types of nerve endings can be distinguished in the toad median eminence: 1. Endings containing mostly dense granules of 600 Å in diameter; 2. Endings containing dense granules of about 800 Å in diameter; 3. Endings which contain dense granules 1,000–2,000 Å in diameter, with the peak at 1,200–1,400 Å; 4. Endings containing granules with a characteristic structure, which differentiate them from the other three types; 5. Scarce endings containing granules 2,000 to 3,800 Å in diameter; and 6. Endings containing only vesicles 400–500 Å in diameter. Types 3 and 4 endings are mainly found in the outer pericapillary zone, and are probably responsible for the strong Gomori-positive reaction observed in this zone. The other four types of endings occur mainly in the inner pericapillary zone, and appear to be Gomori-negative.The probable origins of the different types of endings, and their possible relations with the different releasing factors is discussed.The subendothelial basement membrane has numerous long processes which form a complicated network in contact with all the nerve endings, some nerve fibres and glial cells.Two types of glial cells are described. Pinocytotic vesicles are frequently seen at the points where these cells contact the basement membrane. All the ultrastructural features suggest that these cells are carrying out transport functions.Fellow of the Consejo Nacional de Investigaciones Científicas y Técnicas de la República Argentina.The author is very grateful to Professor H. Heller for his continued encouragement and criticism and to Mr. J. Lane and Mr. P. Heap for their valuable help.     

Electron microscopic study on the innervation of the pancreas of the domestic fowl

Summary The innervation of the pancreas of the domestic fowl was studied electron microscopically. The extrapancreatic nerve is composed mostly of unmyelinated nerve fibers with a smaller component of myelinated nerve fibers. The latter are not found in the parenchyma. The pancreas contains ganglion cells in the interlobular connective tissue. The unmyelinated nerve fibers branch off along blood vessels. Their synaptic terminals contact with the exocrine and endocrine tissues. The synaptic terminals can be divided into four types based on a combination of three kinds of synaptic vesicles. Type I synaptic terminals contain only small clear vesicles about 600 Å in diameter. Type II terminals are characterized by small clear and large dense core vesicles 1,000 Å in diameter. Type III terminals contain small clear vesicles and small dense core vesicles 500 Å in diameter. Type IV terminals are characterized by small and large dense core vesicles. The exocrine tissue receives a richer nervous supply than the endocrine tissue. Type II and IV terminals are distributed in the acinus, and they contact A and D cells of the islets. B cells and pancreatic ducts are supplied mainly by Type II terminals, the blood vessels by Type IV terminals.This work was supported by a scientific research grant (No. 144017) and (No. 136031) from the Ministry of Education of Japan to Prof. M. Yasuda     

Electron microscopical demonstration of acid phosphatase in blood platelets

Summary Ultrastructures of human and rabbit thrombocytes reveal specific subcellular organelles within these elements. Serotonin granules are demonstrated containing extremely electron opaque material in vesicles with an average diameter of 1,700 Å and a considerable number of large dense bodies (average size 4,000 Å in diameter) is seen. The latter are less electron dense as compared to the serotonin granules. The appearance of serotonin granules in the human thrombocyte is rare, while rabbit platelets show a higher number of these granular vesicles.Acid phosphatase activity in the large dense bodies of human and rabbit platelets has been demonstrated by means of electron microscopy. Present results together with currently available biochemical information are briefly discussed in relation to the lysosomal activity within the thrombocytes.     

The neurohypophysis of the crested newt

Summary In the median eminence of the newt a medial region and two lateral regions are described.In cross section, the medial region appears to be made up of 1) an outer or glandular zone (Zone I) containing aldehyde-thionine-positive and negative nerve fibres and blood capillaries. Nerve fibres appear aligned in palisade array along the capillaries. 2) An inner zone (Zone II) made up of a) a layer of aldehyde-thionine-positive nerve fibres (fibrous layer) belonging to the preoptic hypophyseal tract and b) a layer of ependymal cells lining the infundibular lumen and reaching the blood vessels with their long processes.The lateral regions display a less pronounced stratification and aldehyde-thionine positive nerve fibres are nearly absent.A slender lamina (ependymal border) containing mainly aldehyde-thionine-positive nerve fibres and ependymal cells connects the median eminence to the pars nervosa.At the ultrastructural level, in the outer zone of the medial region at least 4 types of nerve fibres and nerve endings are identified:Type I nerve fibres containing granular vesicles of 700–1000 Å and clear vesicles (250–400 Å).Type II nerve fibres containing granular vesicles and polymorphous granules of 900–1300 Å and clear vesicles (250–400 Å).Type III nerve fibres containing dense granules of 1200–2000 Å and clear vesicles of 250–400 Å.Type IV nerve fibres containing only clear vesicles of 250–400 Å. In the inner zone too, all these nerve fiber types are found among ependymal cells, while the fibrous layer consists of nerve fibres containing granules of 1200–2000 Å in diameter.In the lateral regions Type I, Type II and Type IV nerve fibres and their respective perivascular terminals are found; axons containing dense granules (1200–2000 Å) are scanty. In these regions typical synapses between Type I nerve fibres and processes rich in microtubules are visible.The classification and functional significance of nerve fibres in the median eminence are still unsolved, but it may be assumed that nerve fibres of the medial region belong to both the preoptic hypophyseal and tubero hypophyseal tract, while the lateral regions are characterized by nerve fibres of the tubero hypophyseal tract. Peculiar specializations of the ependymal cells in the median eminence of the newt are also discussed.Work supported by a grant from the Consiglio Nazionale delle Ricerche.The authors are indebted to Mr. G. Gendusa and P. Balbi for technical assistance.     

The small pyramidal neuron of the rat cerebral cortex

Summary The pyramidal neurons in layers II and III of the rat parietal cortex have dendritic spines which form synapses with axon terminals. These synapses have synaptic clefts containing granular material that is concentrated towards the middle of the cleft to form a plaque. Only a small amount of dense material occurs on the cytoplasmic face of the presynaptic membrane, while there is a prominent dense layer, some 300 Å deep, in the dendritic spine. When the synapses formed by the smallest dendritic spines are examined in a frontal or en face plane of section this postsynaptic density has the form of a disc. In the synapses on larger spines, the disc is perforated to form a ring, and in the largest spines a number of perforations may occur. Because of these perforations, in larger synapses sections passing at right angles to the plane of the synaptic junction may show two or more separate postsynaptic densities. The possible significance of these findings is discussed.This work was supported by United States Public Health Service Research Grant No. NB-07016 from the National Institutes of Neurological Diseases and Blindness. The authors wish to express their sincere thanks to Lawrence McCarthy and Charmian Proskauer for their valuable assistance.     

The fine structure of ependymal cells part II: An electron microscopic study of the soft-shelled turtle paraventricular organ,with special reference to the fine structure of ependymal cells and so-called albuminous substance

Summary The present study deals with the electron microscopic observations on the softshelled turtle paraventricular organ, with special reference to the relationship of the ependymal cells and the so-called albuminous substances. It is shown that the so-called albuminous substances consist of the tips of neuronal processes extending into the ventricular lumen. They probably arise from the nerve cells lying within the hypendymal or the underlying tissue. The ependymal cells of the PVO themselves are basically similar in structure to those of any other animal.The processes observed contain two types of vesicles, namely: the clear vesicle, 500 to 1600 Å in diameter, and the cored vesicle, measuring 600 to 1500 Å in diameter, which has a distinct membrane enclosing an extremely dense core of variable sizes. The functional significance of these vesicles is discussed in relation to that of inclusions in the neurosecretory and the autonomic nerve fibers in the hypothalamus.The findings indicate that in the terminal endings of the processes a production or formation of vesicles might occur and that these vesicles might be discharged into the cerebrospinal fluid by microapocrine secretion.The author's grateful thanks are due to Prof. E. Yamada for his continuously kind guidances and due to Prof. T. Sakurai for his constant encouragement.     

Morphology of microtubules in rabbit platelets

Summary The morphology of the microtubular wall in rabbit platelets fixed in glutaraldehyde and osmic acid solution and stained with uranyl acetate, or lead hydroxide, or doubly stained, is variable. In cross section, the wall may appear as a uniformly dense annulus, an annulus containing nodular densities about 35 Å in diameter or as a series of contiguous subunits with a circular cross sectional profile about 70 Å wide. The authors relate the varying morphology to the intensity of staining and equate the nodular and circular subunits. Rotational analysis suggests that there are 12 ± 2 subunits in the microtubular wall.We thank Professor A. C. Ritchie for his criticism of the paper and Mrs. M. Lorber and Mrs. M. Mezari for technical assistance. — This work was supported by grants from the Medical Research Council of Canada and the Ontario Heart Foundation.     

Auerbach's plexus of mammals and man: Electron microscopic identification of three different types of neuronal processes in myenteric ganglia of the large intestine from rhesus monkeys,guinea-pigs and man

Summary Ganglia from Auerbach's plexus of the large intestine (caecum, appendix vermiformis, colon transversum and rectum) in man, rhesus monkey and guinea-pig are composed of nerve cells and their processes, typical Schwann cells and a vast neuropil. The neuropil consists of dendrites and axons of intrinsic nerve cell perikarya and axons of extrinsic neurons. Axonal profiles in large nerve fibre bundles are of uniform size and appearance, embedded in infoldings of Schwann cell cytoplasm and contain occasional large granular vesicles, mitochondria and neurotubules. Preterminal axons widen into vesicle filled varicosities, some of which establish synaptic contact with intrinsic nerve cell bodies.At least three different types of neuronal processes can be distinguished in the myenteric neuropil according to the size, appearance and commutual proportion of vesicles present in axonal varicosities, and their ability to accumulate exogenous 5- and 6-hydroxydopamine and 5-hydroxydopa: 1. Axonal enlargements containing a major population of small electron lucent synaptic vesicles (350–600 Å in diameter) together with a small number of membrane-bound, opaque granules (800–1,100 Å). These profiles have been identified as cholinergic axons. The boutons establish synaptic contacts with dendritic processes of intrinsic nerve cell bodies; membrane specializations are found at the preand postsynaptic sites. 2. Axonal beads of sometimes very large diameter, containing an approximately equal amount of large granular vesicles (850–1,600 Å) and small, electron lucent or faintly opaque vesicles (400–600 Å). The granular core of the large vesicles is of medium electron density and may either fill the entire vesicle or is separated from the limiting membrane by a more or less clear interspace. The fibres probably belong to intrinsic neurons, and because of the similarity of the large, membrane-bound vesicles with neurosecretory elementary granules, they have been designated p-type fibres (polypeptide fibres). The granular core of the vesicles in these fibres becomes more electron dense after treatment with 5-OH-dopa. The accumulation of an amine precursor analogue in combination with a possible storage of a polypeptide substance (or an ATP-like substance) resembles the situation in several diffusely distributed endocrine cell systems. 3. Varicosities of axons equipped with small (400–600 Å) empty or sometimes granular vesicles, medium sized (500–900 Å) vesicles with highly electron dense cores and occasional large (900–1,300 Å) granular vesicles. Pretreatment with 5-OH-dopamine increases the electron density in almost all medium-sized granular vesicles and some of the large granular vesicles; an osmiophilic core develops in some small vesicles. 6-hydroxydopamine results in degenerative changes in the varicosities of this type of neurons. Concomitantly, both catecholamine analogues markedly reduce neuronal noradrenaline in the large intestine, as demonstrated by fluorescence histochemistry and in fluorimetric determinations. The ultrastructural features of these varicosities and their reaction to 5- and 6-OH-dopamine indicate that they belong to adrenergic, sympathetic nerves. No membrane specializations could be detected at sites of close contact of the adrenergic boutons with dendrites and cell bodies of intrinsic nerve cells.Supported by grants from the Deutsche Forschungsgemeinschaft.Supported by a grant from Albert Pahlsson's Foundation, Sweden. The work was carried out within a research organization sponsored by the Swedish Medical Research Council (projects No. B70-14X-1007-05B, B70-14X-712-05, and B70-14X-56-06).     

Fine structure of smooth muscle and neuromuscular junctions in the optic tentacles of Helix aspersa and Limax flavus

Summary The smooth muscle cells studied contain a central core of thick and thin myofilaments surrounded by a peripheral layer of myofilament-free cytoplasm. Numerous vesicles, tubules, microfilaments, mitochondria and fine granules are present in the peripheral cytoplasm. Glycogen particles are distributed in large or small groups in both the peripheral cytoplasm and among the myofilaments. In contracted muscle cells the peripheral cytoplasm bulges out at regular intervals into the intercellular connective tissue. Numerous close contacts between single, usually naked, axons and these cytoplasmic protrusions occur. The axons at these contacts contain numerous small (500 Å in diameter) and large vesicles (800–1000 Å in diameter). Sometimes a number of axons simultaneously form close contacts with a muscle cell. These close contacts are considered to be the sites at which transmitter is released and acts on the muscle cell membrane.I wish to thank Professor G. Burnstock for making laboratory facilities available. This work has been supported by the Australian Research Grants Committee.     

Electron microscopic identification of the interphase chromosomes of Amoeba proteus and Amoeba discoides using autoradiography; with some notes on helices and other nuclear components

Data obtained from light and electron microscope autoradiographs of cells of Amoeba proteus and Amoeba discoides previously incubated in medium supplemented with H³ thymidine, indicate that fibrous material, the basic unit of which is about 150 Å in diameter, represents the interphase chromosomes of these amoebae. The helices of interphase nuclei do not appear to incorporate H³ thymidine, which is in opposition to the hypothesis of Taylor (1963) that they are G₂ chromosomes, and makes it unlikely that they represent any form of the DNA-containing component of the amoeba's interphase nucleus. Stereo-electron microscopy reveals that the direction of spiralization of helices may be either left or right handed and that the direction of spiralization of a single helix can reverse. The specific location of helices and of 850 Å–1150 Å electron dense bodies suggests that they are either primary chromosome products which subdivide before entering the cytoplasm, or units for the intranuclear transportation of primary chromosome products. In each nuclear membrane pore complex one central and eight peripheral regions of dense material are found. At each of the nine points, the dense material appears to traverse the nuclear membrane.     

Histology and ultrastructure of the neural lobe of the lizard,Klauberina riversiana

Summary The pars nervosa of Klauberina riversiana belongs to a primitive tetrapod type which is characterized by the deep penetration of the infundibular recess, a thin-walled structure, and the virtual absence of pituicytes. The differential response of this gland to aldehyde fuchsin and periodic acid Schiff suggests the presence of two types of neurosecretory nerve endings. Ultrastructurally four kinds of nerve endings are distinguishable. Type I, probably a cholinergic nerve ending, contains only small clear vesicles ca. 400 Å in diameter. The relative abundance of cholinergic nerve endings in this pars nervosa may be related to the necessity of transporting hormone through the ependymal cell. Type II, containing granulated vesicles about 1,000 Å in diameter and probably aminergic, is very rare. The two remaining types apparently secrete neurohypophysial hormones. They are Type III, containing dense granules ca. 1,500 Å in diameter and Type IV containing pale granules ca. 1,500 Å in diameter. Evidence is reviewed which suggests that Type III nerve endings may secrete arginine vasotocin while Type IV endings may secrete (an)other hormone(s).All these axons end only on the ependymal cells, the vascular processes of which form a continuous cuff over the basement membranes of the blood vessels. Hence the ependymal cells link the cerebrospinal fluid, the nerve endings and the blood vessels. Particles resolvable with the electron microscope are traced through a possible transport pathway from the granules, through the ependymal cells to the basement membrane. It is suggested that pituicytes replace ependymal cells and assume their transport functions in animals with massive neural lobes containing large numbers of nerve endings and blood vessels.Fellow of the Consejo Nacional de Investigaciones Científicas y Técnicas de la República Argentina.This investigation was supported in part by a Public Health Service fellowship 1 FZ HD 32,949-01 REP from the national Institute of Child Health and Human Development.The authors wish to thank Professor H. Heller for his constant interest and constructive criticism.     

Ultrastructure of the peptidergic and monoaminergic neurons in the hypothalamic neurosecretory system of Anuran Batracians

Summary In the toad Bufo arenarum Hensel the following regions of the hypothalamic — neurohypophyseal system were studied under the electronmicroscope: preoptic and paraventricular nuclei, median eminence and infundibular process of the neurohypophysis.Neuronal perikarya of the preoptic nucleus are loaded with typical neurosecretory granules of peptidergic nature having a mean diameter of 1660 Å. While most neurons of the winter toad are in a storage stage a few show signs of a more active synthetic activity. A distinctive feature of preoptic neurons is the presence of large lipid droplets. The paraventricular nucleus contains small neurons containing granulated vesicles with a mean diameter of 800-1000 Å. In the region extending between these two nuclei and the median eminence axons containing either neurosecretory elementary granules or granulated vesicles are observed.The inner zone of the median eminence is occupied by axons of the preoptic neurohypophyseal tract; two types of axons, according to the size and density of the neurosecretory granules, may be recognized. The outer zone of the median eminence contains mainly axons and nerve terminals containing granulated vesicles of probable monoaminergic nature and only a few with granules of peptidergic type.The neurohypophysis contains two kinds of axons: one with more dense granules of 1800 Å and the other with granules of lesser electron density and 2100 Å. At the ending proper small clear vesicles of synaptic type are found.A progressive increase in volume of the peptidergic granules along the axon is demonstrated. This is of the order of 218% from the preoptic perikarya down to the infundibular process. The physiological significance of the two neurosecretory systems — i.e. the monoaminergic and the peptidergic — and the probable nature of the two types of peptidergic axons is discussed.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 want to express their gratitude to Mrs. Defilippi-Novoa and Mr. Alberto Sáenz for their skillful assistance.     

Degeneration of monoamine nerves in anterior byssus retractor muscle of Mytilus induced by 5,6-dihydroxytryptamine

Summary Preliminary ultrastructural studies on the effects of 5,6-Dihydroxytryptamine (5,6-DHT) on the anterior byssus retractor muscle (ABRM) of Mytilus show degeneration of 2 types of monoaminergic nerves after 10 days of drug treatment. One type contained large granular vesicles (560–1,680 Å) while the other had small granular vesicles (200–640 Å). These axons may possibly represent serotonergic and dopaminergic nerves, thought to innervate this muscle.Two other types of profiles seemed to be unaffected by the drug. One conforms to cholinergic nerves while the other has a predominance of large opaque vesicles (1,200–2,500 Å). The significance of these findings is discussed in the light of recent observations on the neurotoxic effects of 5,6-DHT on vertebrate and molluscan nerves.The author is grateful to Professor G. Burnstock for research facilities and Professor B. M. Twarog for advice and encouragement. This work was supported by the Ramaciotti Foundation     

Electron microscopic studies on the pars intermedia of the ferret

Summary The structure of the pars intermedia of the ferret has been studied with the electron microscope, with particular reference to the morphology of the glandular cells and their innervation. Two types of cell were found. The predominant cell is ovoid in shape and contains membrane-bound vesicles of varying size (1,000–5,000 Å) and density, the most electron-dense of which are associated with the Golgi region. The nucleus is indented and the cytoplasm contains rough endoplasmic reticulum. The second cell type is often associated with the colloid material and is elongated or stellate-shaped with long processes which extend between the predominant cells. It is devoid of cytoplasmic vesicles and has a poorly defined Golgi apparatus. Certain other structural features of this cell such as microvilli, cilia or cytoplasmic microfilaments are reminiscent of ependymal cells.Numerous nerve endings are observed throughout the pars intermedia, making synaptic contact with the predominant cell type. The majority contain vesicles with an electron-dense core measuring 750 Å; less frequently terminals contain dense granules measuring 1,000 A or more. Both also contain small electron-lucent vesicles (200–400 Å); occasionally terminals containing only the latter type are found. The pattern of innervation in the ferret is thus comparable to that previously observed in the cat, rather than that seen in rodents or monkeys, and the implications of this finding are discussed.We are indebted to Prof. Sir Solly Zuckerman, O. M., K. C. B., F. R. S., for his help and guidance and to Mr. J. Wallington for his unfailing technical assistance.     

On different types of nerve endings in the frog median eminence after fixation with permanganate and glutaraldehyde-osmium

Summary In the frog median eminence, fixed with glutaraldehyde and osmium tetroxide, four types of nerve endings can be generally distinguished. These endings are in contact with the pericapillary spaces of primary portal vessels and can be identified by the internal structure and the size of their granules and vesicles. Type 1 contains large granules (1500–2400 Å in diameter) and small clear vesicles (300–500 Å in diameter), type 2 intermediate granules (about 1100–1700 Å in diameter) and small clear vesicles, type 3 small granules (about 600–1000 Å in diameter) and small clear vesicles, type 4 only numerous small clear vesicles. The mixed types containing the large, intermediate and small dense granules in the same ending are infrequently found.After KMnO₄ or LiMnO₄ fixation the granules and vesicles mentioned above are observed as follows. The large granules in the type 1 nerve ending appear mostly pale or less-dense. The intermediate granules in the type 2 also appear mostly pale or less-dense, but some frequently show granules of high density. The small granules in the type 3 consistently contain the dense substance and these endings can be subdivided into two different types according to the populations of different sizes of dense granules [type 3a (900–1000 Å) and type 3b (500–800 Å)]. Dense-cored and cleared-synaptic vesicles are frequently present with together in the type 3 endings. The small vesicles (300–400 Å), in the type 4, appear generally pale (type 4a), but some nerve endings contain small dense cored-vesicles (type 4b).The author wishes to thank Prof. H. Fujita for his advice and criticism.     

Fine structure of the autonomic nerves of the rabbit myometrium

Summary The fine structure of the preterminal nerve fibers of the rabbit myometrial smooth muscle was studied using potassium permanganate fixation or glutaraldehyde fixation with postosmification. The preterminal fibers were mostly formed by 2–10 axons enveloped by Schwann cells. Two kinds of axons and axon terminals were found. (1) Adrenergic axons, which contained many small, granular vesicles (diameter 300–600 Å) and large granular vesicles (diameter 700–1200 Å) which represented ca. 2% of the total count of the vesicles. (2) Nonadrenergic axons, which contained small agranular vesicles (diameter 300–600 Å) and large granular vesicles (diameter 700–1200 Å). Both types of axons formed preterminal varicosities along their course. The real terminal varicosities, representing the anatomical end of the axons, were usually larger than the preterminal ones and showed close contact to the plasma membranes of the smooth muscle cells. Both adrenergic and nonadrenergic terminals were found close to the smooth muscle cells, but a gap of at least 2000 Å was always present between the two cell membranes. The axons and preterminal varicosities of both types of nerves were in intimate contact with each other within the preterminal nerve fiber. Axo-axonal interactions between the two types of axons are possible in the rabbit myometrium. The relative proportion of the nonadrenergic axons from the total was about one fourth.     

Ciliated neurons and different types of synapses in anterior hypothalamic nuclei of reptiles

Summary The magnocellular paraventricular and supraoptic nuclei and the parvocellular preoptic and periventricular nuclei have been studied by light and electron microscopy in Emys orbicularis, Lacerta agilis and Elaphe longissima. The ultrastructure of cerebrospinal fluid (CSF)-contacting neurons was described in the preoptic and periventricular nuclei of Emys and Lacerta species. Single 9×2+0 cilia similar to those of the CSF-contacting dendritic terminals were found on perikarya of non CSF-contacting nerve cells, in all four investigated nuclei. The cilia project from funnel-like invaginations of the perikarya into the intercellular space. In the neurons of the nuclei studied, granular vesicles were found, their size being mainly 1,600 Å in the paraventricular nucleus, about 1,800 Å in the supraoptic nucleus, 1,100 Å in the periventricular nucleus and 800 Å, or up to 1,250 Å in the preoptic nucleus. In general, the neurons possess synapses of the axo-somatic, axo-somatic spine, axo-dendritic and axo-dendritic spine types. In the supraoptic nucleus, multiple interdigitated synapses were observed. Presynaptically, either synaptic vesicles only, or synaptic vesicles and dense core vesicles of different sizes (600 to 800 Å, about 1,100 Å, 1250 Å, and up to 2,000 Å) were found. It is discussed whether the above described 9×2+0 cilia may represent some kind of hypothalamic sensory structure that earlier physiological studies postulated to exist. The ciliated hypothalamic perikarya are considered by the authors to be a more differentiated form of the CSF-contacting neurons. The different types of synapses indicate multilateral connections of the nerve cells of the nuclei studied.Dedicated to Prof. Dr. Berta Scharrer on the occasion of her 70th birthday     

Vesicle and granule content of sympathetic ganglion cells during limb regeneration of the newt Triturus

Summary Fine structural observations were made on the vesicle and granule content of ganglion cells in the posterior subclavian ganglion and peripheral nerve fibers of the upper forelimb of the newt Triturus. The populations of vesicles and granules in normal ganglion cells and nerve fibers were compared with those observed after limb transection. In normal neurons, clear vesicles range in size from 250 to 1000 Å in diameter, but are most frequently 400–500 Å. Vesicles with dense contents (granules) also vary greatly in size, but most are 450–550 Å in diameter and correspond to dense-core vesicles. Large granules that contain acid phosphatase activity are thought to be lysosomes. During limb regeneration, in both the ganglion cells and peripheral nerves, the ratio of dense vesicles to clear vesicles increases. There is a large increase in number of dense granules with a diameter over 800 Å, particularly in the peripheral regenerating fibers. This study shows that regenerating neurons differ from normal in their content of vesicular structures, especially large, membrane-bounded granules.This work was supported by grants from the National Science Foundation (GB 7912) and from the National Cancer Institute (TICA-5055), National Institutes of Health, United States Public Health Service.     

Electron microscopic observations on synapse-like contacts between pituicytes and different types of nerve fibers in the anuran pars nervosa

Summary Pituicytes of Rana pipiens could be classified into two types, pale and dense, according to their relative densities of cytoplasm and the populations of free ribosomes and cell organelles. An intermediate type of pituicyte was also recognized.Lipid droplet such as are typical in the cytoplasm of mammalian pituicytes, are not in the cytoplasm of either types of frog pituicyte. Both types have long cytoplasmic processes which run among the nerve fibers, and some of them end at the pericapillary space.Nerve endings making synapse-like contacts with the cell bodies or the processes of the pituicyte are frequent. According to the structures and sizes of granules and vesicles in the nerve endings, these endings are classified into one of three types: 1) A, which appears to be a peptidergic neuronal ending containing dense granules 1,200–2,000 Å in diameter and small clear vesicles 300–400 Å in diameter; 2) B, which appear to be monoaminergic endings containing cored vesicles 600–1,000 Å in diameter and small clear vesicles 300–500 Å in diameter; 3) C, which appear to be cholinergic endings containing only small clear vesicles. Type C endings are relatively rare. In the synaptic area the axonal membranes appose those of the pituicytes across a gap of about 200 Å and numerous presynaptic vesicles are clustered or accumulated near the presynaptic membranes.The author wish to express his hearty thanks Professor Dr. A. Gorbman, Zoology Department, University of Washington, Seattle, U.S.A. and Professor Dr. H. Fujita for their helpful advices and criticisms. The frog tissues were obtained and fixed in Professor A. Gorbman's laboratory supported by U.S.P.H.S. grant NS 04887.