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.     

Effect of transection at different levels of hypothalamus on the hypothalamo-hypophysial system of the toad,Bufo bufo,with particular reference to the ultrastructure of the zona externa of the median eminence

Summary The ultrastructural changes taking place in the median eminence of Bufo bufo 3 hours to 4 months after transection of the brain at different levels, are described.5 types of neurons in the zona externa of the median eminence of normal toads are described. All types of neurons degenerate, and profound changes of the ultrastructure of the capillaries are observed after transection just behind, or immediately in front of the optic chiasma. A few neurons containing dense granules with a diameter of about 1,000–1,300 Å remain intact, however. The degeneration following denervation in front of the optic chiasma was considerably delayed compared to degeneration after denervation behind the optic chiasma.After transection more rostral to the optic chiasma, no significant degeneration of the median eminence was observed.The results are discussed with regard to degenerative dynamics and origin of the different nerve types. It is concluded, that all types of neurons terminating in the median eminence, originate at a level between the caudal and rostral parts of the preoptic nucleus, some fibres, however, containing dense, 1,000–1,300 Å granules, originate caudally to the optic chiasma, in the posterior hypothalamus.Part of this study was presented at the Vth International Symposium on Neurosecretion, Kiel, Germany, August 1969.     

The fine structure of the infundibular process of the hedgehog

Summary The fine structure of the infundibular process of the hedgehog has been studied, using material fixed in osmium tetroxide and embedded in Vestopal W. The process resembles in general structure that of other mammals, but also shows features not previously described in other species.The nerve fibres contain a number of inclusions, namely: small vesicles, 300–500 Å in diameter; larger vesicles, up to 2000 Å in diameter, which contain a variable amount of osmiophilic material; hexagonal crystal-like bodies, approximately 1250 × 3000 Å in size, lying within a striated membranous sheath; and aggregate bodies made up of small electron dense granules, possibly derived from mitochondria.In addition complex multilamellate bodies occur in some nerve fibres, which apparently give rise to membranous vesicles. Pituicytes, of varying appearance, are often intimately related to the nerve fibres.The findings suggest that synthesis of material may occur in the distal part of the fibres of the hypothalamo-hypophysial tract.We are indebted to the Medical Research Council, who provided the electron microscope in the Department of Anatomy, University of Birmingham, which was used in this study.     

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.     

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.     

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.     

Ultrastructural study of neurovascular contacts in the median eminence of the rat

Summary In the median eminence of the rat axons of the supraoptic-paraventricular-hypophyseal tract with elementary neurosecretory granules (150–200 m) traverse the internal zone. Terminals containing dense core vesicles 60–120 m in diameter end on the portal capillaries of the median eminence. A unique organisation of the primary portal capillaries is shown. Endothelial cells have many fenestrae. The pericapillary space has numerous extensions all of which represent a special zone around the endothelial tube. The fine structural organisation and function of the neurovascular contacts in the median eminence are discussed.     

Ependymal specializations

Summary The primary plexus of the toad hypothalamic-adenohypophysial portal system has two types of loops. The short loops are localized in the external region of the median eminence and surrounded by nerve endings and glial cells. The long loops approach the ependymal lining of the median eminence. The ascending and descending branches of these loops are surrounded by nerve and ependymal endings and glial cells. The actual subependymal portion of the long loops is virtually in contact with ependymal processes only, which form a cuff interposed between this portion of the long loops and the fibres of the hypothalamic-neurohypophysial tract. Many of the vascular endings of the ependymal processes have electron dense granules whose diameter ranges between 700 and 1400 Å. The ultrastructure of the ependymal cells suggests that these granules are transport material and not secretory material.This anatomical arrangement linking the ependyma of the median eminence and the long loops of the primary plexus of the hypothalamic-adenohypophysial portal system makes the possibility of an interrelationship between the cerebrospinal fluid and the portal blood very considerable.Fellow of the Consejo Nacional de Investigaciones Científicas y Técnicas de la República Argentina. The author takes great pleasure in thanking Prof. H. Heller for his constant interest and criticism.     

Electron microscopic autoradiography on the localization of serotonin in the frog median eminence

Summary For the identification of the nerve fiber containing serotonin (5-hydroxytryptamine) in the frog median eminence, an electron microscopic autoradiography was performed with 5-hydroxytryptophane-³H which is the precursor of serotonin. At 1 and 24 hours after the intraperitoneal injection, most silver grains were located over the nerve fibers and endings, and a few were also found over the glia cell and the perivascular space. A large number of silver grains were located over the type 3 nerve endings (Nakai, 1971) containing small dense granules about 600–1000 Å in diameter 1 and 24 hours after the injection. Some silver grains were localized over the nerve endings containing intermediate-size dense granules 1100–1700 Å in diameter. Silver grains were also frequently observed over the nerve fibers in the inner layer of frog median eminence. There is no significant difference in the pattern of distribution of silver grains between tissues of 1 hour and 24 hours after the injection.The authors wish to thank Prof. H. Fujita for his advice and criticism.     

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.     

Ultrastructural Identification of gastrin-like immunoreactive nerve fibres in the brain of Xenopus laevis by means of colloidal gold or ferritin immunocytochemical methods

Summary By use of an anti-gastrin serum and colloidal gold- or ferritin-labelled sheep anti-rabbit -globulins, nerve fibres and nerve terminals containing a gastrin-like substance were characterized at the ultrastructural level in the median eminence of Xenopus laevis. These immunoreactive fibres contain neurosecretory granules displaying medium to high electron density and a mean diameter of 75 nm. Labelling intensity varies from granule to granule. This is the first demonstration at the ultrastructural level of the precise location of a gastrin-like hormone in the median eminence of a vertebrate.Supported by the D.G.R.S.T., Contrat no 80.7.0242     

Fluoreszenz- und elektronenmikroskopische Untersuchungen am Zwischenhirn-Hypophysensystem von Passer domesticus

Zusammenfassung Fluoreszierende aminerge Nervenfasern bzw. Endigungen und elektronendichte Granula mit einem Durchmesser von 500–1000 Å wurden bei Passer domesticus in den Perikaryen und im Neuropil des Nucleus infundibularis, im Neuropil des Nucleus supraopticus, im rostralen Abschnitt der neurosekretorischen Bahn (Region der Sehnervenkreuzung) und in der retikulären und Palisadenschicht der Eminentia mediana beobachtet. 2000–2500 Å große kontrastreiche Granula finden sich in den Perikaryen des Nucleus supraopticus, im Tractus supraoptico-paraventriculo-hypophyseus und im Hypophysenhinterlappen. Nach Reserpinbehandlung schwanden sowohl die gelbgrüne Fluoreszenz als auch das elektronendichte Material der 1000 Å und z. T. auch der 500–800 Å großen Bläschen. Dieses Ergebnis spricht dafür, daß beide Granulatypen Monoamine enthalten. Der Nucleus infundibularis ist nach unseren Befunden die wichtigste Quelle der etwa 1000 Å großen Granula in der Palisadenschicht der Eminentia mediana.

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Fluorescence- and electronmicroscopic studies on the hypothalamo-hypophysial system of Passer domesticus

Summary In Passer domesticus, fluorescent aminergic fibers and axon terminals and dense-core vesicles of 500 to 1000 Å diameter were found in the perikarya and the neuropile of the infundibular nucleus, in the neuropile of the supraoptic nucleus, in the rostral common neurosecretory pathway (region of the optic chiasma), in the tubero-infundibular tract, and in the reticular and palisade zones of the median eminence. 2000–2500 Å electron-dense vesicles were found in the perikarya of the supraoptic nucleus, in the proximal division of the common neurosecretory pathway and in pars nervosa of the hypophysis. After reserpine treatment, the yellow-green fluorescence as well as the dense material contained in the 1000 Å vesicles and partly also in the 500–800 Å vesicles disappeared. This suggested that the 1000 Å and the 500 Å dense-core vesicles in the hypothalamo-hypophysial system of Passer domesticus contain monoamines and that the infundibular nucleus is an important source of the 1000 Å dense-core vesicles in the palisade layer of the median eminence.

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Herrn Prof. Dr. Dr. E. Horstmann gewidmet.

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Mit dankenswerter Unterstützung durch die Deutsche Forschungsgemeinschaft.

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Stipendiat der Alexander von Humboldt-Stiftung. (On leave from the Department of Veterinary Anatomy, University of Minnesota, St. Paul, Minnesota, U.S.A.).     

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).     

Ultrastructural immunocytochemical localization of neurophysin and vasopressin in the median eminence and posterior pituitary of the guinea pig

Summary With the use of tissue prepared by freeze-substitution and the unlabelled antibody enzyme technique, neurophysin and vasopressin were localized at the ultrastructural level in the posterior pituitary and median eminence of the guinea pig. In the posterior pituitary neurophysin was found in the large neurosecretory granules (1300–1500 Å) of axons, Herring bodies, and nerve terminals. In some of these axons immunoreactive neurophysin was found outside of granules in the axoplasm. By light microscopy neurophysin was found in both the zona interna and zona externa of the median eminence; this was confirmed by electron microscopy. In the zona interna as in the posterior pituitary, neurophysin was localized both inside and outside the large neurosecretory granules. In the zona externa, immunoreactive deposit was primarily located in granules with a diameter of 900–1100 Å in nerve terminals abutting on the primary portal plexus. The distribution of vasopressin paralleled that of neurophysin except that the hormone was rarely extragranular. These results demonstrate for the first time that both neurophysin and vasopressin are present in granules of axons that are in contact with the hypophysial portal vasculature.The authors wish to thank Dr. Alan Robinson for the gifts of antiserum to bovine neurophysin I and for purified bovine neurophysin I; Dr. Ludwig Sternberger for the peroxidase-anti-peroxidase complex; and Dr. Robert Utiger for antiserum to lysine vasopressinSupported in part by U.S. Public Health Service grant RR-00167 to the Wisconsin Regional Primate Research Center from the National Institutes of Health. Primate Center publication No. 14-017.Recipient of NIH, NINDS Teacher-Investigator Award NS-1108.     

Etude au microscope électronique de la neurohypophyse de Rana esculenta L.

Résumé L'éminence médiane et la pars nervosa de Rana esculenta diffèrent du point de vue de leur structure.L'éminence médiane se compose de 2 zones différentes: la zone externe placée près du lobe distal et la zone interne située sous l'épendyme. Dans la zone externe, on distingue, d'après la taille et la forme des grains de sécrétion, 5 types de terminaisons.1. des terminaisons avec de fins granules sphériques denses de 800 à 1000 Å de diamètre; 2. des terminaisons avec des granules de 1000 à 1200 Å de diamètre; 3. des terminaisons avec des grains de forme irrégulière de diamètre supérieur aux précédents (1200 à 1600 Å); 4. des terminaisons avec de volumineux grains denses sphériques d'environ 1200 à 1800 Å de diamètre; 5. un petit nombre de terminaisons ne contenant que des vésicules. Les terminaisons des catégories 3 et 4 sont probablement du type neurosécrétoire.La zone interne contient de nombreuses fibres neurosécrétrices. Elles sont de 2 types, l'une avec de gros granules (1600–2400 Å), l'autre avec des granules moins volumineux (1300–2000 Å). Des fibres non neurosécrétrices ont également été observées.Dans la pars nervosa, on rencontre deux types principaux de fibres neurosécrétrices, l'une avec des grains denses de 1600 à 2400 Å de diamètre, l'autre avec des grains moins denses d'environ 1300 à 2000 Å de diamètre. Dans la zone externe bordant la pars intermedia des fibres aminergiques avec de fines granulations ont été observées.

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Electron microscopic study of the neurohypophysis of Rana esculenta L.

Summary The median eminence and the pars nervosa of Rana esculenta have a different structure.The median eminence has 2 different zones: the outer zone situated near the pars distalis and the inner zone under the ependyme. In the outer zone there are, according to the size and the shape of the granules, 5 types of nerve terminals.1. Endings containing spherical fine dense granules of 800 to 1000 Å in diameter; 2. Endings with spherical granules from 1000 to 1200 Å in diameter; 3. Endings with granules of irregular shape which are bigger than the former (1200 to 1600 Å); 4. Endings with spherical dense granules of about 1200 to 1800 Å in diameter; 5. A few endings containing only clear vesicles. Type 3 and type 4 endings are probably neurosecretory.The inner zone contains numerous neurosecretory fibres. They are of two types: one with big granules (1600–2400 Å), the second with smaller granules (1300–2000 Å). Non-neurosecretory fibres have also been observed.The pars nervosa contains two principal types of neurosecretory fibres: one with dense granules of 1600 to 2400 Å in diameter, the other with lighter granules of about 1300 to 2000 Å. In the external zone lining the pars intermedia, aminergic fibres with fine granules have been observed.

Je tiens à exprimer mes vifs remerciements, à Monsieur le Professeur E. Follenius pour l'intérêt constant qu'il prend à ce travail. Je remercie également Madame R. O. Clauss, collaboratrice technique et Madame Schwoerer, photographe, pour leur aide précieuse.     

Observations on the ultrastructure of nerve cells in the brain of the earthworm,Eisenia foetida,with special reference to neurosecretion

Summary The submicroscopic structure of nerve cells in the brain of the earthworm Eisenia was studied. Six types of neurons containing morphologically different inclusions are identified. Types 1, 2 and 3 contain vesicles filled with homogeneous materials of high electron density. These are essentially similar to elementary granules in neurosecretory systems of vertebrates and invertebrates. Type 4 shows dense-cored vesicles which resemble in size catechol-containing granules as described, for example, in chromaffin cells of the adrenal gland. Type 5 has clear vesicles with a mean diameter of 400 Å. Some of these vesicles have a dense osmium deposit. Type 6 contains electron lucent vesicles with diameters of 500–800 Å. Occasionally these have osmiophilic cores. Clear vesicles of types 5 and 6 are similar to classical synaptic vesicles, while granulated vesicles resemble in size and appearance those described in adrenergic nerve endings. All six vesicle types have the same mode of origin from Golgi membranes. All of these vesicles are considered to be discharged from the perikarya into the axons entering the neuropil.     

The neurohypophysis of the foetal monkey

Summary The neurohypophysis of foetal macaque monkeys has been studied by optical and electron microscopy. Abundant elementary neurosecretory granules are present in the infundibular process by the middle third of gestation. Most of these are of variable electron density, and surrounded by a membrane larger than the granule, so that they appear haloed. A few fibres contain membrane-bounded electron-dense granules which show no halo. Inclusions of synaptic vesicle size are rare. The infundibular stem contains a few fibres with typical inclusions smaller than 1000 Å in diameter; the latter resemble inclusions in fibres of the median eminence.Dedicated to Professor W. Bargmann on his 60th birthday.     

Synaptische Strukturen und Elementargranula in der Neurohypophyse des Meerschweinchens

Zusammenfassung Die Ultrastruktur des Nucleus infundibularis tuberis, der Zona interna, der Zona externa und des Hypophysenhinterlappens von Meerschweinchen wird beschrieben. Vergleichende Beobachtungen an den Nervenfasern der genannten Gebiete zeitigen folgende Ergebnisse:Die Axone der Zona interna und des Hypophysenhinterlappens enthalten gleichgeartete, große neurosekretorische Elementargranula (1,400 Å). Die Axonquerschnitte im Bereich des Nucleus infundibularis tuberis und der Zona externa weisen Granula gleichen Kalibers (800 Å) auf und lassen damit auf einen funktionell-morphologischen Zusammenhang des Nucleus infundiburis und der Zona externa schließen. Ob die kleinen Elementargranula das morphologische Substrat der sogenannten releasing factors darstellen oder ob es sich um adrenerge Axone und Endigungen mit typischen Katecholamingranula handelt, ist nicht zu klären.Es wird unterschieden zwischen echten Synapsen und synapsenähnlichen Kontakten. Nur erstere zeigen alle klassischen Synapsenmerkmale. Ihnen sind im Bereich der Neurohypophyse axosomatische und interaxonale Synapsen zuzuordnen. Zona externa und Hypophysenhinterlappen stellen neurohämale Kontaktzonen dar und zeichnen sich durch gleichartig konstruierte Synapsen aus. Neben interaxonalen Synapsen treten vor allem neurokapilläre Kontakte in den Vordergrund. In der Zona externa sind die Tanyzyten synaptisch mit Axonen verknüpft, so wie im Hypophysenhinterlappen die Pituizyten synaptische Verbindungen mit Nervenfasern eingehen. Die synapsenähnlichen Bildungen im Verlauf und an den Enden der Axone können somit neurokapillärer, neurotanyzytärer und neuropituizytärer Natur sein. Sie enthalten neben den synaptischen Vesikeln auch meist kleine oder große Elementargranula. Die ähnliche Morphologie der synaptischen Bildungen in der Zona externa und im Hinterlappen läßt einen gleichgearteten Inkretionsmechanismus beider neurohypophysärer Abschnitte vermuten. Tanyzyten und Pituizyten könnten dabei eine Überträgerfunktion ausüben. Eine rezeptorische Funktion der Tanyzyten wird gleichfalls diskutiert.Die Drüsenzellen der Pars intermedia der Hypophyse sind durch direkte neuroglanduläre Kontakte mit den Neuronen des Hypothalamus verknüpft. Die Axone stülpen sich hierbei tief in das Zytoplasma der Drüsenzellen ein. Es entstehen dadurch beim Meerschweinchen drei nervöse Endigungsformen, die Synapsen ausbilden: 1. Endigungen, die nur synaptische Vesikel enthalten; 2. Endigungen mit synaptischen Vesikeln und kleinen Granula; 3. Endigungen mit synaptischen Vesikeln und großen Granula.Zwischen den Drüsenzellen der Pars infundibularis der Adenohypophyse sind ebenfalls Axonquerschnitte zu beobachten. Es handelt sich dabei entweder um vegetative Faserbündel oder Nervenfasern mit kleinen Granula wie in der Zona externa (800 Å).

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Summary The ultrastructure of the nucleus infundibularis tuberis, the inner and outer layer of the median eminence and the posterior lobe of the guinea pig are described. Comparative studies on the nerve fibres of these regions lead to the following conclusions:The axons of the inner layer of the median eminence and of the posterior lobe contain similar types of large, neurosecretory elementary granules (Diameter 1400 Å).The cross-sections of the axons in the area of the nucleus infundibularis tuberis and of the outer layer of the median eminence exhibit the same sized granules (Diameter 800 Å), which points to a functional-morphological relationship between the two. Whether the small elementary granules represent the morphological substrate of the so-called releasing factors, or whether it is a question of adrenergic axons and terminations with typical catecholamine granules, is undecided.A distinction is made between true synapses and synapse-like contacts. Only the former display all the classical characteristics of synapses. To this sort must be classed the axosomatic and interaxonal synapses in the neurohypophysis. The external layer of the median eminence and the posterior lobe are distinguished by neurohaemal contact areas and show similarly constructed synapses. Of particular prominence, besides interaxonal synapses, are neurocapillary contiguities. Just as in the posterior lobe the pituicytes show synaptic connections with nerve fibres, so are the tanycytes in the outer layer of the median eminence in synaptical contact with axons. Thus the synapse-like formations along and at the endings of the axons can be of a neurocapillary, neurotanycytial and neuropituicytial nature. They contain, apart from the synaptic vesicles, mainly small but also large elementary granules. The similar morphology of the synaptic formations in the outer layer of the median eminence and the posterior lobe suggests the existence of a similar mechanism of internal secretion in both these neurohypophysial areas. Tanycytes and pituicytes could exercise a carrier function. A possible receptor-function of the tanycytes is considered.The glandular cells of the pars intermedia of the hypophysis are linked by means of direct neuroglandular contacts with the neurons of the hypothalamus, whereby the axons make deep incursions into the cytoplasm of the glandular cells. In the guinea pig three types of termination can be observed that form synapses: 1. terminations that contain only synaptic vesicles; 2. terminations with small granules; 3. terminations with large granules.Cross-sections of axons can also be observed between the glandular cells of the pars infundibularis of the adenohypophysis. Here they can be identified either as vegetative fibre-bundles or as nerve fibres with small granules, such as occur in the outer layer of the median eminence (Diameter 800 Å).

     

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.     

The fine structure of the hypothalamic secretory neurons of the White-crowned Sparrow,Zonotrichia leucophrys gambelii (Passeriformes: Fringillidae)

Summary The fine structures of the neurons and neuropils of the magnocellular supraoptic nucleus and the parvocellular nuclei of the rostral hypothalamus, including the suprachiasmatic and medial, lateral and periventricular preoptic nuclei, and the neuronal apparatus of the organum vasculosum laminae terminalis, have been examined in the male White-crowned Sparrow, Zonotrichia leucophrys gambelii, by correlated light and electron microscopy.The magnocellular supraoptic nucleus is characterized by large neurosecretory perikarya which contain a well developed Golgi complex and densecored granules 1,500–2,200 Å in diameter. The neuropil displays axons, dendrites and glial fibers. Some axonal profiles contain dense-cored vesicles 800–1,000 Å in diameter and clear vesicles 500 Å in diameter. Axo-somatic and axo-dendritic synapses are conspicuous in this nuclear region.The suprachiasmatic nucleus is characterized by an accumulation of small neurons with moderately developed cellular organelles and some dense-cored granules, approximately 1,000 Å in diameter. The profiles of axons within the neuropil contain dense-cored granules 800–1,000 Å in diameter and clear vesicles 500 Å in diameter.The neurons of the medial preoptic nucleus are relatively large and exhibit well developed cellular organelles and dense-cored granules 1,300 to 1,500 Å in diameter. Granular materials are formed within the Golgi complex. The medial preoptic nucleus is rich in secretory perikarya.Occasionally, neurons with granules 1,500–2,200 Å in diameter are encountered in the lateral preoptic and periventricular preoptic nuclei. They may be considered as scattered elements of the magnocellular (supraoptic and paraventricular) system.The organum vasculosum laminae terminalis consists of three layers, i.e., ependymal, internal and external zones, and exhibits a vascular arrangement similar to that of the median eminence. The perikarya of the parvocellular neurons and their axons in the internal zone contain numerous secretory granules ranging from 1,300 to 1,500 Å in diameter.This investigation was supported by Grant No. 5R040 Japan-U.S. Cooperative Science Program of the Japan Society for the Promotion of Science to Professor H. Kobayashi and Professor S.-I. Mikami, by a Scientific Research Grant No. 56019 from the Ministry of Education of Japan to S.-I. Mikami, by support from the Deutsche Forschungsgemeinschaft (Schwerpunktprogramm Biologie der Zeitmessung) to Prof. A. Oksche and by Grant No. GF 33334, U.S.-Japan Cooperative Science Program of the National Science Foundation to Prof. D.S. Farner.Herrn Professor Dr. Dres h.c. Wolfgang Bargmann zu seinem 70. Geburtstag am 27. Januar 1976 gewidmet.