On the innervation of the salt gland

Summary The innervation of the salt gland of the goose, the duck and the swan was investigated by means of electron microscopy. Axonal swellings were observed in relationship to secretory cells as well as to central duct cells. The terminals contain synaptic and densecored vesicles. There are no specialized pre- and postsynaptic membranes.Supported by the Deutsche Forschungsgemeinschaft (Ku 210/4).     

Development of the innervation in the human pineal organ

Summary This investigation is concerned with pineal organs of human embryos 60 to 150 days old. At every stage central nerve fibres enter the pineal organ by way of the habenular commissure, but are restricted to the pineal's proximal part. On about the 60th day of the development the sympathetic nervus conarii grows into the distal pole of the pineal organ from a dorso-caudal direction and plays the predominant part in the innervation of the pineal organ. After penetrating, it soon branches out and forms a network in the pineal tissue. Much later, not until the 5th embryonic month, sympathetic nerves appear accompanying the supplying vessels in the perivascular spaces. After a short time these nerves pierce the outer limiting basement membrane and penetrate the parenchyma. Towards the end of the 5th embryonic month the axons of the sympathetic nerves form varicosities containing clear and dense core vesicles. At this point large amounts of laminated granules appear primarily in cell processes, probably of pinealocytes. Isolated granules also occur in the varicosities of axons. The granules encountered here are most likely secretory granules.Dedicated to Professor Bargmann on his 65th birthday.     

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     

The innervation of the salivary gland of the moth,Manduca sexta

Summary The salivary glands of the moth, Manduca sexta (Insecta: Sphingidae), are unlike most other salivary glands in that they are innervated from one source only. Vital staining of nerves with methylene-blue reveals numerous fine nerves extending to the glands from the oesophageal nerve, a part of the stomatogastric or visceral nervous system. Light and electron microscopy confirm that only the fluid-secreting cells, confined to a discrete region in these glands, are innervated. Axons with or without glial wrappings are found in intercellular spaces between fluid-secreting cells. Axons lacking a glial sheath contain, after glutaraldehyde-osmium tetroxide fixation, large granular and small agranular vesicles. In nerve endings in glands fixed with permanganate these smaller vesicles are granular, having the electron-dense cores characteristic of monoamine-containing neurons. These nerve endings with synaptoid areas are in close (direct) contact with the fluid-secreting cells.I am grateful to Professor T. Weis-Fogh for accommodation in the department of Zoology and to Dr. Nancy Lane for use of A.R.C. facilities and advice. Thanks are also due to Drs. M. J. Berridge, S.H.P. Maddrell, and W. T. Prince and Mr. R. A. Leslie for helpful discussion. Financial assistance from Clare College, Cambridge is gratefully acknowledged.     

The structure and innervation of the sinu-atrial node of the mole heart

Summary The sinu-atrial node in the heart of the mole, Mogera wogera, contains myocytes which are devoid of atrial specific granules and which may be classified into two types: electron-lucent (majority) and dark (minority). Numerous unmyelinated axon terminals, containing synaptic vesicles, face the nodal myocyte surface with interspaces of less than 300 nm. There are about 6 terminals per myocyte profile and about 10 per profile at nuclear levels. Of a total of 2717 of these terminals 85.2% are 50–200 nm from the nodal myocytes, and 9.6% form closer neuromuscular junctions, with less than 50 nm interspace and some membrane specialization. Such specializations are almost exclusively found in relation to the dark, minority-type of nodal myocyte.Myelinated nerve fibers are also numerous within the mole SA node, and may terminate almost directly on a nodal myocyte. A few nerve fibers contain many mitochondria and may represent afferent endings.Nodal capillaries are smaller and fewer than capillaries in the juxtanodal myocardium, probably indicating a lesser blood supply to the SA node than to the ordinary myocardium.The author wishes to thank Professor A. Yamauchi, Department of Anatomy, for suggesting this investigation and for constant guidance in the course of the work, Professor K. Shiraishi, Department of Biology, for his encouragement and support, and Miss. K. Ito for technical assistance     

Changes in the surface morphology of the rabbit endometrium related to the estrous and progestational stages of the reproductive cycle

Summary Changes occurring on the surface of the uterine luminal epithelium of the rabbit during the estrous and progestational stages of the reproductive cycle were examined by scanning and transmission electron microscopy. The findings demonstrate that the uterine epithelium, or endometrium, contains two cell types: ciliated cells and nonciliated, microvillous cells. In estrous animals, ciliated cells, although not very numerous, were usually observed in small groups. However, at increasing intervals of time following mating, ciliated cells progressively disappeared from the endometrium until approximately eight to ten days post coitum, when they became scare. From several hours to four to five days following mating, extensive changes occurred on the surfaces of microvillous cells. When observed by TEM, these elements contained organelles typical of cells involved in the synthesis and secretion of glycoproteins. Furthermore, microvillous cells during this period displayed numerous apical protrusions of different sizes and shapes and containing material of varying electron density. Parallel SEM examinations of the same material confirmed the presence of these protrusions. Some of the protrusions appeared as spheroidal masses attached to the cytoplasm by means of a cytoplasmic strand. Other surface masses were clearly unattached to the cell surface and were distributed (1) on the surface of microvillous cells, (2) on the cilia of adjacent ciliated cells, and (3) on the surface of spermatozoa.Changes occurring on the luminal surface during the early postcoital period are interpreted as an expression of morphodynamic processes likely involving coupled secretion (exocytosis) and resorption (endocytosis) of luminal material. The observations presented here also demonstrate that between six and ten days post coitum, the rabbit endometrium contained increasing numbers of enlarged, nonciliated cells that probably arose by the fusion of smaller, microvillous elements.The work reported here was supported by C.N.R. contracts No. CT 760128809 and CT 77014239 (to P.M.) and NIH. Grant HD-04274 (to J.V.B.)     

Phagocytotic and iron-storing capacities of stromal cells in the rat endometrium

Summary Cytoplasmic pigment inclusions of rat endometrial stromal cells were studied by histology, histochemistry, fluorescence microscopy, electron microscopy and X-ray microprobe analysis. It is shown that a number of endometrial perivascular stromal cells contain numerous free cytoplasmic ferritin particles as well as hemosiderin vacuoles. The larger pigment inclusions reveal also positive PAS- and Schmorl reactions indicating that they contain polysaccharide and lipofuscin material, respectively. These pigmentstoring stromal cells also display acid phosphatase activity; they avidly phagocytose instillated latex particles. No pigment-storing cells occur within the surface or glandular epithelium, either in the basal endometrium or in the myometrium.It is demonstrated that the endometrial iron-storing cells function as iron depots; they take part in the phagocytosis and endocytosis of extracellular tissue components and therefore can be named phagocytes. Our data show that fibroblastoid endometrial stromal cells may differentiate into endometrial resident phagocytes, which ensure interstitial proteolysis and hence facilitate the drainage of extracellular fluid into the venous blood capillaries.Supported by a grant from the Nationaal Fonds voor Wetenschappelijk Onderzoek — Fonds voor Geneeskundig Wetenschappelijk Onderzoek, Belgium.     

Quantitative electron microscopic studies on the innervation of the human thymus

Summary Quantitative electron microscopic studies have been carried out on the human thymus. According to the equation L _v =(2n)/F (Hennig, 1963) we have calculated that there is less than 0.204 mm nerve per 1 mm³ thymus tissue inside the blood-thymus-barrier (level of significance of 0.95). This result is compared to the degree of innervation in brown adipose tissue, which contains more than 160 mm nerve per 1 mm³ tissue. The biological significance of the paucity of neuronal elements in the thymus is undetermined.We are much obliged to Dipl. Ing. Dr. A. Hennig for his advice in the mathematical evaluation of our results.We are also indebted to Dr. med. A. Krug (Chir. Universitätsklinik Kiel) for human thymus material.This investigation was supported by the Deutsche Forschungsgemeinschaft.     

The effect of decentralisation or chronic hypogastric nerve stimulation in vivo on the innervation and responses of the guinea-pig vas deferens

Summary The physiological, pharmacological and morphological characteristics of guinea-pig vas deferens supplied by hypogastric nerves rendered inactive by decentralisation were compared with those of vas deferens in which the nerve supply had been chronically stimulated for 3–9 days using implanted electrodes. No change was seen in decentralised preparations prior to 7 days, but from 8–15 days, increased sensitivity to application of noradrenaline in vitro was observed, which was shown to be related to reduced transmitter uptake by nerve terminals as well as to an increase in postjunctional sensitivity; there was also increased fatigability 7–14 days following decentralisation. Continuous stimulation of hypogastric nerves at 2 Hz for 4–8 h daily for 4–8 days resulted in enhanced transmitter uptake and reduced responses to noradrenaline; this was associated with a slight increase in noradrenaline content and a faster adrenergic neuromuscular response with a shorter latency. No appreciable changes in nerve or muscle structure studied by electron microscopy were observed following decentralisation, but there was an increase of between 12.5 and 29.6% in the number of close (< 100 nm) neuromuscular junctions following chronic stimulation for 8 days.     

Observations on the fine structure,enzyme histochemistry,and innervation of parathyroid gland and ultimobranchial body of Chthonerpeton indistinctum (Gymnophiona,Amphibia)

Summary Fine structural and enzyme histochemical observations on ultimobranchial body and parathyroid gland of the caecilian Chthonerpeton are presented. The cell clusters and follicles of the ultimobranchial body consist mainly of granulated cells which are termed C-cells and obviously belong to the APUD cell series. In the larger follicles additional possibly exhausted degranulated cells and replacement cells occur. A rich supply of nerve fibres has been found in this gland. Frequently nerve terminals were observed to come into synaptic contact with the C-cells. Two categories of nerve fibres occur: a) fibres containing large polymorphic electron dense granules (probably purinergic fibres), b) fibres containing small electron transparent vesicles and a few electron dense granules (probably cholinergic fibres). The parathyroid gland consists of elongated cells (one cell type) poor in organelles and often containing fields of glycogen and lipid droplets. The cells are further characterized by fair amounts of lysosomal enzymes; they are interconnected by maculae adhaerentes and occludentes. No nerves and blood vessels have been found in the parathyroid gland of Chthonerpeton. This study has been supported by the Deutsche Forschungsgemeinschaft We 380/5.     

The fine structure of the pancreatic nerves of the domestic fowl

Summary The fine structure of the pancreatic nerves of the domestic fowl has been studied. Naked axon beadings were found in membranous contact with endocrine as well as exocrine cells. From an anatomical point of view it seems reasonable to suggest that the endocrine glands might be subjected to some influence of the autonomic nervous system.     

Innervation of skin glands in the frog

Summary A comparative study was undertaken on the innervation of mucous and granular glands in frog skin. Results obtained by the Falck-Hillarp fluorescence technique and cholinesterase staining indicated that both types of glands receive exclusively adrenergic innervation. Electron microscopy was used to investigate the innervation pattern at the ultrastructural level. The distribution of nerve terminals was found to differ in the two types of glands. In the mucous gland, terminals were found at a distance of about 0.5 m from the basement membrane but never within the gland parenchyma. In the granular gland, the terminals were located between smooth muscle cells and also in direct contact with the secretory epithelium but never outside the basement membrane.This work was carried out in part at King Gustaf V:s Forskningsinstitut, Stockholm, and was supported by a grant from Karolinska Institutet     

On the innervation of mammalian endocrine glands (anterior pituitary and parathyroids)

Summary Electron microscopic studies have been carried out on the innervation of the mammalian anterior pituitary and parathyroids. The total area of grid squares (2.25·10^–2mm²) examined was 2000 per gland and species. In the pituitary pars distalis and in the parenchyma of the parathyroid gland we did not observe a single axon profile. According to the equation proposed by Hennig (1963) we have calculated that there might be—if any—0.133 mm of nerves per 1 mm³ tissue in those two endocrine glands (level of significance 0.95). Comparing these results to the degree of innervation in brown adipose tissue containing more than 160 mm nerve per 1 mm³ tissue we can not imagine that such a small degree of innervation is of any biological importance.In the pituitary pars tuberalis two types of axon terminals have been found both inside and outside the basement membrane surrounding the epithelial complexes. One type contains synaptic and two populations of smaller dense-cored vesicles, the other one contains a population of larger granules which have some properties of the classical elementary granules. Further investigations have to clarify the functional significance of those nerve endings.This investigation was supported by the Deutsche Forschungsgemeinschaft.     

Electron-microscopic investigation of the innervation of the pars distalis of the hypophysis in adult Rana temporaria L.

Summary In the pars distalis of the hypophysis of adult Rana temporaria, three types of nerve-fiber profiles were found at two distinct sites, in both lateral parts of the bordering regions of the anterior lobe with the intermediate lobe of the hypophysis. The first type of nerve-fiber profile consists of bundles of very fine axonal elements (diameter: <0.7 m). The second type is formed by larger nerve fibers (diameter up to 4 m) containing a few neurosecretory granules of approximately 100 nm. The third type of nervefiber profile resembles the second type but these nerve fibers make synaptoid contacts on at least two different types of glandular cells. The possible functional significance of these nerve fibers in the pars distalis is discussed.No nerve fibers were found (1) in the central part of the bordering region of the pars distalis with the intermediate lobe, (2) at the bordering region with the median eminence and (3) with the neurohypophysial stalk, and (4) in all other parts of the pars distalis.     

Innervation of the testicular interstitial tissue in reptiles

Summary In the tortoise Testudo graeca, the lizards Lacerta dugesi and Lacerta pityusensis, and the snake Natrix natrix, the innervation of the testicular interstitial tissue was studied by light and electron microscopy, the acetylcholinesterase (ache) technique, the Falck-Hillarp method for the detection of catecholamines, and the application of 6-hydroxydopamine. The intertubular spaces of the reptilian testes studied contain adrenergic nerve fibers the amount and distribution of which varies considerably both in various species and in various stages of the reproduction cycle. Nerve fibers do not enter the seminiferous epithelium. Fluorescence microscopy of the lizard testis reveals catecholaminergic varicosities which are mainly arranged around blood vessels, but do not show obvious connexions to Leydig cells. Ache-positive fibers are equally distributed in lizard testes surrounding each seminiferous tubule. In Natrix natrix ache-positive fibers are irregularly spread among groups of tubules, without showing a definite relation to Leydig cells either. By electron microscopy bundles of unmyelinated axons and axon terminals can be more easily detected in the testes of immature animals than in adult. Terminals of nerve fibers containing small (400–500 Å in diameter) and large (800–1400 Å) dense-cored vesicles and sometimes small clear vesicles establish contacts with Leydig cells. Three types of contact are described. 1. Contacts par distance at a distance of about 2000 Å and basal lamina interposed; 2. membranous contacts having a 200 Å gap only between axolemma and Leydig cell plasmalemma; 3. invaginations of terminals into Leydig cell perikarya. The latter may exhibit surface specialisations, which strongly resemble postsynaptic membrane thickenings. Experiments using 6-hydroxydopamine underline the adrenergic character of testicular nerve fibers, which can be regarded as another example of non-cholinergic, ache-positive neurons. In the testis of the immature tortoise profiles of axons occur which probably represent purinergic, ache-positive neurons.Supported by a grant from the Deutsche Forschungsgemeinschaft (Un 34/1).I am much indebted to Mrs. R. Sprang for her skillfull technical assistance.     

Development of the innervation of fetal mesenteric microvasculature

Summary The innervation of the mesenteric microvasculature was studied in fetal and neonatal rabbits with the aid of methods demonstrating fluorescence of catecholamines and cholinesterase activity as well as a silver impregnation procedure. The results showed that: (1) adrenergic nerve fibers were present, coursing independently in the mesentery by day twenty-one of gestation, and were found routinely in the adventitia of arterioles and venules by day 25 of gestation; (2) cholinesterase positive cells and fibers of the myenteric plexus were present by day 18 of gestation but cholinergic fibers were not present in the mesentery until day 26; the latter not being associated with blood vessels; and (3) nerve fibers in the mesentery thought to be sensory stained positively with the Holmes silver method on day 18 of gestation.Supported by grants from the Akron Heart Association and the Heart Association of Southwestern Ohio.Recipient of N.I.H. Research Career Development Award AM-42, 370.     

Electron microscopic studies of the innervation of the human spleen

Summary The innervation of four normal human spleens was investigated by electron microscopy. Unmyelinated nerve fibers accompanied the arterial vascular system up to the arterioles of the red pulp. Neither myelinated nerve fibers nor ganglion cells were seen in the splenic hilum or in the splenic tissue itself. The nerve fibers terminated against the smooth muscle cells of the blood vessels in a manner that is typical of the autonomic nervous system. The terminal axons contained small and large granular vesicles and thus were adrenergic nerve fibers. In contrast to the results of previous studies using silver impregnation methods innervation of the red or white pulp could not be demonstrated. The findings on human spleens agree with those on mammalian spleens obtained by other authors using ultrastructural and fluorescence histochemical methods.We are indebted to Prof. Dr. K. Unsicker for his help in discussing the results     

Innervation of the umbilical vessels

Summary Umbilical vessels of guinea-pig fetuses were studied shortly before birth. In all umbilical cords investigated an innervation of the umbilical vessels is lacking. The intrafetal parts of the umbilical vessels on the other hand are richly innervated. A marked difference in the amount of nerve fibres and the pattern of innervation is found between artery and vein. The artery is supplied by a dense nerve plexus which spins around the media and which originates from nerve bundles within the outer adventitial layers. The comparatively scanty innervation of the vein exhibits a more coarsely meshed net pattern. The nerve bundles in the vein exhibit a close affinity to the vasa vasorum.Number and type of the close contacts between the muscle cells are different in the various sections of the umbilical vessels. Similar to the distribution of nerves they are almost absent in the vessels of the umbilical cord, numerously, however, in the intrafetal parts. Contrary to the innervation, the close contacts in the vein are developed more numerously and more broadly than in the corresponding artery.