Localization of luteinizing hormone β-mRNA by in situ hybridization in the sheep pars tuberalis

Summary The localization of luteinizing hormone beta (LH)-mRNA was studied by in situ hybridization in the pars tuberalis of sheep using a homologous sheep double-stranded ³²P-or ³⁵S-cDNA. The labelled cDNA probe detected one mRNA sequence in the pars tuberalis by Northern blot analysis; this sequence was similar to that detected in the pituitary. In situ, the labelling of LH-mRNA in the horizontal and sagittal tissue sections was found throughout the pars tuberalis. This labelling was prevented by adding an excess of cold probe or treating the sections by ribonuclease before in situ hybridization. Controls showed a labelling in the pars distalis, but not in the median eminence, hypothalamus, cerebral cortex and liver sections. Double labelling by using a specific LH-antiserum indicated that the labelling of LH-mRNA appeared more intense in LH-containing cells that were found only in the ventral part of the pars tuberalis. These results suggest that the entire pars tuberalis is able to produce the LH subunit, but that the level of translation greatly varies according to the location of the cells.     

Cellular composition of the human pituitary pars tuberalis as revealed by immunocytochemistry

Summary An attempt was made to determine if any of the specialized secretory cell types common to the pars distalis also occur in the pars tuberalis of the human hypophysis. Available for study were 18 specimens of the inferior pars tuberalis, which partially surrounds the infundibular stem, and 3 specimens of the superior pars tuberalis that is attached to the median eminence. Antisera to human somatotropin, mammotropin, chorionic gonadotropin, follicle-stimulating hormone, FSH , luteinizing hormone, LH , thyrotropin, TSH , as well as to ^1–24-corticotropin, porcine ^17–39-corticotropin, and ovine LH were used with the Sternberger peroxidase-antiperoxidase immunocytochemical procedure to identify the probable cells of origin for these hormones.The evidence indicated that gonadotropic cells constitute the major portion of the parenchymal cell population in the pars tuberalis. They occurred throughout all of the pars tuberalis and were usually arranged in clusters. Somatotropic, mammotropic, corticotropic, and thyrotropic cells were rare and not found in all specimens. When present, they often formed a common group suggesting that their occurrence in the pars tuberalis resulted from displacement of primordial tissue of the pars distalis during embryogenesis.Supported in part by research grants HD-03159 and HD-08333 from the National Institute for Child Health and Human DevelopmentWe thank Dr. L.A. Sternberger for providing the PAP complex and others for antisera (Table 2) and hormones (Footnote 2) as listed     

Changes in LHβ-gene and FSHβ-gene expression in the ram pars tuberalis according to season and castration

Luteinizing hormone beta (LH) and follicle stimulating hormone beta (FSH) subunits and their mRNAs were studied in the ram pars tuberalis following different seasonal (winter vs summer) and experimental (intact vs castrated animals) conditions. Hormone-containing cells were identified by immunohistochemistry, and mRNAs for LH and FSH by in situ hybridization using homologous double-stranded ³⁵S-cDNAs. The labelling was quantified by image analysis. Immunohistochemical staining showed that cells containing LH and FSH were localized mainly in the ventral part of the pars tuberalis but that, in the summer, additional LH-containing cells were present in the dorsal part in intact rams. On the other hand, LH-mRNA labelling was found in the whole pars tuberalis in wethers but only in the ventral part in intact rams. The magnitude of LH-mRNA labelling was significantly greater in summer than in winter rams, and in castrated than in intact animals (P<0.001). However, the number of labelled cells was found to be the greatest in the winter (P<0.001) and was not affected by castration. FSH-mRNA expression was similar to that of LH-mRNA except that the level and extent were considerably lower. Thus, our results show an increase in the magnitude of gonadotropin subunit-mRNA in the summer and following castration; this increase appears to involve the entire pars tuberalis.     

Ontogenesis of cells producing polypeptide hormones (ACTH,MSH, LPH,GH, Prolactin) in the fetal hypophysis of the rat: Influence of the hypothalamus

Summary The ontogenesis of cells containing polypeptide hormones (ACTH, MSH, LPH, GH and Prolactin) was investigated in the fetal rat hypophysis by immunohistochemistry using the peroxidase-antiperoxidase complex.Corticotrophs, melanotrophs and lipotropic cells were revealed earlier in the pars distalis than in the pars intermedia. In the pars distalis, cells producing LPH were found in the morning of day 15 of gestation using anti-- or anti--LPH sera, and in the afternoon using anti-- or -endorphin sera. Cells containing -MSH were observed from the afternoon of day 15. The cells stainable with the anti--MSH, anti--(17-39)ACTH and anti--(l-24)ACTH sera appeared on day 16. In the pars intermedia, the cells producing -MSH, MSH, - and -endorphin, and -LPH were observed in the morning of day 17, while cells containing ACTH were only revealed in the afternoon of the same day of gestation. Based on the treatment of serial paraffin sections with various antisera, it was clearly shown that MSH, ACTH, and LPH occur in the same cells located in the pars distalis as in the pars intermedia.The development of the corticotrophs, melanotrophs and lipotropic cells does not require the presence of the fetal hypothalamus or other central nervous structures. The pituitary glands of 21 day-old fetuses encephalectomized on day 16 showed as many reactive cells as those of the littermate controls.The somatotrophs were first revealed in the pars distalis in the afternoon of day 19. The cells producing prolactin were not observed before day 21 of gestation. On some cases GH and prolactin were found together in one cell. The cytodifferentiation of GH and prolactin cells is apparently not under hypothalamic control.     

Identification of the LH and TSH-secreting cells in the pituitary gland of the rhesus monkey

Summary Pituitary glands from juvenile (pre-pubertal) and adult male and female rhesus monkeys were examined following immunocytochemical staining with antisera to the beta subunits of ovine luteinizing hormone (LH) and of human thyroid stimulating hormone (TSH). The LH antiserum reacts with a cell that is PAS-positive, occurs singly and is randomly distributed throughout the pars distalis. The diameter of these cells is approximately 11.5 m. They do not seem to vary in number in either juveniles (pre-pubertals) or adults, or in males or females. There appears to be fewer LH cells in the pituitary glands of pregnant and lactating females. In addition to staining cells in the pars distalis, the antiserum also reacts with a population of cells located in the pars tuberalis.The cells that stain with the anti-TSH serum are confined primarily to the pars distalis. They are approximately 15.8 m in diameter and are generally found in groups or clusters located in the anterior and medial regions of the gland. The TSH cells vary in number from one animal to another; however, this variability is unrelated to the age or the sex of the animals. No demonstrable changes occur in the number of TSH cells during pregnancy or lactation.Supported by NIH General Research Support Grant RR05654The author wishes to express appreciation to the Hormone Distribution Program of NIAMDD for the preparations of ovine FSH, TSH and human TSH, and to Drs. H. Papkoff for the ovine LH and LH, L. Reichert for the human FSH, J. Vaitukaitis for the anti-human TSH, and L.A. Sternberger for the PAP complex     

Immunohistochemical study of the pars tuberalis of the adenohypophysis in the monkey,Macaca irus

Summary The pars tuberalis of the hypophysis in the monkey Macaca irus encompasses the hypophysial stem up to the median eminence. Histologically, it consists of several layers of chromophobic cells. A few PAS¹-positive cells also stainable with Alcian blue (pH 3.0) can be observed among the unstained elements. Using the indirect immunofluorescence antibody technique, scattered immunoreactive cells were revealed with the anti-oLH antibody; these cells did not react with the anti-hFSH antibody. In contrast, the immunoreactions to anti-hGH, anti-hPRL, anti-ACTH, anti-MSH, anti-LPH and anti-endorphin sera were completely negative. Single cells reacting with the anti-hTSH serum were observed at the inferior end of the hypophysial stalk (zona tuberalis), i.e., beyond the pars tuberalis proper. These results are compared with data reported in the literature.

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Résumé La pars tuberalis de l'hypophyse du Singe Macacus irus entoure la tige infundibulaire jusqu'à l'éminence médiane. En techniques histologiques, elle apparaît constituée de plusieurs assises cellulaires d'aspect chromophobe. On y observe quelques cellules PAS-positives réagissant simultanément avec le bleu Alcian (pH3.0). En technique d'immunofluorescence indirecte, des cellules dispersées sont mises en évidence uniquement avec un anticorps anti-oLH; ces cellules ne réagissent pas avec un anticorps anti-hFSH. L'utilisation d'anticorps anti-hGH, anti-hPRL, anti-ACTH, anti-MSH, anti-LPH et antiendorphines ne permet pas de révéler des cellules immunoréactives. Quelques cellules réagissant avec un anticorps anti-hTSH s'observent à la base de la tige hypophysaire (zona tuberalis), c'est-à-dire au-delà de la pars tuberalis proprement dite. Ces résultats sont confrontés à ceux rapportés dans la littérature.

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Abbreviations used in this Article PAS periodic acid Schiff - oLH ovine luteinizing hormone - hFSH human follicle stimulating hormone - hGH human growth hormone - hPRL human prolactin - ACTH corticotropin - MSH melanotropin - LPH lipotropin - hTSH human thyrotropin - BSA and HSA bovine and human serum albumin     

Hypothalamus and cytodifferentiation of the foetal pituitary gland

Summary To investigate whether the hypothalamus is involved in the cytodifferentiation of the anterior pituitary gland, rat foetuses were encephalectomized in utero on day 16 of pregnancy.Pituitary sections from encephalectomized and normal littermate foetuses were studied on day 21 with the immunofluorescence technique using antibodies and -MSH, anti -MSH, anti -(17–39) ACTH and anti -(1–24) ACTH. On day 16, only the anti -MSH revealed a few cells in the pars distalis but not in the pars intermedia. On the other hand, on day 21, the pituitary cells reacting with antibodies anti -MSH, anti -MSH and anti -(17–39) ACTH were as numerous in the encephalectomized foetuses as in the normal littermate foetuses. The cells revealed with the antibody anti -(1–24) ACTH were less numerous and less fluorescent in the pars distalis and intermedia of the hypophysis of the encephalectomized foetuses.On day 21, the adrenals of the encephalectomized foetuses were atrophied in comparison with those of the normal littermate foetuses but they were larger than on day 16.These data suggest that the cytodifferentiation of the corticotroph and melanotroph cells of the hypophysis occurs without the influence of the hypothalamus which is necessary for the normal release of ACTH.     

An immunohistochemical study of epithelial cells in the posterior lobe and pars tuberalis of the human adult pituitary gland

Summary Pituitary glands were examined using reference staining (hematoxylin and eosin, periodic acid-Schiff and alcian blue) and the peroxidase-labeled antibody method, for 1) invading anterior cells in the posterior lobe, 2) intermediate colloid forming follicles, and 3) pars tuberalis cells.The results showed: 1) that the majority of cases possessed invading anterior cells of various amount. Most of these cells were positive for ACTH^1–18, ACTH^17–39 and -MSH. However, on a few occasions, scattered GH, PRL, FSH, FSH, LH and even TSH cells were also present. 2) Colloid forming follicular cells were mostly ACTH cells, but also contained occasional other hormone-secreting cells. Hormone negative cells were correlated with salivary type epithelium. Well established acinic type salivary glands and ciliated epithelium were negative for any hormones immunohistochemically. 3) Pars tuberalis cells were predominantly gonadotrophs but also included TSH and ACTH cells. Some cells appeared to contain both FSH and LH. When these cells underwent squamous metaplasia, they seemed to lose their hormone secreting activity.Part of this study was supported by a Grant-in-Aid for Cancer Research from the Ministry of Education, Science and Culture, Japan     

Role of plastidial acyl-acyl carrier protein: Glycerol 3-phosphate acyltransferase and acyl-acyl carrier protein hydrolase in channelling the acyl flux through the prokaryotic and eukaryotic pathway

Monoclonal antibodies raised against extracts of the rachis abscission zone of Sambucus nigra L. were selected for high reactivity towards abscission-zone proteins. One antibody (YZ1/2.23) has been shown to cross-react, by both indirect and competition enzyme-linked immunosorbent assay and by Western blotting, with a number of plant enzymes including horseradish peroxidase, rice -glucosidase, almond -glucosidase and the lectins from Phaseolus vulgaris and Erythrina cristagalli.The major N-linked oligosaccharide isolated from horseradish peroxidase has the sequence Man 3(Man6)(Xyl2)Man4GlcNAc4(Fuc3) GlcNAc. This oligosaccharide was found to be a potent inhibitor of the binding of YZ1/2.23 to the intact glycoprotein. The common determinant is therefore contained within this structure.Abbreviations ELISA enzyme-linked immunosorbent assay - Fuc fucose - GlcNAc N-acetylglucosamine - HRP horseradish peroxidase - Ig immunoglobulin - Man mannose - SDS-PAGE Sodium dodecyl sulphate-polyacrylamide gel electrophoresis - Xyl xylose     

Cell types in the fetal pars tuberalis of the human adenohypophysis at mid-gestation

Summary The pars tuberalis of the adenohypophysis was investigated in three human fetuses at mid-gestation by electron microscopy or immunohistochemistry. In addition to gonadotrophs and thyrotrophs, identified by immunohistochemistry and ultrastructural morphology, electron microscopy revealed the existence of an additional differentiated cell type closely resembling pars tuberalis-specific cells known from other species. The role of this cell type in the human endocrine regulation remains to be elucidated.     

Immunocytochemical study of Wulzen's cone of the bovine pituitary

Summary The Wulzen's cone of the bovine adenohypophysis presents a variable development and general arrangement. It is joined to the pars intermedia with no intervening connective tissue. It is covered by a single layer of cubical cell epithelium on the side of the hypophysial cleft. Immunofluorescence reveals the presence of different glandular cell types. The most abundant cells are those demonstrated by an anti-oPRL antibody and are either isolated or clustered. Other cells react with anti-hGH, anti-bLH, anti-oLH or anti-hTSH antibodies. Some cells react simultaneously with anti-MSH, anti-^1–24ACTH, anti-^17–39ACTH, anti-LPH and anti-endorphin antibodies. Cell types other than the numerous prolactin cells appear only as isolated elements. We did not observe cells reacting with anti-leu-enkephalin, anti-met-enkephalin or anti-calcitonin antibodies either in the Wulzen's cone or in the pars distalis or pars intermedia.

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Résumé Le cône de Wulzen de l'adénohypophyse bovine présente un développement et une disposition générale variables. Il est accolé à la pars intermedia dont il n'est séparé par aucune cloison conjonctive. Du côté de la fente hypophysaire, il est revêtu par un épithélium simple, cubique. En immunofluorescence, on observe la présence de divers types de cellules glandulaires: les plus abondantes sont des cellules mises en évidence par un anticorps anti-oPRL, isolées ou groupées en amas. D'autres cellules réagissent avec des anticorps anti-hGH, et anti-bLH, anti-oLH ou anti-/mTSH. Quelques cellules réagissent simultanément avec des anticorps anti-MSH, anti-^1–24ACTH, anti-^17–39ACTH, anti-LPH et anti-endorphine. Mises à part les nombreuses cellules à prolactine, les autres types cellulaires apparaissent constamment sous l'aspect d'éléments isolés. Nous n'avons pas observé de cellules réagissant avec des anticorps anti-leu-enképhaline, antiinet-enképhaline ou anti-calcitonine ni dans le cône de Wulzen, ni dans la pars distalis et dans la pars intermedia.

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Abbreviations used in this Article oPRL ovine prolactin - hPRL human prolactin - hGH human growth hormone - bLH bovine luteinizing hormone - oLH ovine luteinizing hormone - pFSH porcine follicle stimulating hormone - hFSH human follicle stimulating hormone - hTSH human thyrotropin - ACTH corticotropin - MSH a melanotropin - MSH melanotropin - LPH lipotropin - PD pars distalis - PI pars intermedia - PN pars nervosa - HC hypophysial cleft     

Immunohistochemical localisation of substances reactive to antisera against α-and β-endorphin and met-enkephalin in the brain of Rana temporaria L.

Summary In the brain of Rana temporaria, two distinct systems reactive with - and -endorphin antisera, respectively, and with a met-enkephalin antiserum, have been detected immunohistochemically.Neurons reacting with - and -endorphin antisera are located (1) in the preoptic nucleus, and (2) in the pars ventralis of the tuber cinereum. Immunoreactive nerve fibres of both groups of perikarya end in the infundibular floor near the capillaries and the preoptico-hypophysial tract. Control reactions have shown that the immunoreactivity is suppressed by the corresponding antigens, but also by -LPH. In view of these results the immunoreactive systems examined correspond to an /-endorphin system or a lipotropinergic system.Neurons reacting with the met-enkephalin antiserum are located in the paraventricular organ. Intense immunofluorescence was observed in the infundibular floor. Controls show that the labelling by met-enkephalin antiserum is exclusively suppressed by met-enkephalin.In the pituitary gland, on the other hand, - and -endorphin antisera reveal: 1) the MSH/ACTH-like cells of the pars intermedia and 2) the ACTH-like cells of the pars distalis.Supported by the D.G.R.S.T., Contrat no 77.7.0648     

Cell type heterogeneity of intermediate filament expression in epithelia of the human pituitary gland

Summary In the present study we have localized immunohistochemically the intermediate filament proteins of the human pituitary gland (adenohypophysis, pars intermedia and pars tuberalis) by an indirect immunoperoxidase technique or by double immunofluorescence methods and analysed the individual cytokeratin polypeptides using two-dimensional gel electrophoresis. We found that the expression of cytokeratins in different epithelial cells of the human anterior pituitary gland was heterogeneous. Whereas the endocrine cells only expressed cytokeratins 8 and 18, the folliculo-stellate cells exhibited a reactivity for cytokeratins 7, 8, 18 and 19 as well as for GFAP and vimentin. The squamous epithelial cells of the pars tuberalis and the Ratke's cysts showed a more complex cytokeratin pattern of both squamous and simple type. Whereas in many cystic epithelial cells including the pseudo-follicles a triple expression of cytokeratin, vimentin and GFAP could be observed, only some basal cells of squamous epithelial nests coexpressed cytokeratin and vimentin. The differences in the intermediate filament protein distribution are discussed in the light of embryological relationships of the different parts of the human pituitary gland.     

Over expression of integrin α 5 β 1 in human hepatocellular carcinoma cell line suppresses cell proliferation in vitro and tumorigenicity in nude mice

Integrin 5 1 and 2 1 are the major integrin receptors in human hepatocytes. However, in human hepatocellular carcinoma cells it was found that the expression of integrin 5 1 was decreased and another integrin 6 1 increased. In this study, the SMMC7721 human hepatocellular carcinoma cells cotransfected or singlely transfected with integrin 5 and/or 1 cDNAs were established, and designated 5 1.6-7721, 5.3-7721, and 1.6-7721 cell lines, respectively. Transfection with cDNAs of integrin 5 and 1 subunits resulted in the overexpression of each integrin and modified biological properties, including a slowed growth rate, changes in the cell cycle from 15.5% of control cells in the G2/M phase to 12.1%, 9.6% and 9.4% in 5.3-7721, 1.6-7721, 5 1.6-7721, respectively, and a decrease in the Cell Mitosis Index from 1.6 in controls to 0.96, 0.95, and 0.72, and 34%, 28% and 52% derived from colony forming ability, respectively. Tumorigenicity was also tested in nude mice with inoculation of cells subcutaneously. Tumor masses growing in nude mice following inoculation with 1.6-7721,and 5 1.6-7721 cells weighed only 52% or 31% those of control cells. These results indicated that deletion or low expression of integrin 5 1 may play an important role in the development of hepatocellular carcinoma. Therefore, induction of expression of the integrin 5 1 in malignant cells could be a potential means of treating hepatocellular carcinoma.     

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.     

PAS-positive cells of the pars intermedia are calcium-sensitive in the goldfish maintained in a hyposmotic milieu

Summary Cytological changes in the pars intermedia of the goldfish were investigated after adding calcium to deionized water (DW). In fish maintained in DW, the PAS-positive cells are highly stimulated in comparison to cells of fish kept in fresh water (FW). In DW supplemented with calcium at the same concentration as in FW (2 mM/l), the hyperactivity of the PAS-positive cells is prevented. When calcium ions are added 60 h before the animals are sacrificed, the PAS positive cells start to show signs of regression and their granules are stored: the release of the granular material appears to be suppressed by calcium. In the goldfish, the PAS-positive cells, homologous to a similar cell type in the eel, react only very weakly with the PAS technique. The name calcium-sensitive cells appears to be more appropriate in the goldfish for this particular cell type, secreting an unknown factor. This factor, different from the prolactin produced in the rostral pars distalis of the hypophysis, might be an equivalent of a hypercalcin.     

Ultrastructure of the pituitary gland (pars distalis) in sockeye salmon (Oncorhynchus nerka) during gonad maturation

Summary The fine structure of the various hormone-producing cell types (with the exclusion of the prolactin cells) in the pituitary gland (pars distalis) of migratory sockeye salmon is described. All fish were in an advanced stage of sexual maturation. In the proximal pars distalis five cell types were distinguished: growth hormone cells, ACTH cells, gonadotrops, vesicular cells, and chromophobe cells. Gonadotrops were also found throughout the rostral pars distalis. A conspicuous feature of the gonadotrops was the presence of two kinds of secretory inclusions: small electron-dense granules (200–375 m) and large, relatively electron-translucent globules (400–2 000 m). The large vesicular cells, so called because of their conspicuous vesicular endoplasmic reticulum, were numerous and often appeared to contain some small granules. It is argued that they may represent a second type of gonadotropic cell, which, in earlier stages of gonad development, contains many granules but becomes largely degranulated near the time of reproduction when the other gonadotrops (globular gonadotrops) abound. The chromophobes, which were smaller and far less abundant than the vesicular cells, also appeared to contain small granules (120–280 m). They are probably thyrotrops.The assistance of Mr. S. Killick, of the International Pacific Salmon Fisheries Commission, who helped in the collection of salmon, is gratefully acknowledged.     

Plant regeneration via somatic embryogenesis in Styrian pumpkin: cytological and biochemical investigations

Somatic embryo formation was induced from cotyledon explants of Styrian pumpkin (Cucurbita pepo L. subsp. pepo var. styriaca Greb.) by using a solid MS medium supplemented with 16.11M NAA and 4.44M BA or 26.85M NAA and 13.32M BA. The callus proliferation was more efficient on medium supplemented with 26.85M NAA and 13.32M BA. In contrast, the embryogenic response was higher on medium with lower concentrations of growth regulators (16.11M NAA and 4.44M BA). The time needed for embryo induction did not depend on medium composition. Embryos in globular stage were transferred to three different maturation media, containing 2.89M GA₃ in combination with 0.54M NAA, 11.42M IAA and growth regulator-free medium. The germination rate was the highest when embryos were cultured on medium with 11.42M IAA. Plantlets grown on this medium achieved maturity suitable for transplantation into soil within 9 to 10weeks. The regenerated plants were successfully transferred into field and developed fertile flowers and set fruits. Biochemical analysis showed significant lower total glutathione levels among in vitro grown plantlets compared to seedlings grown in soil. When the plantlets were transferred into soil, they reached a normal size within a month and the glutathione concentration was comparable to seed-derived plants at the same developmental stage. Transmission electron microscopy was used to investigate possible differences in the ultrastructure of cells from callus cultures, and leaf cells of regenerated and seed-derived plants. Differences in the ultrastructure were found within chloroplasts which contained only single thylakoids, large starch grains and small plastoglobuli in callus cells in comparison to leaf cells, which possessed a well developed thylakoid system, small starch grains and large plastoglobuli.     

An unusual cell in the central nervous system of Hirudo medicinalis L.: a neuron with ribbons and flags

Summary A neuron (cell 151) with the ability to silence efferent activity in the roots of a leech segmental ganglion was filled with horseradish peroxidase and studied by light-and electron microscopy. The neurites of cell 151 penetrate all areas of the neuropile except for a thin ventral layer. The branching pattern of the secondary neurites is highly variable. Post-and presynaptic structures of chemical synapses with clear vesicles of 25 nm diameter were identified. Neurites are mostly wrapped in glia and run in bundles among other axons. They frequently form ribbons which are 20–40 nm thick, extend several microns away from the dendrite and are followed up to 3 m in depth. They also form flags which are 0.5 m thick, spread out 10–20 m horizontally and run up to 80 m laterally. Both structures lie adjacent to or wrap around axons of other neurons, forming a gap of 8–10 nm. Flags and ribbons are typical for glia but have not been described previously as structures of neurons. Contralateral cells 151 appose each other in the commissures with a gap of 5–10 nm. The possible functional significance of these findings is discussed with respect to electrical coupling and to reception of strain.     

Morphology of horseradish peroxidase (HRP)-injected glial cells in the myenteric plexus of the guinea-pig

Glial cells of the myenteric plexus from guinea pig small intestine were intracellulary filled with horseradish peroxidase (HRP), and histochemically stained. Camera lucida-like drawings of twenty cells were morphologically and morphometrically analyzed. The cells have very small ellipsoid, somata (85±0.7 m equivalent diameter, i.e., about 330 m³ volume), and send up to 20 thin and short processes (less than 26 to about 110 m in length). The morphology of the cells appears to depend on their location within the plexus. Glial cells located within the ganglia are similar to CNS protoplasmic astrocytes; they are star-shaped, and their very short processes are irregularly, branched. In contrast, glial cells within the interganglionic fiber tracts resemble CNS fibrous astrocytes. They extend longer processes that are parallel to the fiber tracts, and show less tendency to branch. We propose that the morphology of enteric glia is determined by the structure of the microenvironment. Both cell types form several flat endfeet at a basal lamina either surrounding blood vessels or at the ganglionic border. Furthermore, the occurrence of holes in the glial cell processes suggests that particular neuronal cell processes may be enwrapped in a specific manner. Fractal analysis of camera lucida-like drawings of the cells showed that the cells have a highly complex surface structure, comparable to that of protoplasmic astrocytes in the brain. These tiny cells may possess a membrane surface area of 2000 m², almost 90% of which are contributed by the cell processes. This geometry may enable an intense exchange of metabolites and ions between neurons, glial cells, and the capillaries and/or environment of enteric ganglia.