Influence of Medium Concentration on Prolactin and Growth Hormone Cells During Short-Term Incubation of Pituitary Glands from Tilapia mossambica

Pituitaries removed from Tilapia mossambica adapted either to sea water or fresh water were incubated for 3 or 6 hours in isosmotic or hyposmotic medium. Activation of prolactin cells from pituitaries incubated in hyposmotic medium was indicated by frequent exocytosis and by proliferation of cellular organelles. These pituitaries also showed a reduction in prolactin content and an increase in the amount of bioassayable prolactin in the medium. Incorporation of tritiated leucine into prolactin was least in prolactin cells from seawater Tilapia incubated in isosmotic medium and greatest in those of freshwater Tilapia incubated in hyposmotic medium. These observations indicate that in Tilapia, both synthesis and secretion of prolactin may be stimulated directly by dilution of the medium bathing the pituitary. On the other hand, there were no ultrastructural changes in the presumptive growth hormone cells incubated for 3 or 6 hours in the hyposmotic medium. Hence, these cells do not appear to play a major osmoregulatory role.     

Prolactin regulation in the coho salmon,Oncorhynchus kisutch

Summary Parr and smolt sea water acclimated coho salmon,Oncorhynchus kisutch were subjected to gradual and direct transfers to fresh water. Plasma osmotic pressure, Na⁺, K⁺, Ca⁺⁺ and Mg⁺⁺ were similar in freshwater (FW) fish and seawater (SW) transferred controls for the 24 h following transfer. In spite of the similarity in osmotic pressure and ion levels, plasma cortisol concentrations were significantly increased immediately following salinity change while both pituitary and plasma prolactin decreased indicating enhanced secretion by the pituitary and clearance from the blood. In vitro experiments showed greater incorporation of tritiated leucine into prolactin (PRL) cells immediately after transfer to FW while prolactin injections into intact fish lowered activity in rostral pars distalis (RPD) cells as measured by the same technique, providing evidence of hormonal feedback. These experiments show that the increased synthesis and release of PRL that occurs in coho following movement into FW is not obviously correlated with plasma osmotic pressure, Na⁺ or Ca⁺⁺ concentrations as has been observed in other species of teleosts.Abbreviations FW freshwater - SW seawater - PRL prolactin - RPD rostral pars distalis     

The Response of Prolactin,ACTH, and Growth Hormone Cells in the Pituitary Gland of the Three-Spined Stickleback,: Gasterosteus aculeatus L. form leiurus,to Increased Environmental Salinities

Benjamin, M. 1980. The response of prolactin, ACTH, and growth hormone cells in the pituitary gland of the three-spined stickleback, Gasterosteus aculeatus L. form leiurus, to increased environmental salinities. (Department of Anatomy, University College, Cardiff, Wales, U.K.) — Acta zool. (Stockh.) 61(1): 1–7. The time-sequence of response of the prolactin, ACTH and growth hormone cells in the pituitary gland of the leiurus form of the three-spined stickleback, Gasterosteus aculeatus L., to a transfer from freshwater to seawater, was studied by light microscopy. The appearance of the pituitary was compared with that of animals caught in brackish or sea water. The prolactin cells respond only slowly to seawater by cytological changes interpreted as signs of decreased secretory activity. It is thus suggested that prolactin is unlikely to be important for osmoregulation in this stickleback. The growth hormone cells are more active in all seawater-adapted animals, while the ACTH cells are less active—although only in laboratory experiments. The differing responses of the pituitaries of the leiurus and trachurus forms of G. aculeatus and Pungitius pungitius to high salinities are compared. Even in species whose pituitaries are virtually identical, the response to high salinites may differ widely.     

Ultrastructural morphometric analysis of ameloblasts exposed to fluoride during tooth development

Since a considerable amount of the world population is exposed to high doses of fluoride, it is of special concern to investigate its action mechanisms during dental enamel development. In this study, the toxicity of fluoride in ameloblasts during enamel development was evaluated by means of ultrastructural morphometric analysis. A total of 18 male Wistar rats were distributed into three groups. In Group I, the animals received deionized drinking water ad libitum (negative control) and in Groups II and III, they received sodium fluorided (NaF) drinking water at doses of 7 and 100 ppm ad libitum, respectively, for 6 weeks. Morphometric data were expressed as volume density of the most significant organelles present in the secretory and maturation phases of amelogenesis such as RER, granules, lysosomes, phagic vacuoles, microfilaments and mitochondria. The results showed that the volume density of mitochondria in the 100 ppm experimental group was 29% (P < 0.05) higher than the control group in secretory ameloblasts. No remarkable differences were found in maturation ameloblasts for all organelles evaluated. Taken together, these data indicate that NaF at high doses is able to induce cellular damage in secretory ameloblasts, whereas no noxious effect was observed during maturation stage of amelogenesis as depicted by ultrastructural analysis.     

The occurrence of argyrophilic and argentaffin cells in the gut of Ciona intestinalis L.

Summary Argyrophilic and argentaffin cells occur in the stomach and intestinal epithelium of the sea-squirt, Ciona intestinalis L.. These cells are characterized by their basal swelling which contains the nucleus surrounded by small secretory granules and by a filamentous cell-apex which reaches the gut lumen. The cells are scattered unevenly within the epithelium. Their number decreases rapidly towards the lower part of the intestine. The localization, size of granules and their shape are features which differentiate these cells from other secretory cells in the gut epithelium such as mucous cells. These cells are thought to possess an endocrine function.The excellent technical assistance of Mrs. R. Sprang is gratefully acknowledged     

Ultrastructural studies on prolactin and growth hormone cells in Anguilla pituitaries in long term cultures

Summary Eel hemi-pituitaries were cultured in vitro on high or low sodium media, previously shown to affect differentially prolactin and growth hormone release. After 6 days culture, there were marked differences in the ultrastructure of both prolactin and growth hormone cells from the two groups. Morphometric data on the prolactin cells from SW-adapted eels showed a greater abundance of RER and paucity of secretory granules in cells from the low sodium medium. The size of the Golgi apparatus and the number of exocytosed secretory granules did not differ markedly between experimental groups, in contrast to previous findings on short-term cultures. Differences in the profile diameters of secretory granules are recorded between the experimental groups and the pattern differs markedly from that previously recorded for short-term cultures. The growth hormone cells from low sodium media were characterised by abundant, vesiculated RER, a prominent Golgi apparatus (in SW-adapted animals) and relatively few secretory granules. The activity of these growth hormone cells is in marked contrast to previous findings relating to short-term cultures. The shape and size of the non-granulated (stellate) cells of the RPD was again affected by the osmotic pressure of the medium.I should like to thank Mr. P.F. Hire for his photographic assistance     

Development,structure, and occurrence of secretory trichomes ofPharbitis

Summary Secretory trichomes develop from epidermal cells on the leaf primordia and stem ofPharbitis nil. Following an initial growth phase, trichomes begin active secretion of a protein-carbohydrate mucilage. This mucilage covers the shoot apex and developing leaves ofPharbitis.The secretory cells possess cellular organelles in forms usually associated with actively secreting cells: many mitochondria, an elaborate network of rough endoplasmic reticulum (RER), many free ribosomes, and numerous dictyosomes. The role of the dictyosomes is twofold: 1. dictyosome vesicles bud coated vesicles which transport materials from the cell and, 2. dictyosome vesicles coalesce, forming large storage vesicles. The storage vesicles are surrounded by, and often in contact with, poculiform RER. The RER forms an interconnected network throughout the cytoplasm, extending from the nuclear envelope to the plasmalemma. Distended profiles of RER are frequently in direct contact with the plasmalemma. Thus, inPharbitis secretory trichomes, it is the coated vesicles and RER which are active in secretion export. These findings imply a secretory pathway which deviates from the usual pattern in glandular cells.Predoctoral fellow of National Science Foundation during part of the investigation.     

Fine structure of the adenohypophysis in immature sockeye salmon,Oncorhynchus nerka

Summary The ultrastructure of the secretory cells of the adenohypophysis of juvenile sockeye salmon was investigated. Pituitary glands were collected from immature fish transferred experimentally to sea water and subsequently returned to fresh water. The rostral pars distalis contained three cell types: ACTH cells, prolactin cells, and non-secretory cells. The prolactin and non-secretory cells were joined together in the form of follicles by desmosomes and they both had cilia and microvilli projecting into the follicle lumen. Various follicular structures such as lumen, multivesicular structures, and peripheral basement membrane are discussed as possible sites of prolactin cell granule release. The columnar ACTH cells were found at the junction of the rostral pars distalis and the neurohypophysis. The cytoplasmic granules in these cells were characteristically separated from their limiting membrane by a clear space. Multivesicular structures were also found in association with this cell type. The caudal pars distalis also contained three cell types: one acidophil (putative somatotrop) and two basophils (putative thyrotrops and gonadotrops), all of which were similar to those described in adult fish. The pars intermedia contained only one cell type. They appeared to be active cells and were characterized by containing membrane-bounded granules similar to those found in the ACTH cells. Changes in ambient salinity had no apparent effect on any cell type described.The work was supported by a grant in aid of research from the National Research Council of Canada. We wish to thank Mr. R. Lindsay, Mr. C. Cooper, and Mr. G. Longworth for their technical assistance. We would also like to thank Mr. S. Killick of the International Pacific Salmon Fisheries Commission for his assistance in the collection of fish and Dr. H. Cook for his helpful discussion of the project. This paper is No. 058 in the University of Guelph Migration Series.     

The ultrastructural basis for the identification of cell types in the pancreatic islets I. Guinea pig

Summary The pancreatic islets of the guinea pig have been studied by light and electron microscopy. The B granules in glutaraldehyde-fixed tissue often are cup-shaped with an indentation visible on one side of the granule. Phosphotungstic acid hematoxylin (PTAH) positive cells have been characterized by electron microscopy as three subtypes based on the size of the secretory granules. A_a cells are the most common and have secretory granules around 200 m in diameter. A_b cells have large secretory granules around 300 m in diameter and are relatively infrequent. A_c cells are the least common and have small (160 m) granules. Characteristic D cells are identifiable by electron microscopy and, on the basis of the subsequent study (Munger, Caramia, and Lacy, 1965), are identified as the silver positive cells observed by light microscopy.This investigation was supported in part by United States Public Health Service research grants GM-10102 and GM-03784 from the Institute of General Medical Sciences, and AM-01226 from the Institute of Arthritis and Metabolic Diseases.-The authors wish to acknowledge the valuable technical assistance of Mrs. Aileen Sevier and Mrs. Lidia Donohue.     

Ultrastructure of the human submaxillary gland

Summary Myoepithelial cells in the human submaxillary gland are stellate in form, with long, tapering processes. They are interposed between the base of the secretory cells and the basement membrane, and are bound to the secretory cells by desmosomes. Their cytoplasm contains numerous myofilaments measuring approximately 40 Å in diameter, which frequently aggregate to form structures similar to the dark bodies seen in smooth muscle cells. The myofilaments are anchored to the plasma membrane by attachment devices. The myoepithelial cells are often accompanied by cells which have a similar shape, but possess an extremely electron-lucent cytoplasm that contains almost no organelles or inclusions. Electron microscopical observations indicate that these cells, termed clear cells, are transformed directly into myoepithelium, since all morphological intergrades between the two cell types have been recognized.This work was supported in part by a grant from the Henry Spenadel Trust. The technical assistance of Mr. Roy R. Keppie and Mrs. Mona Brandreth is gratefully acknowledged.     

Control of the pars intermedia of the lizard,Anolis carolinensis

Summary The ultrastructure of the intermediate lobe of the hypophysis was studied in Anolis carolinensis with the use of a threefold aldehyde fixative. Lizards with a brown skin were selected. The possibility of two types of secretory cells is discussed; neither cell type is innervated. Type I cells are rarely found and contain dense granules approximately 0.3 m in diameter; Type II cells vary widely in secretory activity. Most of the Type II cells contain a large number of dense secretory granules (up to about 1.3 m in diameter) almost filling the cytoplasm. Rough endoplasmic reticulum (RER), Golgi apparatus and mitochondria are poorly developed. Only some of these cells show signs suggesting a high secretory activity, namely a well developed RER, Golgi apparatus and numerous mitochondria. In these cells the RER sometimes forms large intracisternal droplets (up to 7 m in diameter). Two of the animals exhibited a more uniform, high secretory activity. Large (about 2 m in diameter), pale vacuoles, probably of extracellular character, were found mostly in the vicinity of the perivascular septum. Their role in the release of MSH is discussed. The present data, which are discussed with reference to earlier findings (Forbes, 1972), form the morphological basis for an experimental study on regulation of MSH release (Larsson et al., 1979).Supported by grants from the Swedish Natural Science Research Council (to P. Meurling) and the Royal Physiographic Society of LundThe authors are indebted to Mrs. Ingrid Hallberg, Mrs. Kirsten Thörneby and Mrs. Lena Sandell for valuable technical assistance and to Miss Inger Norling for photographic aid     

The pituitary of Aphanius dispar (Rüppell) from hypersaline marshes and freshwater

Summary An ultrastructural study of the rostral pars distalis of the pituitary of Aphanius dispar specimens taken from freshwater or hypersaline marshes revealed significant structural differences which indicate higher activity of the prolactin cells in the hypotonic medium. Prolactin cells from freshwater specimens had larger secretory granules, a higher amount of endoplasmic reticulum, and expanded intercellular spaces with many secretory lakes. These cells contained an unusual cytoplasmic structure, consisting of twisted canals with vesicular lumina, connected to the endoplasmic reticulum of the cell. This structure is about 1–2 m in diameter.Stellate cells are characterized by extracellular spacing junctions which are particularly noticeable at the confluence of the interstellate cell canaliculi and the pericapillary space.Abbreviations FW freshwater - HS hypersaline - NS neurosecretory - PCB paracrystalline body - PNH proximal neurohypophysis - RPD rostral pars distalis - SG secretory granule - SW seawater This paper is dedicated with affectionate respect to Professor Berta Scharrer on the occasion of her 70th birthdayThe assistance of Cynthia Bellon in editing this paper is gratefully acknowledged     

Ultrastructural studies on prolactin and growth hormone cells in Anguilla pituitaries in vitro

Summary Eel hemi-pituitaries were cultured in vitro on high or low sodium media which are known to affect differentially prolactin and growth hormone release. Ultrastructural examination of the prolactin cells after 24 h culture showed the Golgi bodies were markedly more abundant and widely distributed in hemi-pituitaries from the low sodium medium. Secretory granule release profiles and dense bodies were also more frequent, but the percentage of the cytoplasmic volume occupied by secretory granules was lower than on the high sodium medium. RER was only slightly modified. Significant differences were noted in the shape and processes of the non-granulated (stellate) cells of the RPD, but there were only slight differences in the ultrastructure of the somatotropes.     

Fine structure of the gastric epithelium of the ascidian botryllus schlosseri vacuolated and zymogenic cells

Summary Vacuolated and zymogenic cells, which are two of five cell types identified by electron microscopy in gastric epithelium of B. schlosseri, are described in detail. The vacuolated cells are characterized by one, or a few, supranuclear vacuoles containing myelin figures. A peculiar Golgi apparatus is consistently found at the base of the vacuoles; it consists of cisternae frequently containing small vesicles and tubules of constant diameter and/or a strong electron-opaque material. A variety of vesicles and multivesicular bodies are visible in the apical cytoplasm below long ribbon-like microvilli. The se findings suggest that the vacuolated cells are involved in absorptive and perhaps secretory activity. The zymogenic cells are characterized by a highly developed RER, numerous apical secretory granules and a well developed supranuclear Golgi apparatus. At the apical end, autophagosomes are frequently encountered, some of which contain also zymogen granules. Both cell types contain numerous lipid droplets, which are interpreted as an energy reserve available for the cells and for the entire colony during the change of generation. Correlation between structure and function in both cell types is discussed by taking into account the peculiar life cycle of B. schlosseri, as well as previously reported data on similar cells in other ascidians.We would like to dedicate this work to Prof. Giuseppe Reverberi on the occasion of his 70th birthday.The authors are indebted to Profs. A. Sabbadin and G. Mazzocchi for their most helpful suggestions and advice. We would also like to thank Mr. G. Tognon for technical assistance and the staff of the Stazione Idrobiologica di Chioggia for their assistance in collecting material. — This research was supported by a grant of C.N.R., contract from the Istituto di Biologia del Mare, Venezia, No. 7100396/04115542 and with the E.M. facilities of C. N. R. contract No. 70.01798.04.115.876.     

Electron microscope studies of the intracellular origin and formation of calcifying granules and calcium spherites in the hepatopancreas of the snail,Helix pomatia L.

Summary It has been previously demonstrated that the hepatopancreas of the snail, Helix pomatia, produce proteinaceous particles (termed b-granules), which are involved in the process of calcification. In calcium cells of this organ calcium spherites arise from these granules. The present study retraces the intracellular development of b-granules from Golgi vesicles and from Golgi saccules encircling the cytoplasmic areas up to the formation of calcium spherites. It is suggested that the mature b-granules are well-defined cytoplasmic units with calcifying capacity. The fine structure of b-granules and calcium spherites is described. The possible function of various cell organelles in the formation of the b-granules and spherites is discussed.This investigation was supported by grants from the Swedish Natural Science Research Council. The assistance of Mrs. I. Rehnberg is gratefully acknowledged.     

Further freeze-fracture studies on the uterine epithelium of Salamandra salamandra (L.) (Amphibia,Urodela) using the antibiotic filipin

Summary The apical portion of the uterine lining of the ovoviviparous fire salamander, Salamandra salamandra, was studied by the freeze-fracture technique in conjunction with the polyene antibiotic filipin. Filipin-sterol complexes were found in the luminal plasmalemma and in the membranes limiting the mucous secretory granules typical of this epithelium. In all females, but particularly in non-pregnant females, more or less discrete clusters of filipinsterol complexes were occasionally found overlying heavily affected secretory granules. The findings are discussed with regard to comparable results (Orci et al. 1980) based on the examination of collapsed and stretched urinary bladders of toads.We are indebted to Mrs. K. Ott for excellent technical assistance, to Miss E.S. MacLure for linguistic corrections and to Dr. J.E. Grady of the Upjohn Co., Kalamazoo, Michigan USA, for kindly providing the filipin     

The fine structure of barley aleurone cells

Summary The ultrastructural morphology of both dry and water-imbibed barley aleurone cells is described. The aleurone cell is characterized by the presence of numerous aleurone grains and spherosomes. In addition, it contains organelles typical of other plant cells including structures similar to microbodies, and rough endoplasmic reticulum characterized by the presence of numerous polyribosomes. It is inferred that the morphological specialization of aleurone cells is related to their biochemical specialization.Work supported by National Science Foundation grant GB5863. The skillful technical assistance of Mrs. Janet Price is gratefully acknowledged.     

Lysosomes in the cessation of vitellogenin secretion by the mosquito fat body; selective degradation of Golgi complexes and secretory granules

A massive and selective degradation of Golgi complexes, secretory granules, and RER is the mechanism responsible for the rapid termination of Vg secretion by trophocytes of the mosquito fat body. These cells are involved in an intensive synthesis of a glycoprotein, vitellogenin (Vg), which is accumulated by developing oocytes as yolk protein. Previously, assays for lysosomal enzymes have demonstrated that the cessation of Vg synthesis is characterized by a sharp increase in lysosomal activity; and fluorescent microscopy has shown that, during this intense lysosomal activity, Vg concentrates in lysosomes. In this report, electron microscopy combined with cytochemistry for lysosomal enzymes and localization of Vg with colloidal gold immunocytochemistry has shown that this lysosomal activity is directed towards selective degradation of Vg and organelles associated with its synthesis and secretion. Three organelles undergo lysosomal breakdown: the Golgi complex, Vg-containing secretory granules, and RER. The degradation of Golgi complexes occurs in two steps similar to that for RER: first, the organelle is sequestered by double isolation membranes, and the resulting pre-lysosome then fuses with a primary or secondary lysosome. In contrast, mature Vg-containing secretory granules fuse with lysosomes directly. This combination of crino- and autophagy is a specific, highly intense, and precisely timed event.     

Structure of the nongranulated cells in the hypophyseal rostral pars distalis of cyclostomes and actinopterygians

Summary The structure of the nongranulated cells in the sea lamprey adenohypophysis and similar cells of the rostral pars distalis in a number of actinopterygian fishes was examined with the aim of determining the role(s) of these cells in pituitary function.A number of possible roles are proposed for the nongranulated cells. In salmonids and Amia calva the cells may be involved in the active movement of material into or from the follicle lumina. The structure of the nongranulated cells in in vitro cultured and in in vivo transplanted trout rostral pars distalis also suggests a phagocytotic role for these cells. In teleosts with a non-follicular rostral pars distalis the nongranulated cells appear to play roles in the release of granules from the prolactin cells and in the subsequent dispersal of the hormone (and/or carrier substance) into the peripheral circulation.We wish to thank Dr. B.I. Baker who performed the trout pituitary in vitro cultures and Professor M. Hyder who was closely involved with the collection of the Tilapia specimens. We also wish to thank Mr. R. Lindsay and Mrs. L. Lin for their technical assistance and Drs. F.W.H. Beamish and R. Sonstegard for their help in supplying some of the animals used in this study. —The work was supported in part by a grant-in-aid of research from the National Research Council of Canada to J.F.L. and in part by a negotiated NRC grant. The paper is number 138 in the physiology of migration series.     

The ocellus of the cockroach,Periplaneta americana (Blattariae)

Summary The ocelli of Periplaneta americana were studied by light and electron microscopy. The view that the ocellus of the cockroach represents a degenerated structure can no longer be supported. All organelles necessary for function are present.The club-shaped retinular cells lie homogeneously distributed in the cupule of the ocellus. Rhabdoms are seen as sickle-, y-, x-or star-shaped structures with up to six cells in formation. Cells were found which had formed two rhabdomeres.The mass of cell organelles lies in the cytoplasm between the cell nucleus and the rhabdom. Smooth endoplasmic reticulum is wound into a spindle formation of considerable size at the origin of the axon in some cells. A cylindrical body in which 10–40 microtubules are packed, as yet unknown in insect retinular cells, is described.The receptory area of the ocellus terminates in a tapetum which contains granules, soluble in alcohol. The axon bundles of the retinular cells run through the tapetum and immediately thereafter make synaptic contact with dendrites of the ocellar nerve cells, while still within the ocellus.The authors are indebted to Mrs. Margaret Weber-Wood for her linguistic assistance