Electron microscopic studies on the adenohypophysis of the White-Crowned sparrow,Zonotrichia leucophrys gambelii

Summary The pars distalis of the adenohypophysis of normal (78), thyroideotomized (6), adrenalectomized (6), and castrated (14) White-crowned Sparrows were observed with the electron microscope. Six types of glandular cells were identified and the ultrastructural characteristics of each have been described. To each has been assigned tentatively an endocrine function.STH cells are characterized by the presence of large, dense secretory granules ranging from 220–280 m, a poorly developed endoplasmic reticulum, and a fragmented Golgi apparatus; they occur only in the caudal lobe. They show no remarkable changes after adrenalectomy, castration, and thyroidectomy.Prolactin cells, whose identity is suggested by their responses to photostimulation and surgical experiments, are characterized by large, polymorphic, dense secretory granules; they have been found mainly in the cephalic lobe.ACTH cells, whose function is confirmed by their cytological responses to adrenalectomy, have a peculiar type of secretory granule (220 m) with high and low phases of electron density. They occur exclusively in the cephalic lobe and are transformed, after adrenalectomy to large, vacuolated adrenalectomy cells.TSH cells are so designated by their response to thyroidectomy. After thyroidectomy, they lose their specific fine secretory granules and are transformed into large, vacuolated thyroidectomy cells. We have found TSH cells and thyroidectomy cells only in the cephalic lobe.Two types considered to be gonadotropic cells from their responses to gonadectomy, occur in both the cephalic and caudal lobes. One of them contains spherical, dense secretory granules (180–220 m), prominent rough endoplasmic reticulum and a well developed Golgi apparatus; the other type contains dense secretory granules of variable size (150–350 m), a less extensively developed Golgi apparatus, and sac-like endoplasmic reticulum. Both types of gonadotropic cells show extreme enlargement and vacuolization after castration. However, they retain differences in appearance in the structure of cytoplasmic organelles and vacuolization.Dedicated to Professor Dr. H. Grau in honor of his 70th birthday.The investigation reported herein was supported by a research grant (HE 07240 NEUA) from the National Institutes of Health to Professor Vitums, by a research grant (5R01 NB 06187) from the National Institutes of Health to Professor Farner, and by a scientific research grant (No. 91049) from the Ministry of Education of Japan to Professor Mikami. The authors wish to thank Professor James R. King for his assistance in obtaining and maintaining the birds, and for his helpful advice concerning the experiments.     

Cytodifferentiation of the adenohypophysis of the domestic fowl

Summary In the chick embryo the first membrane-bound secretory granules occur in the cytoplasm of occasional cells in the cephalic lobe of pars distalis at the 7th day of incubation. On the 8th day most of the cells in both the cephalic and caudal lobes contain secretory granules that are variable in size, form and density.On the 9th day at least two types of glandular cells are distinguishable in the cephalic and in the caudal lobes; however, these cells are not comparable with those of the adult gland. Differentiation of acidophils and basophils occurs, apparently simultaneously, in 11-day embryos.The cells of the cephalic and caudal lobes are morphologically distinct from their first appearance. Thus it is concluded that these two lobes develop independently and differently from an early stage of ontogenesis.The secretory granules are formed in the Golgi area of the hypophysial cells after the 8th day of incubation. However, secretory material may be synthesized also by a process not involving the Golgi apparatus.Nerve fibers containing granules first appear in the superficial layer of the median eminence on the 8th embryonic day and by the 12th day three types of granules and two types of clear vesicles are identifiable.The investigation reported herein was supported by grant from Japan-U.S. Cooperative Science Program of Japan Association for Science Promotion to Professor Mikami and by U.S.-Japan Cooperative Science Program Grant No. GF-33334 to Professor Farner.     

The relationship between the folliculo-stellate network and the thyrotropic cells of the avian adenohypophysis

Summary The folliculo-stellate network of the avian adenohypophysis consists of stellate cells surrounding colloid-containing follicular cavities into which cilia and microvilli project. Other identifying criteria are agranularity, junctional complexes at the apical pole, presence of cytoplasmic processes ramifying between adjacent secretory cells, and close appositions of plasma membranes linking folliculo-stellate cells and presumptive thyrotropic cells.Transmission electron microscopy reveals that TRH and L-DOPA induce simultaneous ultrastructural changes in the folliculo-stellate network and in the thyrotropic cells. TRH transforms at cell of the cephalic lobe into a highly hypertrophic cell in which enlargement of cisterns of rough endoplasmic reticulum containing secretory granules, development of a large Golgi complex, presence of newly synthesized secretory granules, and granulation of the cytoplasm are the main features. In the meantime, the follicular cavities become dilated by large amounts of homogeneous colloid. The administration of L-DOPA also leads to the development of dilated cisterns in presumptive thyrotropic cells of the cephalic lobe. Intracisternal granules, immature secretory granules, and large Golgi complexes, however, are not observed. Degranulation of the cytoplasm is obvious. The follicular cavities of both cephalic and caudal lobes are enlarged and filled with colloid in which granular elements are noted.The ultrastructural changes observed in thyrotropic cells and in the folliculo-stellate network reflect functional changes induced by the experimental manipulation. These changes may be related, directly or indirectly, or completely independent.     

Cell types in the adenohypophysis of the Japanese quail and effects of injection of luteinizing hormone-releasing hormone

The cells of the adenohypophysis of the Japanese quail were studied by both light and electron microscopy after exposure to long photoperiods or injection of lutenizing hormone-releasing hormone (LRH). Six cell types were identified in the adenohypophysis by examining alternate thick and thin sections by light and electron microscopy. In the cephalic lobe, there are four types of glandular cells. They are the prolactin cells, ACTH cells, TSH cells, and gonadotropic cells (FSH?). In the caudal lobe, there are two types of cells, STH cells and gonadotropic cells (LH?). After exposure to long daily photoperiods, gonadotropic cells in both lobes were strongly activated. They became larger and accumulated many granules. ACTH cells became vacuolated; granules were sparse. Synthetic LRH injection (10 mug/0.2 ml/day) for 10 days to the non-photostimulated quail stimulated certain numbers of the gonadotropic cells in the both lobes, although the response of the cells was less than that induced by photostimulation. No response was seen in the other cell types.     

Immunofluorescent localization and ultrastructural characterization of gonadotrophe cells in the adenohypophysis of the barbary drake (Cairina moschata L.) using anti-chicken LH serum

Summary An indirect immunofluorescence technique and an anti-chicken LH serum were used to localize cells in the adenohypophyses of drakes at different stages of their breeding cycle, after castration, and after castration combined with thyroxine treatment. Immunofluorescent cells were distributed throughout both lobes of the adenohypophyses from control and experimental birds and were shown to be alcian blue positive, PAS negative, basophiles. Immunofluorescent cells were as numerous in castrated birds as in castrated birds treated with thyroxine. Adjacent thin and semi-thin sections were used to study the cells binding anti-LH serum at light microscope and ultrastructural levels. The cells contained spherical granules with variable densities and diameters ranging between 40 and 280 nm in the rostral (= cephalic) lobe, and between 60 and 260 nm in the caudal lobe. The light microscope and ultrastructural observations showed that the anti-LH serum binds to cells which have been classified by other authors in the Pekin duck, quail and pigeon as TSH producing delta cells. The experimental technique used did not permit a distinction to be made between cells producing FSH and LH.Supported by DGRST (Contrat n 75-7-0803 A.C. Biologie de la Reproduction et du Développement)     

Light and electron microscopic investigations of six types of glandular cells of the bovine adenohypophysis

Summary Twelve bovine adenohypophyses were prepared for light and electron microscopy of the cell types of pars distalis. Correlation between the light and electron microscopy was effected by use of alternate thin and thick sections. Cytological changes in the experimental animals were used as criteria for the identification of six different types of secretory cells.Two types of acidophils, alpha and epsilon cells, are recognized in peripheral area of the pars distalis by light and electron microscopy. The alpha cells contain orangeophilic secretory granules of a maximum diameter of 400–450 m and correspond to ordinary acidophils (STH cells). The second type, epsilon cells, contains larger, fuchsinophilic granules of 600 to 900 m in diameter, increase in number and granulation after pregnancy and thyroidectomy, and are thought to be prolactin cells (LTH cells).Two types of amphophils, zeta and delta 1 cells, were found in the central area of the pars distalis. The zeta cells contain smaller numbers of amphophilic, cored granules (200 m maximum diameter) and based on the comparison with literature on other species of animals, are designated as ACTH cells. The delta 1 cells are round or oval and contain very dense, spherical granules (250–300 m) which are stained red or reddish purple with PAS, aldehyde thionin and PAS-methyl blue methods. They show extreme enlargement and bizarre cytoplasmic appearance after castration and are designated tentatively as LH gonadotrophs or LH cells.Two types of basophils, beta and delta 2 cells, were also identified by correlative light and electron microscopy. The beta cells are polygonal in outline, distributed exclusively in the zona tuberalis and contain large, less dense secretory granules (300–400 m) which are stained selectively with Gomori's aldehyde fuchsin. After thyroidectomy, they lose their secretory granules and are transformed into large, vacuolated thyroidectomy cells. They are therefore, identified as thyrotrophs or TSH cells. The delta 2 cells are round, oval or polygonal in shape and contain basophilic granules ranging from 220 to 300 m in diameter. They show extreme enlargement and vacuolization due to the dilation of endoplasmic reticulum, after castration, and are designated tentatively as FSH gonadotrophs or FSH cells.The investigation reported herein was supported by a Scientific Research Grant (No. 291049) from the Ministry of Education of Japan.     

Ultrastructural study of the adenohypophysis of the Chinese hamster.

The adenohypophysis of normal Chinese hamsters of both sexes was examined ultrastructurally. Organs were fixed by intravascular perfusion with S-collidine-buffered glutaraldehyde solution. Seven types of cells were differentiated and, according to morphological characteristics, classified as (1) mammotropes, with very large (400-800 nm) polymorphous secretory granules; (2) somatotropes, either in the storage phase with numerous large, dense granules (average 300 nm), or in the hormone synthesis phase, with abundant endoplasmic reticulum and large Golgi apparatus; (3) corticotropes, with irregular cell shape, and granules (average 160 nm) arranged in lines parallel to the cell membrane; (4) FSH gonadotropes, with abundant and dilated endoplasmic reticulum, and granules (190-320 nm) uniformly distributed in the cytoplasm; (5) LH gonadotropes, with granules (120-220 nm) of varied density; (6) thyrotropes, with irregular cell shape and very small granules (120-160 nm), and (7) agranulated cells. The ultrastructure of the adenohypophysis of the Chinese hamster corresponds closely with observations reported in rats, mice and Syrian hamsters.     

Investigations of the butylcholinesterase-containing cells of the adenohypophysis of the white-crowned sparrow,Zonotrichia leucophrys gambelii

Summary In the adenohypophysis of Zonotrichia leucophrys gambelii two types of cells with butylcholinesterase(BuChE) activity can be demonstrated histochemically. Type I occurs in the cephalic lobe of the pars distalis and in the pars tuberalis; it consists of oval and round cells. It is a distinctive cell type that is identical with the amphophilic cells described by Matsuo, Vitums, King and Farner (1969). Whereas castration or inhibition of thyroid gland activity causes only minor changes in these cells, blocking of adrenal cortex activity, or adrenalectomy, causes conspicuous hyperplasia and hypertrophy suggesting that these cells are involved in the production or release of ACTH. The second type, which occurs in both cephalic and caudal lobes, consists mostly of irregularly formed cells. Various observations indicate that it is a composite group, consisting, at least in part, of degenerating cells.These investigations were supported by grants from the National Institutes of Health (S ROI NB 06812) and the National Science Foundation (GB 5969) to Professor Farner.     

The prolactin cell of the white-crowned sparrow,Zonotrichia leucophrys pugetensis

Summary The anterior pituitaries from a series of female White-crowned Sparrows,Zonotrichia leucophrys pugetensis, in the periods of oviposition, incubation, and brooding under natural conditions, have been investigated by electron microscopy. The prolactin cells occur in cephalic lobe and are characterized by large (ca. 300–600 m), polymorphic electron-dense secretory granules and an extremely well developed, lamellated endoplasmic reticulum. During incubation and brooding it is only these prolactin cells that are in an activated secretory phase, as indicated by increase in number and size, extremely well developed endoplasmic reticulum, decrease in number of mature secretory granules, and by active formation of granules in the enlarged Golgi apparatus. In the late stages of brooding, and post-breeding, the prolactin cells regress with involution of the endoplasmic reticulum and Golgi apparatus, reaccumulation of granules, and the appearance of lysosomes.The gonadotropes of both the cephalic and caudal lobes undergo progressive morphologic changes through the course of the breeding period. They are numerous and active in the ovulating bird. They undergo gradual regression during the periods of incubation and brooding to become typical broody cells.This investigation was supported by Grant No. GF-33334, U.S.-Japan Cooperative Science Program of the National Science Foundation and by Grant No. GB-28080X, also from the National Science Foundation, to Professor Farner; and by Grant No. 5R040 Japan-U.S. Cooperative Science Program of Japan Association of Science Promotion, to Professor Mikami.     

Ultrastructure of the acinar cells in the submandibular gland of the nine-banded armadillo

There are two discrete lobes comprising the armadillo subman-dibular gland. These two lobes can be defined grossly, histochemically and morphologically with the light and electron microscope. The minor lobe stains more intensely with PAS and AB. When viewed in the electron microscope, the secretory granules of the acinar cells within this lobe appear mucous-like. The granules of the demilune cells are slightly different in appearance. The secretory granules of the acinar cells in the major lobe contain many dense foci embedded in a fibrillar matrix, a substructure not described previously. The demilune cells of this lobe contain secretory granules with a mucous-like structure which is consistent throughout the entire lobe. As in the minor lobe, these demilune cells stain very intensely with PAS and AB.     

Morphological changes of pituitary gonadotrophs and thyrotrophs following treatment with LH-RH or TRH in vitro

Summary To investigate the relationship between LH-RH and TRH and the formation of castration cells and thyroidectomy cells, pituitary glands of 14-day old female rats were cultured with LH-RH or TRH for 10 days. Observed in these glands were external and internal zones, the former containing active, healthy appearing cells, and the latter consisting of many degenerative and necrotic cells. Gonadotrophs and thyrotrophs were readily demonstrated in the external zone of the organs by immunocytochemistry. Hypertrophie gonadotrophs (castration cells) and thyrotrophs (thyroidectomy cells) were observed only in the external zone by electron microscopy. Neither the typical signet-ring gonadotrophs nor intracisternal granules of thyroidectomy cells were seen in the present study. However, the hypertrophic gonadotrophs or thyrotrophs were similar to castration cells or thyroidectomy cells observed in the pituitary gland following castration or thyroidectomy. The results indicate that LH-RH and TRH directly stimulate the secretion of hormones and alter the morphological features of pituitary target cells.Supported by USPHS Grant HD 11826. The author wishes to express his thanks to NIAMDD for providing pituitary hormones     

Development of gonadotropes in the chicken embryonic pituitary gland

Although a number of immunohistochemical studies have been carried out on the differentiation of chicken gonadotropes during embryogenesis, the temporal and spatial properties of appearance of gonadotropes are not clear. In this study, we studied the appearance and morphological characteristics of gonadotropes in the embryonic and adult chicken anterior pituitary glands using RT-PCR, in situ hybridization and immunohistochemistry. For this purpose, we raised specific antisera against chicken follicle-stimulating hormone beta-subunit (cFSHbeta) and chicken luteinizing hormone beta-subunit (cLHbeta) based on each putative amino acid sequence. RT-PCR analysis revealed that cFSHbeta mRNA was expressed from embryonic day 7 (E7). Chicken FSHbeta mRNA-expressing (-ex) and -immunopositive (-ip) cells started to appear in the ventral part of the caudal lobe in the anterior pituitary gland at E8. Chicken LHbeta-ip cells were also first observed there at E8, but cLH mRNA expression was confirmed from E4 by RT-PCR analysis. The distribution of these chicken gonadotropin-ex and -ip cells spread from the ventral part to dorsal part in the caudal lobe around E10 and subsequently expanded to the cephalic lobe from E12 to E20. These cells were morphologically classified into two types (round- and club-shaped cells). It was found that the density of gonadotropin-ip cells in the caudal lobe was always higher than that in the cephalic lobe throughout the period of development. To the best of our knowledge, this is the first report focusing on the differentiation of chicken gonadotropes by assessment of both protein and mRNA of chicken gonadotropin.     

Immunohistochemistry of the hypothalamic neuropeptides and anterior pituitary cells in the Japanese quail

The pars distalis of the avian adenohypophysis consists of well-defined cephalic and caudal lobes which are distinct in their cellular constituents. Immunocytochemical investigations on the pituitary hormones of the pars distalis of the Japanese quail reveal five types of secretory cells, adenocorticotropin (ACTH) cells, prolactin (PRL) cells, thyroid-stimulating hormone (TSH) cells, growth hormone GH (STH) cells, and FSH/LH (gonadotropic) cells. The ACTH cells, TSH cells, and PRL cells are restricted to the cephalic lobe, and GH (STH) cells are confined to the caudal lobe, while FSH/LH cells are distributed throughout the cephalic and caudal lobes. The median eminence of birds has distinct anterior and posterior divisions, each with different neuronal components. The avian hypophysial portal vessels also consists of two groups, anterior and posterior. The peculiar arrangement and distribution of the avian hypophysial portal vessels are possibly related to the distribution of neuropeptides in the two divisions of the median eminence and to the cytological and functional differentiation of two lobes of the pars distalis. The localization of perikarya and fibers containing luteinizing hormone releasing hormone (LHRH), somatostatin, vasotocin, mesotocin, corticotropin-releasing factor (CRF), vasoactive intestinal polypeptide (VIP), glucagon, metenkephalin, and substance P in the hypothalamus and median eminence of the Japanese quail has been investigated by means of immunohistochemistry using antisera against the respective neuropeptides. LHRH-, somatostatin-, VIP-, met-enkephalin-, and substance P-immunoreactive fibers are localized in the external layer of the anterior and posterior divisions of the median eminence, while CRF- and vasotocin-reactive fibers are demonstrated only in the external layer of the anterior division of the median eminence. The metenkephalin fibers are thicker in the anterior median eminence but the substance P fibers are more abundant in the posterior division. Mesotocin fibers occur only in the internal layer of the median eminence and neural lobe.     

Morphological and biochemical analyses of the salivary glands of the malaria vector, Anopheles darlingi.

Adult Anopheles darlingi salivary glands are paired organs located on either side of the esophagus. The male glands consist of a single small lobe. The female gland is composed of two lateral lobes, with distinct proximal and distal portions, and a medial lobe. The lobes are acinar structures, organized as a unicellular epithelium that surrounds a salivary canal. The general cellular architecture is similar among the lobes, with secretory material appearing as large masses that push the cellular structures to the periphery of the organ. Cells of the proximal-lateral lobes show asynchronous cycles of secretory activity and contain secretory masses with finely filamentous aspect. In the distal-lateral lobes, cells display synchronous cycles of activity, and have a dense secretory product with mottled pattern. Cells of the medial lobe have secretory masses uniformly stained and highly electrondense. Biochemical analysis of the adult female salivary glands revealed apyrase, alpha-glucosidase and lysozyme activities. Alpha-glucosidase and lysozyme activities are detected mostly in the proximal lobes while apyrase is mainly accumulated in the distal lobes. This differential distribution of the analyzed enzymes reflects a specialization of different regions for sugar and blood feeding. Thus, the morphological differences observed in the lobes correlate with functional ones.     

Immunohistochemical and ultrastructural study of anterior pituitary cells in the female Afghan pika,Ochotona rufescens rufescens

An immunohistochemical study of the anterior pituitary gland of the female Afghan pika was carried out to distinguish the ultrastructural features of GH, PRL, ACTH, TSH and LH cells. The histochemically identified GH cells resembled ultrastructurally oval or round GH cells of the rat laden with large, dense secretory granules. PRL cells were divided into three subtypes based on differences in the diameter of their spherical secretory granules. They lacked polymorphic or irregularly shaped secretory granules. ACTH cells resembled ultrastructurally, in some respects, Siperstein's "corticotrophs" of the rat with peripheral arrangement of secretory granules. However, they were not always stellate, but elongate or angular in shape. The dense secretory granules were concentrated in the peripheral area of cytoplasm. TSH cells were non-stellate, but usually oval in shape, containing the smallest spherical secretory granules (100-200 nm in diameter). Almost all LH cells reacted also with FSH antiserum. They were irregular in shape, sometimes in contact with or surrounded the GH cells. They contained an abundance of medium-sized secretory granules (140-260 nm in diameter) which were larger than those in the LH cells of the female rat throughout the estrous cycle. Large secretory granules in the LH cells of the female pika seemed to be related to the endocrine state of persistent estrus.     

Independent differentiation of mammotropes and somatotropes in the chicken embryonic pituitary gland

It has been reported that mammotropes in a rodent pituitary gland are derived from somatotropes via somatomammotropes (SMTs), cells that produce both growth hormone (GH) and prolactin (Prl). However, no studies have been done on the transdifferentiation of somatotropes in the chicken pituitary gland. In this study, in order to determine the origin of mammotropes, we studied detail property of appearance of chicken somatotropes, mammotropes and pit-1 cells and then evaluated the existence of SMTs in the chicken embryonic pituitary gland. Immunohistochemical analysis revealed that GH-immunopositive (GH-ip) cells appeared on embryonic day (E) 14 and were mainly distributed in the caudal lobe, while Prl-immunopositive (Prl-ip) cells appeared in the cephalic lobe of the pituitary gland on E16. In situ hybridization (ISH) and RT-PCR analysis showed that expression of GH and Prl mRNA starts at E12 in the caudal lobe and at E14 in the cephalic lobe respectively. Pit-1 mRNA was first detected on E5 by RT-PCR, and pit-1 mRNA-expressing cells were found in the cephalic lobe on E8. Then with the ontogeny of the chicken, these cells spread into both lobes. Using a double staining method with ISH and immunohistochemistry, we could not detect the existence of SMTs in the chicken embryonic pituitary gland even in the marginal region of either lobe. These results suggest that chicken somatotropes and mammotropes independently appear in different lobes of pituitary gland and that transdifferentiation from somatotropes to mammotropes is not the central route for differentiation of mammotropes in the embryonic chicken pituitary gland.     

Light-microscope studies of the cytology of the adenohypophysis of the white-crowned sparrow,Zonotrichia leucophrys gambelii

Summary Adenohypophyses from more than two hundred white-crowned sparrows of both sexes and different ages, and from different periods of their annual reproductive cycle, have been used for this investigation. In addition to examination of these normal birds, we have also studied the adenohypophyses of 23 castrates and 24 controls held in different photoperiodic conditions.Cytologically the pars distalis of the adenohypophysis of the white-crowned sparrow is typically avian with distinct cephalic and caudal lobes, each with characteristic cell-types.Four basic cell-types, the acidophils, basophils, amphophils, and chromophobes, have been identified in the pars distalis by means of Matsuo tetrachrome and Matsuo modified PAS-methyl blue staining methods.Three types of acidophils, orange, red, and small, are confined to the caudal lobe of the pars distalis. Their possible functions are discussed.Light basophils (PAS-light red cells) and deep basophils (PAS-deep red cells) are equally distributed in both lobes. It is suggested that basophils may be involved in gonadotropic function since their appearance correlates well with the annual gonadal cycle and photoperiodic stimulation of gonadal growth and with the results of castration.The amphophils or PAS-purple cells (aldehyde-fuchsin positive) are found only in the cephalic lobe. Their probable function is discussed.Two types of chromophobes, specific and ordinary chromophobes, have been observed. The specific chromophobes are found only in the cephalic lobe and are similar to the Kernhaufen described by Romeis (1940). The ordinary chromophobes are similar to those of the pars distalis of other avian species and of mammals.The castration cells are found in both lobes of the photosensitive castrates under natural photoperiodic conditions as well as in those subjected artificially to photostimulation (20-hour daily photoperiods). Similar cells have also been observed in the pars tuberalis of the castrated photostimulated birds.The relations of the rostral and caudal groups of the portal vessels to the cell-types found in the cephalic and caudal lobes are discussed.Dedicated to Professor Dr. Y. Kato, Department of Anatomy of the Domestic Animals, Kyushu University, Fukuoka, Japan, and to President Dr. H. Mimura, Shinshu University, Matsumoto, Japan, in honor of their retirement.The investigation reported herein was supported by a research grant (5RO 1-HEO7240 NEUA) from the National Institutes of Health to Professor Vitums, by funds for biological and medical research made available by State of Washington Initiative Measure No. 171 to Professor Vitums; by a Research Career Development Award (5K3 AM-18, 370) from the National Institutes of Arthritis and Metabolic Diseases to Professor King; and by a research grant (5RO 1 NB 06 187) from the National Institutes of Health to Professor Farner. The senior author is greatful to Professor Dr. Hideo Murai and Doctor Yasukuni Watanabe, Department of Animal Science, Shinshu University,Ina,Japan, for their cooperation and support in this investigation. We wish to thank Mrs. Sumiko Sumida for technical assistance, and Miss Kathleen Reinhardt for the preparation of the drawings.     

Lectin-labeled membrane is transferred to the Golgi complex in mouse pituitary cells in vivo

Labeling of the Golgi complex with the lectin conjugate wheat germ agglutinin-horseradish peroxidase (WGA-HRP), which binds to cell surface membrane and enters cells by adsorptive endocytosis, was analyzed in secretory cells of the anterior, intermediate, and posterior lobes of mouse pituitary gland in vivo. WGA-HRP was administered intravenously or by ventriculo-cisternal perfusion to control and salt-stressed mice; post-injection survival times were 30 min-24 hr. Peroxidase reaction product was identified within the extracellular clefts of anterior and posterior pituitary lobes through 24 hr but was absent in intermediate lobe. Endocytic vesicles, spherical endosomes, tubules, dense and multivesicular bodies, the trans-most saccule of the Golgi complex, and dense-core secretory granules attached or unattached to the trans Golgi saccule were peroxidase-positive in the different types of anterior pituitary cells and in perikarya of supraoptico-neurohypophyseal neurons; endoplasmic reticulum and the cis and intermediate Golgi saccules in the same cell types were consistently devoid of peroxidase reaction product. Dense-core secretory granules derived from cis and intermediate Golgi saccules in salt-stressed supraoptic perikarya likewise failed to exhibit peroxidase reaction product. The results suggest that in secretory cells of anterior and posterior pituitary lobes, WGA-HRP, initially internalized with cell surface membrane, is eventually conveyed to the trans-most Golgi saccule, in which the lectin conjugate and associated membrane are packaged in dense-core secretory granules for export and potential exocytosis of the tracer. Endoplasmic reticulum and the cis and intermediate Golgi saccules appear not to be involved in the endocytic/exocytic pathways of pituitary cells exposed to WGA-HRP.     

Ultrastructural characteristics of the differentiation of glandular cells in the anterior pituitary gland of collared lemmings (Dicrostonyx) from the Vrangel Island

In 15.5-, 16-, 17.5-, 18.5-, 20-day-old fetuses and newborn lemmings, the ultrastructure of the anterior lobe of the hypophysis has been studied. In the parenchyma of the 15.5-day-old fetus gland, some cells containing secretory material are revealed, but they are difficult for identification, since differentiation of their organellas is not completed and polymorphism of their granules is strongly manifested. On the 17.5th day of the prenatal development TTH-, ACTH-, LH-, STH-secreting adenocytes are detected; they include secretory granules having diameters 70--100 nm, 80--150 nm, 60--140 nm and 200--400 nm, respectively. On the 18.5th day FSH-secreting cells are identified. Lactotropocytes appear on the 20th day. The anterior lobe of the hypophysis of 17.5--18.5-day-old fetuses are characterized by a high level of the secretory activity which decreases to some extent before birth. Ultrastructure of the cellular organellas in the newborn hypophysis is similar to that in a mature animal, nevertheless, the diameter of the secretory granules does not reach the size of these elements in mature individuals.     

Ultrastructure of neurosecretory cells in the pars intercerebralis of Rhodnius prolixus (Hemiptera).

Six neuron types are distinguished in the pars intercerebralis of the starved fifth instar of Rhodnius prolixus. All neuron types contain electron dense secretory granules derived from Golgi complexes which are of characteristic size and morphology in each type. The neuron types are not thought to represent stages in a secretory cycle. The variety of neuron types described is related to that revealed by staining sections of the same cells with paraldehyde fuchsin. Active synthesis of neurosecretory granules continues throughout starvation and the lysosomal system appears to be involved in the continual degradation of secretory granules. Some of the variations in granule morphology observed may be a consequence of granule fusion and the importance of cytoplasmic events in the development of neurosecretory granules is discussed.