Nuclear uptake and retention of androgen by the pituitary gland of the hamster and the rat

Summary Castrated adult male hamsters and castrated adult female rats were injected with either 0.2 g (hamsters) or 0.5 g (rats) of ³H-dihydrotestosterone (107 Ci/mmole)/100 grams body weight and killed 11/2 h later. The pituitary glands were removed and processed for both autoradiography and immunocytochemistry (hamster) or only autoradiography (rats). Localization of the androgen was found in 10–15% of the cells of the pars distalis in both species. Only cells that stained for luteinizing hormone (LH) in the hamster's pars distalis concentrated the androgen. Also cells in both the pars intermedia and pars nervosa (1–5%) concentrated the androgen in both species. Although the number of cells that concentrated the androgen in the pars intermedia and pars nervosa was small, this finding may be related to recent physiologic data that suggest that the gonadal steroids may play a role in regulating water retention and natriuresis.This study was supported by USPHS Research Career Development Award KO4NS0000164 (P.J. Sheridan) and USPHS Grants No. 1 RO1 NS12933, P30 HD10202 and HD 10914     

Ultrastructure of STH cells in the pars distalis of the hypophysis of mice bearing transplanted hepatomas

Summary An electron microscopic analysis was performed on the pars distalis of the hypophysis of mice bearing transplanted hepatomas. Normal mice served as controls. In animals bearing a fast growing as well as in those bearing a slow growing hepatoma, the STH cells presented changes indicating increased release and synthesis of its secretory product. These changes can be summarized as follows: 1) Increased number of granuledepleted STH cells which were very irregular in size and shape. 2) Very irregular and sometimes bizarre nuclei. 3) Plasma membranes irregular and convoluted. 4) Increased quantity of endoplasmic reticulum which appeared sometimes dilated and contained material of varying electron density. 5) Abundant, irregular and frequently ramified mitochondria. 6) Hypertrophic Golgi system with increased number of microvesicles. 7) Increased number of large lysosomes of different type. 8) Increased number of fat droplets.Work carried out with the financial assistance of grants of the Consejo Nacional de Investigaciones Científicas y Técnicas de la Republica Argentina and the Comisión de Investigación Científica de la Universidad Nacional de La Plata. Thanks are due to the members of the technical Staff of the Institute for their technical assistance.     

Effects of tumors on the daily mitotic activity of mouse pars intermedia

We have investigated the effects of EA21a and EA34 mammary carcinomas on daily PI cell proliferation in mice. Animals were divided into groups grafted with either EA34 or EA21a carcinomas (and a non-grafted control group). They were all injected intraperitoneally with 2 microg colchicine per g of body weight 4 h before sacrifice and the number of mitoses per 1000 nuclei was calculated. The mitotic index (MI) of pars-intermedia epithelial cells in control animals showed significant temporal variations. However, the MI from mice grafted with EA34 or EA21a carcinomas showed no such variation. There was no difference between the daily MIs of controls and tumor grafted groups. The absence of a 24 h mitotic activity curve in both EA21a and EA34 tumor-bearing animals demonstrates a lower level of synchronization of cells entering mitosis.     

A light and electron microscopic study of the pre- and postnatal development and secretory differentiation of the pars tuberalis of the rat hypophysis

Summary The development of the pars tuberalis was studied in the rat fetus from 13 days of gestation to 6 weeks after birth. After the closure of Rathke's pouch, the pars tuberalis anlage is clearly distinguishable from the anlagen of the partes intermedia and distalis. It comprises the entire basal portion of the adenohypophysial anlage; the limit between the anlagen of the pars tuberalis and the pars distalis is defined by Atwell's recess, i.e. the pathway taken by the hypophysial vessels coming from the vascular plexus of the median eminence.At 14 days the pars tuberalis cells are characterized by the presence of glycogen which persists in the adult. Their secretory differentiation (elaboration of granules with a diameter of 100–120 nm) is obvious at 15 days of gestation. It therefore, clearly precedes that of the other hypophysial cell types. Its functional differentiation takes place well before its adhesion to the primary vascular plexus of the portal system. Cystic formations appear just before birth in the pars tuberalis, much later than those of the pars distalis.These observations on the development of the pars tuberalis, together with previous observations on the adult PT in various species, showing that the specific glandular cells of the pars tuberalis are cytologically different from all known adenohypophysial cell types, seem to indicate a specific endocrine function of this lobe.     

Photoperiodic Variation in CD45-Positive Cells and Cell Proliferation in the Mediobasal Hypothalamus of the Soay Sheep

The Earth's solar orbit induces annual climatic changes challenging to survival. Many animals have evolved to cope with seasonal variability through compensatory annual changes in their physiology and behavior, which involve innate long-term timing and photoperiodic synchronization to anticipate the environmental seasonal cycles. Here we considered the potential involvement of cyclical histogenesis in seasonal timing mechanisms in the sheep. Adult Soay rams were established in three distinctive seasonal states by controlled photoperiod exposure. A first group, representing the condition in late spring (long-photoperiod [LP] group), was taken indoors in May and exposed to 4 wks of 16?h light/day (LP). A second group was exposed to 20 wks of LP to establish a late-summer/long-day refractory condition (LPR group). A third group of animals was brought indoors in August and exposed to 4 wks of LP followed by 4 wks of 8?h light/day (short photoperiod [SP]) to establish an autumn-like condition (SP group). At the end of these regimes, we injected 5-bromo-2-deoxyuridine (BrdU), and animals were killed 24?h or 4 wks later. When BrdU was administered 24?h before death, more BrdU-immunopositive cells were detected in the hilus of the hippocampus in LP compared with SP animals, indicative of a higher proliferation rate. When BrdU was administered 4 wks before death, more BrdU-positive cells were detected in the hippocampus under LP, compared with SP, indicating increased cell survival. These mitotic cells were occasionally seen to adopt a neuronal phenotype in the hippocampus, but not in the hypothalamus. Approximately 10% of BrdU-positive cells in the basal hypothalamus coexpressed the pan-leukocytic marker CD45, and showed morphological features and regional distribution consistent with ameboid microglia. Increased numbers of these cells were detected in the region of the median eminence and tuberoinfundibular sulcus of animals kept in SP compared with LP or LPR. These data suggest that neuroimmune mechanisms may be involved in photoperiod-dependent seasonal remodeling of the adult brain. (Author correspondence: d.hazlerigg@abdn.ac.uk)     

Intercellular communications within the rat anterior pituitary. XVI: Postnatal changes of distribution of S-100 protein positive cells,connexin 43 and LH-RH positive sites in the pars tuberalis of the rat pituitary gland. An immunohistochemical and electron microscopic study

The architecture of luteinizing hormone-releasing hormone (LH-RH) nerve ends and the S-100 protein containing folliculo-stellate cells forming gap junctions in the pars tuberalis is basically important in understanding the regulation of the hormone producing mechanism of anterior pituitary glands. In this study, intact male rats 5–60 days old were prepared for immunohistochemistry and electron microscopy. From immunostained sections, the S-100 containing cells in pars tuberalis were first detected on day 30 and increased in number to day 60; this was parallel to the immunohistochemical staining of gap junction protein, connexin 43. LH-RH positive sites were clearly observed on just behind the optic chiasm and on the root of pituitary stalk on day 30. On day 60, the width of layer increased, while follicles and gap junctions were frequently observed between agranular cells in 10 or more layers of pars tuberalis.     

Mitotic activity of the pars intermedia in the female mouse: age-associated variations in proliferation rate and circadian periodicity

We previously reported daily variations in the mitotic activity of the endocrine cells in the pars intermedia of 21- and 28-day-old male mice. Since cellular proliferation might be affected by factors such as sex and age, we undertook the present experiments to study the mitotic activity of the pars intermedia from 14-, 28-, and 150-day-old female mice. Inbred C3H/S mice, grouped according to age, were housed under standard conditions (12h each of light and dark [LD 12:12]) for periodicity analysis and were killed in lots of 5-11 animals every 4h over a single 24h cycle, with each mouse receiving 2 μg/g of colchicine 4h before decapitation. Pituitaries were excised, extracted, fixed in buffered formaldehyde, embedded in celloidin-paraffin, sectioned at 5 μm, and stained with hematoxylin and eosin. We counted the total number of nuclei to estimate the total number of cells monitored and then calculated the mitotic index (metaphases/1000 nuclei). Differences were analyzed for statistical significance by the Student t test. While the 14-day-old animals manifested no significant changes in mitotic activity during the 24h cycle, the 28- and 150-day-old mice showed higher mitotic indices during the period of darkness. The average mitotic activity over the entire cycle, however, was higher in the two groups of younger animals than in the 150-day-old mice. Moreover, the averages for the 28-day-old females were higher than the corresponding values previously reported by us for male mice of the same age. (Chronobiology International, 17(6), 751-756, 2000)     

Immunocytochemical identification of growth hormote (GH) cells in the pituitary of three anuran species using an antiserum against purified bullforg GH

An antiserum was prepared against the recently purified bullfrog (bf) growth hormone (GH); it was applied to sections of brain and pituitary of three urodele (Ambystoma, Pleurodeles and Cynops) and three anuran (Xenopus, Bufo vulgaris and B. japonicus) species. No immunostaining was obtained in the urodele pituitary, being consistent with the results of immunoblot analysis of the pituitary homogenate. In the three anuran species, strong immunoreactivity was observed in GH cells that were concentrated in the posterodorsal region of the pars distalis. No GH-like immunoreactivity was detectable in the brain of any of the species. A comparison using adjacent sections stained with anti-bf prolactin (PRL) confirmed the anteroventral localization of PRL cells. Colocalization of GH and PRL was not apparent. These data suggest that the molecular structure of amphibian GHs is considerably different between anurans and urodeles. The antiserum used in the present work shows a high species specificity, recognizing only anuran GHs. In contrast anti-bfPRLlabeled PRL cells in all the amphibian species studied in the present work, suggesting that PRLs possess common amino acid sequences recognized by the anti-bfPRL.     

Sur l'existence de cellules de type ≪corticotrope≫ dans la pars intermedia de l'hypophyse du rat

Résumé Nos observations démontrent l'existence de deux types de cellules glandulaires dans la pars intermedia de l'hypophyse du rat: des cellules propres, à M.S.H., dont les grains de sécrétion sont détruits par la fixation osmiée et relativement bien conservés par la fixation au glutaraldéhyde; des cellules à petits grains denses, osmio-résistants, qui s'apparentent morphologiquement aux éléments corticotropes de l'antéhypophyse. L'existence bien connue d'A.C.T.H. ou, plus précisément, d'un facteur A.C.T.H.-like dans le lobe neuro-intermédiaire nous fait considérer comme très vraisemblable la nature corticotrope de ce deuxième type cellulaire de la pars intermedia.

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On the existence of cells of the Corticotrophic type in the intermediate lobe of the rat hypophysisAn electronmicroscopic study

Summary Two types of glandular cells have been shown to occur in the intermediate lobe of the rat hypophysis: 1) M.S.H. producing cells whose secretory granules are destroyed by osmium fixation and relatively well preserved by glutaraldehyde fixation, 2) cells with small dense osmium resistant granules being morphologically similar to the corticotrophic cells of the anterior lobe. The well known occurrence of A.C.T.H. or, more precisely, of an A.C.T.H.-like factor in the neuro-intermediate lobe makes the corticotrophic nature of this second cell type very likely.

Travail dédié à la mémoire de Nicole Granboulan.     

The hypothalamo-hypophysial system of the wild carp,Cyprinus carpio L.

Summary The posterior neurohypophysis (PNH)-pars intermedia complex of the wild and pond carp, Cyprinus carpio L., has been studied by light, fluorescence and electron microscopy. Gomori-positive neurosecretory fibres are abundant in the main trunk of the neurohypophysis as well as its roots penetrating the pars intermedia. Terminals of these fibres are in contact with capillaries of the general circulation and with glandular cells of the pars intermedia. Monoaminergic fibres with a weak green fluorescence, somewhat increasing after injection of nialamide into the pond carp, have largely the same distribution. Three types of neurosecretory fibres and their terminals have been recognized in the PNH-pars intermedia complex. Types-A₁ and -A₂ fibres, containing granules of 140–180 nm and 100–160 nm in diameter respectively, are peptidergic Gomori-positive. Type-A₂ fibres predominate in the PNH. The least frequent monoaminergic type-B fibres have granules of 60–100 nm in diameter. Numerous peptidergic and few monoaminergic neurosecretory terminals make contact with the capillaries located within the roots of the PNH as well as at the border between them and the pars intermedia. Both peptidergic and monoaminergic terminals make direct synaptoid contacts with the gland cells or end close to connective tissue septa, basal lamina or pituicytes. The PAS-positive gland cells and to a lesser degree the leadhaematoxylin-positive gland cells show these relationships with neurosecretory terminals. The question concerning the mode of interaction between peptidergic and monoaminergic structures in the dual control of the gland cells of the pars intermedia of teleosts is discussed.