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     

Histology of the pituitary gland of the grey mullet, Mugil cephalus L.

The present communication deals with a histological study of the pituitary gland of the teleost fish Mugil cephalus , found in the estuarine waters of Cochin area. Six different cell types were identified in the pituitary gland on the basis of their grouping, distribution and staining properties. The prolactin and the TSH cells (thyroid stimulating hormone producing cells or thyrotrops) were identified in the rostral pars distalis and the ACTH cells (adrenocorticotropic hormone producing cells or corticotrops) in the interphase between the neurohypophysis and the rostral pars distalis. The STH cells (somatotropic hormone producing cells or somatotrops) and the gonadotropic cells were distinguished in the proximal pars distalis and the MSH cells (melanin stimulating hormone producing cells or melanotrops) in the pars intermedia.     

Immunocytochemical and ultrastructural correlates of the pineal inhibition of prolactin cell activity in blind-anosmic female rats

Summary The effects of the pineal gland on the light microscopic-immunocytochemical and ultrastructural appearance of pituitary mammotrophs were studied in female rats eight weeks after prepubertal blinding and olfactory bulbectomy.Blinding and anosmia resulted in a marked decrease in the size of the pars distalis concomitant with a reduction in the apparent number and size of PRL cells as compared with intact animals. Ultrastructurally, these cells appeared much less active than those of intact rats. The small and angular-shaped mammotrophs of blind-anosmic rats characteristically exhibited scant arrays of rough endoplasmic reticulum, small Golgi complexes with few immature secretory granules, few mature secretory granules and rare exocytosis patterns.Pinealectomy tended to reverse the effects of blinding and anosmia on pars distalis size and PRL cell size, apparent number and ultrastructure. In fact, the mammotrophs of blind-anosmic-pinealectomized rats were quite similar in ultrastructural appearance to those of intact rats.From these data we conclude that the pineal causes mammotroph hypotrophy and hypoplasia in blind-anosmic female rats.Supported by USPHS Biomedical Research Support Grant # RR 05675. The authors thank Dr. Bruce A. Richardson for his kind help with the immunocytochemistry     

大鼠实验性腺胃癌形成过程中脑垂体远侧部ACTH细胞的免疫组织化学观察

用断乳雄性 Wistar大鼠 15只 ,分为实验性胃癌组和盐水对照组。实验性胃癌组大鼠用 MNNG灌胃 ,流式细胞仪测定有异倍体 (10 0 % )后 ,断头处死动物 ,快速取出脑垂体。用免疫组织化学、图像分析和形态计量方法观察大鼠实验性腺胃癌发生过程中脑垂体远侧部 ACTH细胞的变化。结果表明 ,实验性胃癌组大鼠脑垂体远侧部 ACTH阳性细胞的面数密度增大 ,反应增强 ,与对照组相比有显著性差异 (P<0 .0 1,P<0 .0 5 )。结果提示 ,在大鼠实验性腺胃癌发生过程中 ,脑垂体远侧部 ACTH细胞可能参与了机体抗肿瘤的内分泌调节。     

Increased serum prolactin levels mediate the suppressive effects of ectopic pituitary grafts on copulatory behavior in male rats

To determine if deficits in sexual activity observed in pituitary-grafted male rats are due to elevated serum prolactin (PRL) levels found in these animals, the effects of whole pituitary grafts, pars distalis grafts, and ovine (o) PRL treatment on male copulatory behavior were compared. Adult sexually experienced CDF male rats were given four whole pituitary grafts, four pars distalis grafts, or were sham operated. Both groups of grafted animals exhibited suppressed copulatory behavior patterns when tested 18 days after pituitary transplantation. Animals given whole pituitary grafts had significantly longer latencies to mount (P less than 0.05) and to intromit (P less than 0.01) than did the sham-operated controls, while the animals given anterior pituitary grafts differed from the sham-operated controls in latencies to mount (P less than 0.05) and to intromit (P less than 0.01), as well as in the number of intromissions (P less than 0.05). Prolactin-injected animals had significantly reduced intromission rates (P less than 0.01) and significantly increased latencies to mount (P less than 0.05) and to intromit (P less than 0.01) when compared to vehicle-injected controls. Furthermore, the time course of behavioral suppression was similar in oPRL-treated animals to that observed in pars distalis-grafted males, with both groups showing the onset of deficits in sexual activity within 8 to 9 days from the induction of the hyperprolactinemic state. The similarity in pattern and time to onset of behavioral suppression in pituitary-grafted and oPRL-treated animals suggests that behavioral deficits observed in animals with pituitary grafts result from chronic elevation of serum PRL levels.     

Immunocytochemistry of somatotrophs,gonadotrophs, prolactin and adrenocorticotropin cells in larval sea bream (Sparus auratus) pituitaries

Summary The chronological appearance of endocrine cells in the pituitary of sea-bream (Sparus auratus) larvae was studied using antisera against salmon prolactin, trout growth hormone, salmon gonadotropin and N-terminal human adrenocorticotropin. The larval pituitary (1–12 days after hatching) was oval in shape and was composed of a dense mass of cells with few neurohypophysial fibres. By 60 days after hatching it began to resemble the adult and was divisible into a distinct rostral pars distalis containing prolactin and adrenocorticotropin cells; a proximal pars distalis containing somatotrophs and gonadotrophs and a pars intermedia. Cells immunoreactive with antisera against growth hormone were observed immediately after hatching (2 days post-fertilization). Weakly staining prolactin cells were observed 2 days later in the region corresponding to the rostral pars distalis. Cells immunoreactive with anti-gonadotropin and anti-adrenocorticotropin sera were observed in the pituitary 6 and 8 days after hatching, respectively. All the cell-types studied were immunoreactive from the time they were first identified until the final samples 90 days after hatching.     

乌龟脑垂体显微及其腺垂体超微结构的研究

乌龟脑垂体由柄形神经垂体和椭圆形腺垂体两部分组成,神经垂体位于腺垂体后部上方呈背腹型排列。神经垂体中神经叶不发达,腺垂体分为远侧部和中间部,特殊空泡结构成为垂体门脉系统的特征。远侧部细胞分为嗜酸性细胞、嗜碱性细胞和嫌色细胞3种。通过透射电镜观察,腺垂体远侧部主要有5种分泌激素细胞:即生长激素(GH)分泌细胞、催乳激素(PRL)分泌细胞、促甲状腺激素(TSH)分泌细胞、促肾上腺皮质激素(ACTH)分泌细胞、促性腺激素(GTH)分泌细胞和非分泌类型滤泡-星形细胞(FS)。生长激素分泌细胞核大、分泌颗粒少的特征成为乌龟与其他动物最大的区别,可能与乌龟具有生长慢、寿命长的生物学特性有关。       

乌脑龟垂体显微及其腺垂体超微结构的研究

乌龟脑垂体由柄形神经垂体和椭圆形腺垂体两部分组成,神经垂体位于腺垂体后部上方呈背腹型排列。神经垂体中神经叶不发达,腺垂体分为远侧部和中间部,特殊空泡结构成为垂体门脉系统的特征。远侧部细胞分为嗜酸性细胞、嗜碱性细胞和嫌色细胞3种。通过透射电镜观察,腺垂体远侧部主要有5种分泌激素细胞：即生长激素（GH）分泌细胞、催乳激素（PRL）分泌细胞、促甲状腺激素（TSH）分泌细胞、促肾上腺皮质激素（ACTH）分泌细胞、促性腺激素（GTH）分泌细胞和非分泌类型滤泡．星形细胞（Fs）。生长激素分泌细胞核大、分泌颗粒少的特征成为乌龟与其他动物最大的区别,可能与乌龟具有生长慢、寿命长的生物学特性有关。     

Cytological changes in hypophysis of immatur rats after administration of anti-LH antibodies (author's transl)]

In the present work, cytological changes produced in rats' pars distalis were studied by administration of anti-LH antibodies. The anti-LH sera was obtained by active immunization of adult rabbits using bovine LH hormone with Freund's complete adjuvant. Newborn albino Wistar male and female rats were daily and subcutaneously inoculated with 0.1 ml. to 0.8 ml of anti-LH sera from the first 24 hr. on during 5 weeks. For controls, a similar schedule of inoculations with normal rabbit serum was used. At weekly intervals during the treatment, 5 lots of rats were sacrificed and the rest after a recovery period of 95 days. At the end of the 2nd week of immunization, both male and female animals showed degranulated gonadotroph cells in the central part of the pars distalis; at the 3rd week these cells were hypertrophied and presented developed Golgi complex. At the 4th week, the first large vacuoles of "castration" were seen in the gonadotroph cells of male pars distalis, besides in the females the degranulation of gonadotrophs continued. During the last week of treatment, the gonadotrophs of the male animals presented a highly dilated Golgi complex, more "castration-like cells" and numerous mitosis figures. The gonadotrophs of females pars distalis presented degranulation, but not vacuoles. After the recovery period the gonadotrophs of male rats were similar to those of the control hypophisis and did not show castration vacuoles in their cytoplasm. In female rats, the gonadotrophic cells showed "castration-like vacuoles" and a raised number of mitosis cells compared to control pituitaries. The significance of these findings will be discussed.     

Pituitary gland of the garter snake (Thamnophis): Morphology and immunocytochemistry

The three-dimensional gross morphology of the pituitary gland of the garter snake (Thamnophis sirtalis) is presented. Hormone-producing cells of the pars distalis were localized immunocytochemically. Corticotropes and lactotropes occur in the anterior two-thirds of the gland; corticotropes are especially numerous in the area of the pars distalis nearest the median eminence, and lactotropes are most abundant medially. Somatotropes are restricted to the posterior one-third of the pars distalis. Gonadotropes and thyrotropes are scattered throughout the pars distalis and in favorable sections form a network of cells enclosing clusters of peptide-secreting cells.     

Seasonal changes in the pituitary gland of the feral hawaiian mongoose (Herpestes auropunctatus)

The Hawaiian mongoose (Herpestes auropunctatus) is a seasonally breeding mammal whose pituitary gland resembles that of other Viverridae. Certain features, such as a prominent pars tuberalis interna and a double-layered pars intermedia forming a cup for the neurohypophysis, are unique. With the light microscope, five different cell types can be recognized in the pars distalis after staining with periodic acid-Schiff (PAS)-orange G. Two types of acidophils are seen, a small yellow-staining cell and a large angular orange cell. Two basophilic cells are also seen, one with fine PAS-positive cytoplasmic granules and the other with coarse PAS-positive granules in the cytoplasm. The last cell type seen is the chromophobe. Differential cell counts indicated an altered distribution of chromophils in the ventral pars distalis of the female mongoose with changing season and reproductive status, but the most striking change was a decreased percentage of basophils in the pars distalis during the nonbreeding season.     

Uptake and retention of sex steroids by the baboon pituitary gland--evidence of sexual dimorphism with respect to dihydrotestosterone

The localization and identification of the target cells for estradiol (E2) and dihydrotestosterone (DHT) were studied in the pituitary gland of male and female baboons (Papio cynocephalus) using autoradiography and autoradiography combined with immunocytochemistry. The largest number of cells which sequestered 3H-E2 were located in the pars distalis of both the males and females. Immunocytochemistry revealed that these target cells were predominantly gonadotrophs and mammotrophs, albeit a few somatotrophs and thyrotrophs were also labeled. A distinct sexual dimorphism was not observed with respect to the uptake and retention of the tritiated estrogen but was clearly evident in those animals given 3H-DHT. Significantly more cells in all three lobes of the male baboon, the partes distalis, intermedia and nervosa, concentrated the radiolabeled androgen as compared to that found in the females. Regardless of sex, however, the gonadotrophs were the major cell type in the pars distalis which was labeled. A small population of mammotrophs, somatotrophs and thyrotrophs also retained 3H-DHT. In both males and females, the pars nervosa was the portion of the hypophysis most sensitive to 3H-DHT. These findings indicate that all three lobes of the hypophysis of the male and female baboon contain target cells for E2 and DHT and that the propensity for DHT uptake is greater in males than in females.     

Immunocytochemistry of a “private” luteinizing-hormone-releasing hormone system in the pituitary

Summary Immunocytochemistry of paraffin sections of Bouin-fixed rat pituitaries with antiserum to luteinizinghormone-releasing hormone (LHRH) revealed two types of cells. Type I cells exhibit granular staining throughout their cytoplasm. The immunoreactivity of type II cells is confined to a much smaller area of the cytoplasm. Type I cells are located in the ventral margin of the pars intermedia, the region between the pars intermedia and the pars distalis, and the pars distalis adjacent to this region. Type II cells have a broader distribution in the pars distalis, but tend to concentrate in the region of the pars distalis near the pars intermedia. Type I cells are distinct from gonadotropes. Type II cells appear to comprise a subgroup of gonadotropes. Staining in type I, but not type II, cells in pituitary explants, maintained in serum-free media for seven days, is as intense as that in normal pituitary tissue. The data suggest that the type I cells are producing an intrinsic LHRH-like material and may be responsible, in part, for the regulation of some gonadotropes.Supported by NIH grants HD12932, NS15843 and NS15809 (LAS), National Science Foundation grant BNS 82-05643 (LAS), and a grant from the Phillippe Foundation (JYL)     

The Cell Types in the Adenohypophysis of the Marine Teleost Crenilabrus melops

Sagittal sections from pituitaries of winter and spring fish were stained with a number of methods including alcian blue-PAS-orange G, Herlant's tetrachrome, Luxol fast blue-PAS-orange G and lead haematoxylin. By these methods seven different endocrine cell types were readily demonstrated in the adenohypophysis. In the rostral pars distalis (RPD), there were conspicuous vacuoles (intra- and possibly extracellular) in the prolactin cell zone of all animals. The ACTH cells in the RPD were very numerous. In the proximal pars distalis (PPD), the growth hormone cells were arranged in coiling bands that were predominantly dorsal and juxtaposed to neurohypophysis. Two types of basophils were readily distinguished: both were predominantly ventral in the proximal pars distalis, though the type I basophil also formed a constant group in the dorsal region of the pituitary, between the PPD and the RPD. The pars intermedia contained two cell types — a lead haematoxylin-positive cell and a PAS-positive cell. The pituitary morphology is compared with that of other teleosts.     

Endorphin in rat pituitary glands: its distribution within the gland, and age related changes in gland content in male and female rats

Endorphins were extracted with glacial acetic acid:acetone from freshly microdissected single lobes of rat pituitary and assayed for inhibition of ³H-etorphine stereospecific binding to rat brain opiate receptors. Highest tissue concentrations of endorphin were found in pars intermedia and pars distalis, with minimal activity observed in pars nervosa. In addition, mature female rats exhibited a significantly higher anterior lobe endorphin content than males, reflecting the larger gland weight, although no significant differences were found between neurointermediate lobe endorphin content of either sex. Pituitary endorphin content increased substantially with age, rising rapidly between the fifth and tenth week after birth. This elevation was still apparent at twenty-five weeks. These studies emphasize the importance of employing rats of closely similar age and weight in any experiments designed to examine pituitary endorphin function, and suggest that ontological development of pituitary endorphin may proceed in a manner similar to that of other pituitary hormones (e.g., MSH) derived from the same precursor peptide.     

Immunohistochemical identification of cells in the pars distalis of the pituitary of the lizard Anolis carolinensis

Immunocharacteristics of the pars distalis cells of the pituitary of the male lizard A. carolinensis are determined by employing the immunoperoxidase technique with antisera to mammalian pituitary hormones. On the basis of their immunoreactivity, 5 different cell types with characteristic anatomical distribution are recognized. ACTH cells are found in the rostral half of the pars distalis, and PRL cells in the rostral two thirds of the pars distalis. GH and TSH cells are located in the caudal half of the pars distalis. GTH cells are distributed throughout the gland. When consecutive sections are stained with antiserum to ovine FSH or its beta-subunit and to ovine LH, the same cells show immunoreactivity to all the three antisera. None of the GTH cells show positive immunoreactivity to ovine beta-LH antiserum. The results suggest the existence of one gonadotropic cell type in the pituitary of this lizard.     

Immunohistochemically detected ontogeny of prolactin and growth hormone cells in the African catfish Clarias gariepinus

The chronological appearance of selected endocrine cells in the pituitary of African catfish Clarias gariepinus (Burchell, 1822) was studied morphologically, histologically and immunohistochemically by using antisera raised against catfish growth hormone (cgGH) and recombinant tilapia prolactin I (tPRL). cgGH- and tPRL-like immunoreactive cells were visible from day 1 post fertilisation (hatching) throughout the juvenile and the adult stage. From 1 to 90 days after hatching, the larval pituitary is oval in shape with a distinctly shaped rostral pars distalis, proximal pars distalis and pars intermedia. From day 120 onwards allometric growth of the rostral and proximal pars distalis extended the prolactin and growth hormone cells anteriorily and posteriorily, respectively. Size and activity of the prolactin and growth hormone cells, measured by the ratio of cell surface to nuclear surface remained constant until day 40 and showed a growth spurt thereafter. Growth hormone content, measured with a catfish-specific radio-immunoassay from hatching until 60 h post hatching, increased exponentially between 30 and 60 h.     

A histological study of the hypophysial-gonadal system during sexual maturation and spawning in the milkfish, Chanos chanos (Forskal)

The pituitary gland of the milkfish, Chanos chanos , was studied at different stages of sexual maturation and spawning. Consecutive median sagittal sections were treated with a range of stains to demonstrate the different cell types and regions. The milkfish pituitary consists of a neural component, the neurohypophysis, and an epithelial component, the adenohypophysis, which in turn consists of three regions: the rostral pars distalis (RPD), proximal pars distalis (PPD), and pars intermedia (PI). However, unlike most teleosts, the pituitary gland of the milkfish is encased in a bony chamber, has dorsal and ventral lobes and extends anteriorly from its point of origin at the base of the brain. PAS (+) basophils are found in all regions of the adenohypophysis, but mostly in the proximal pars distalis. These cells undergo hypertrophy and hyperplasia during sexual maturation, shrinkage and degranulation during spawning.     

Life span changes in the presence of alpha-melanocyte-stimulating-hormone-containing cells in the human pituitary.

Since recent circumstantial evidence has suggested possible functions of alpha-MSH in intrauterine growth and labour, the presence of this hormone in the human pituitary was determined by means of the indirect immunofluorescence procedure during development and adulthood. Cross reaction of the antibodies with other peptides was measured after which they were purified by solid phase absorption. Experiments on the rat pituitary showed that staining of alpha-MSH- and ACTH-containing cells could be obtained well until 48 h after death. In the pars distalis the ability of ACTH-containing cells to take up stain increased during the period of post-mortem storage. In the youngest human fetus studied (15 weeks) only alpha-MSH-containing cells were found in the pars intermedia and no ACTH-containing cells were observed. In the other fetal pituitaries a distinct pars intermedia containing more alpha-MSH cells than ACTH cells was found. In the pars distalis of the fetuses more ACTH- than alpha-MSH-containing cells were observed. From birth to 19 years, progressively fewer alpha-MSH containing cells could be detected in the 'zona intermedia' and pars distalis, while in adults only a few such cells were found in either area. Irrespective of age, sex, cause of death or therapy, alpha-MSH-containing cells were found in all pituitaries throughout life. The number of ACTH containing cells gradually increased in the zona intermedia and pars distalis and reached a high adult level in the latter structure. In the pituitaries of seven anencephalics, no alpha-MSH-containing cells were present. The presence of alpha-MSH in the fetal pars intermedia, the change in the ratio of the alpha-MSH/ACTH cells during the course of development, and the absence of alpha-MSH in anencephaly all support the possibility that human fetal pituitary alpha-MSH is involved in both intrauterine growth and fetal adrenal function and thus also in parturition.     

Peptides with NH2-terminal tryptophan in adrenocorticotrophic hormone and melanocyte-stimulating hormone granules of adenohypophysis.

Fluorescence microscopy has demonstrated formaldehyde-ozone-induced fluorescence in the pars intermedia cells (melanocyte-stimulating hormone cells) and in certain cells of the pars distalis of the mammalian pituitary. From histochemical and chemical evidence the fluorescence is believed to reflect the presence of peptides with NH2-terminal tryptophan. In the pars distalis of hamster, cat and pig pituitary, the cells that exhibit formaldehyde-ozone-induced fluorescence have now been identified as adrenocorticotrophic hormone (ACTH) cells by immunohistochemistry. Granules from pig pituitaries were purified by passage through a succession of Millipore filters followed by centrifugation on a continuous sucrose gradient. Two granular fractions were identified by electron microscopy and found to contain high concentrations of peptides with NH2-terminal tryptophan as well as high ACTH bioactivity. These fractions, when pelleted and analyzed histochemically, displayed formaldehyde-ozone-induced fluorescence and ACTH-like immunoreactivity.