Somatotrophs and lactotrophs in the anterior pituitary of fetal and neonatal rats

Summary The ultrastructure of immunoreactive somatotrophs and lactotrophs in pituitaries of fetal rats at 19, 20 and 21 days of gestation and on the day of birth was studied. Somatotrophs, first detectable at 19 days of gestation, undergo only minor modifications before reaching the structure described for adults. In particular there is an increase in the endoplasmic reticulum and Golgi apparatus. Lactotrophs, first identifiable in newborn rats, are very different in ultrastructure from adult cells, because the secretory granules are generally small, but variable in shape and size, and the Golgi complex is prominent.     

Fate of degenerating lactotrophs in rat pituitary gland after interruption of lactation: a histochemical and immunocytochemical study

Summary In the pituitary gland of pregnant and lactating rats a striking proliferation of lactotrophs occurs to meet the increased demands for prolactin. Following interruption of lactation the redundant lactotrophs undergo a massive degeneration until pre-pregnant proportions are re-established. Immunocytochemical detection of prolactin allows the recognition of degenerating lactotrophs until advanced stages of degeneration and leads to the conclusion that this process is autolytic in nature. Histochemistry of acid phosphatase reveals a remarkable accumulation of this enzyme in Golgi cisternae and lysosomes. At later stages of degeneration the acid phosphatase spreads throughout the entire cell. The presence of increased numbers of necrotic cells appears to activate phagocytosis of stellate cells and, to a lesser extent, of follicular cells. Stellate cells responsible for the secondary processing of cell residues are isolated cells characterized by a prominent oval nucleus and an electron-lucent cytoplasm with scarce organelles and extensive cytoplasmic processes. They appear as scavenger cells engulfing cell remnants and debris. Immunocytochemistry of S-100 protein discloses differential staining of two types of cell, one forming clusters of 2–4 cells with faint immunoreactivity, while the other type consists of isolated cells with a stellate profile and stronger labelling to S-100 protein.     

Innervation of the canine mammary gland: an immunohistochemical study

The distribution of peptidergic nerves in canine mammary tissues was studied by immunohistochemical techniques. In addition, the general and the noradrenergic innervations were demonstrated using protein gene product 9.5 and tyrosine hydroxylase immunoreactivities as markers, respectively. Tissue specimens from the caudal mammary glands were obtained from adult, non-lactating, female dogs. The overall innervation of the mammary gland tissue was sparse and primarily associated with the arterial vasculature. Nerve fibres positive for protein gene product 9.5 were rarely found in the secretory parenchyma. The nipple was not richly innervated, although it displayed a greater amount of nerve fibres than the mammary parenchyma. Nerve fibres supplying nonvascular structures of the nipple expressed immunoreactivity for the sensory neuropeptides calcitonin gene-related peptide, substance P and neuropeptide K, but not for vasoactive intestinal peptide, peptide histidine isoleucine and C-flanking peptide of neuropeptide Y. Somatostatin immunoreactivity was not detected in mammary gland tissue. Our results indicate that the innervation of the canine mammary gland is mainly affiliated with the vasculature and comprises peptidergic nerves which may be involved in the regulation of local blood flow. The presence of sensory neuropeptides in nerves supplying the mammary nipple suggest that these peptides may play a role in the afferent pathway of the milk ejection reflex.     

Interrelationships between the hypothalamus, pituitary gland, ovary, adrenal gland, and the open period for LH release in the hen (Gallus domesticus)

The asynchronous ovulatory cycle of the hen is believed to be the consequence of two interacting systems, one of which is circadian and regulates the timing of the preovulatory LH surge. In support of this proposition, the open period for LH release was shown to oscillate with the same periodicity as the photoschedule when the hens were exposed to 14 L:7 D, 14 L:10 D, and 14 L:14 D. In addition, it was demonstrated that follicular maturation is not affected by or synchronized with the photoperiod. The physiological system that transduces the light/dark cycle into an open period for LH release has not been identified although circumstantial evidence supports the idea that the adrenal gland plays a role in this function. This evidence includes the anatomical juxtaposition of the left ovary and adrenal gland, innervation of steroid-producing cells within the follicle by nerve tracts passing through the adrenal glands, the ability of injections of metyrapone to alter the timing of preovulatory LH release, the ability of injections of corticosterone to induce ovulation when a mature follicle is present in the ovary, and the ability of dexamethasone or infusions of corticosterone to block ovulation. Recently we have also shown that infusions of corticosterone will block the gonadotropic effect of PMSG, will inhibit the photoperiodic response, and do not affect the release of LH in response to injections of GnRH. The addition of corticosterone to incubations of dispersed granulosa cells does not affect their response to LH. These data suggest that corticosterone may modulate the responsiveness of the hypothalamus to tropic stimuli and demonstrate that exposure to corticosterone can alter the responsiveness of some ovarian tissues to gonadotropins.     

Detection of gonadotrophic hormones on isolated granulosa cells of the domestic hen, Gallus domesticus, by an immunohistochemical method

Two immunohistochemical methods were used to detect the presence of luteinizing hormone (LH) on the cells of chicken granulosa. Using a peroxidase-labelled anti-rabbit serum together with anti-chicken LH serum raised in rabbits, a strong positive response was obtained with granulosa cells from small and large pre-ovulatory follicles obtained from the mid-cycle. Similarly, by using an available antiserum to human follicle stimulating hormone (FSH), a slightly weaker response was obtained with cells from both large and small follicles. After incubating cells with ovine LH, ovine FSH and ovine prolactin, there was no detectable difference with the method used in reaction with their respective antisera to cells which had received no incubation, implying that chicken gonadotrophins were displaced only partially from their receptors by mammalian gonadotrophic hormones. Pre-incubation of the cells with human chorionic gonadotrophin gave negative results with anti-hCG serum. Using a fluorescent-labelled antibody method, similar results were obtained except that the distribution of the receptors on the granulosa cell for LH or FSH appeared to be different. With the LH, the fluorescence formed a halo around the cell in contrast to the overall fluorescence with FSH.     

Brain and pituitary beta-endorphin levels at different developmental stages of the rat

beta-Endorphin-like immunoreactivity in whole brains of Sprague-Dawley rat fetuses of different gestational ages was measured by radioimmunoassay and found to increase throughout the gestational period studied. The immunoreactivity in various brain parts (forebrain, midbrain, hindbrain, hypothalamus and pituitary) of late prenatal, early postnatal, young mature and retired breeder rats was also determined. In all the brain parts studied, a maximum in the content and concentration of beta-endorphin-like immunoreactivity was attained when the rats were about 3-4 months old.     

Detection of gonadotrophic hormones on isolated thecal interstitial cells of the domestic hen, Gallus domesticus, by an immunohistochemical method

Summary An immunohistochemical method was used to demonstrate the presence of gonadotrophins in isolated ovarian interstitial cells. The cells were obtained by collagenase digestion of large ovarian follicles after removal of the yolk and the granulosa layer. Using a peroxidase-labelled anti-rabbit serum with anti-chicken follicle stimulating hormone (FSH) serum raised in rabbits, a strong positive reaction was obtained. Anti-human FSH serum also produced a positive result but the reaction was weaker. There was no apparent difference in the staining reaction of cells which had been preincubated with ovine FSH serum. Treatment with anti-ovine luteinizing hormone (LH) resulted in a faintly positive reaction.The viability of the cells was tested by the Trypan Blue method and they were identified as steroid-producing cells by the histochemical demonstration of their 3-hydroxysteroid dehydrogenase activity.     

Melatonin-like immunoreactivity in the pineal gland of the cow: an immunohistochemical study

With a view to checking the presence of melatonin in the pineal gland of the cow, in the present work we used six adult animals, ranging in age from one to six years, which were sacrificed at dawn. Sections of 6 micro m thickness of Bouin-fixed and paraffin-embedded pineal glands were incubated in an anti-melatonin serum, which was provided by the Institute for Molecular and Cellular Recognition, Gunma University, Maebshi, Japan. After incubation and successive washings in PBS, some of the sections were treated with the avidin-biotin-peroxidase complex (ABC) technique using antisera from Sigma, and developed with the method of Graham and Karnovsky (which employs 3,3'-diaminobenzidine and H2O2 as developer). Other sections were incubated in a goat-anti-rabbit IgG (H+L) bound to fluorochrome Cy5 for immunofluorescence studies. An intense reaction for melatonin was observed in the cytoplasm but not in the nucleus of melatonin secreting pinealocytes located in peripheral and intermediate zones of the pineal gland. Immunoabsorption of the antimelatonin primary antibody with melatonin at a dilution of 10 mM per 0.1 ml of serum prevented the reaction, as happened when any of the antisera used in the procedure were used. Immunoabsorption of anti-melatonin serum with different amounts of bovine albumin (ranging between 1/5 to 1/50) failed to inhibit the immunoreactivity. When a bovine anti-albumin antibody was employed, working with the above methods, no immunoreaction was detected. Our data suggest that the pinealocytes of cows sacrificed at dawn contain immunoreactive melatonin.     

Comparative cytological study of Phasianus colchicus,Meleagris gallopavo,and Gallus domesticus

Summary Since viable intergeneric hybrids between the chicken (Gallus domesticus) and the pheasant (Phasianus colchicus) have been reported, as well as interfamilial hybrids between the chicken and the turkey (Meleagris gallopavo), the chromosome complements of the pheasant and the turkey were compared with that of the chicken. In these three species belonging to the order Galli, the Z-chromosomes appeared to be identical, while the autosomal complements of the pheasant and the turkey differed radically from that of the chicken. It was noted with some surprise that the pheasant of the family Phasianidae and the turkey of the family Meleagridae have very similar chromosome complements, at least so far as gross morphology of somatic metaphase chromosomes is concerned.This work was supported in part by grant C-5138 from the National Cancer Institute, U.S. Public Health Service, and grant C-17601 from the National Science Foundation.The authors gratefully acknowledge the generosity of Rea's Game Birds, Paramount, California, who supplied the pheasant chicks, and the McPherin Hatcheries, Sunnymead, California, who furnished the turkey chicks. The authors also appreciate the editorial assistance of'Patricia A. Ray.     

Free amino acids in claw muscle and haemolymph from Australian freshwater crayfish at different stages of the moult cycle

Amino acids were measured in claw muscle and haemolymph in the freshwater decapod crustacean, Cherax destructor, at different stages of the moult cycle. The total pool of amino acids in muscles from animals in intermoult (97+/-13 mmol kg(-1) muscle), premoult (80+/-20 mmol kg(-1)) and postmoult (97+/-19 mmol kg(-1)) were not significantly different. Despite the relatively stable total pool of amino acids, there were changes in the concentrations of alanine, glutamine and proline over the moult cycle. Compared to intermoult, claw muscles from animals in premoult had a lower concentration of proline, and animals in postmoult had higher concentrations of alanine and glutamine, but lower concentrations of proline. Concentrations of alanine and glutamine in claw muscle of animals in postmoult were higher and proline concentrations lower than in the same animals during the premoult stage. The concentration of proline in haemolymph was lower in animals in premoult and postmoult compared to intermoult. The total amino acid pool in the claw muscle of Cherax destructor did not change significantly over the moult which is distinctly different to the changes in amino acids reported in the claw muscles of marine decapod crustaceans.