György Rappay

Pituitary corticotrophs proliferate temporarily after adrenalectomy

SummaryRelationship of corticotroph proliferation answer and survival time after adrenalectomy was examined. Corticotroph proliferation rates were detected by short-term 3H-thymidine radiolabeling, then ACTH immunostaining and autoradiography. Effect of adrenalectomy on corticotroph proliferation rate was examined in vivo and an elevation was demonstrated first on the second postoperative day, increasing on the third-fourth day postoperatively and then decreasing. Effects of different secretagogues on corticotroph proliferation were examined in short-term pituitary monolayer cultures taken from ADX rats. CRF and Forskolin treatment potentiated corticotroph proliferation in cultures taken from adrenalectomized rats, but not in the controls. We suggest that croticotroph proliferation is stimulated via the cAMP-proteinkinase A pathway, while adrenalectomy plays a permissive role.

Growth and fine structure of monolayers derived from adult rat adenohypophyseal cell suspensions

SummaryPrimary monolayer cell cultures of trypsin-dispersed adult rat adenohypophyses are capable of synthetizing deoxyribonucleic acid and are able to divide at least for 3 weeks. Most cells contain secretory granules in the absence of hypothalamic extracts. It seems possible that granules are formed which mature and are released in vitro.

A quantitative approach to trace the corticotrophs in culture after adrenalectomy

SummaryAnterior pituitaries of adrenalectomized and sham operated adult rats were dispersed by trypsin and cultured for 4 and 8 days. Adrenalectomy caused a moderate increase in number of corticotrophs in both zero-time cell suspensions and cultures. There was a pronounced elevation of immunoreactive ACTH content in both cells and media and an enhanced secretory response to stimulation of cultures with stalk-median eminence extract containing cortiocotropin releasing (CRF) activity. Some cells identified as corticotrophs by a specific immunostaining incorporated tritiated thymidine into their nuclei suggesting their ability to enter the cell cycle. The relatively smaller increase in number of ACTH cells and the considerably higher ACTH producing capacity of the corticotrophs after adrenalectomy seem to be inconsistent with the quantal response model of hormone secretion recently introduced by Rodbard.

A serine-proteinase inhibitor (Boc-D-Phe-Pro-Arg-H) inhibits the secretion of adrenocorticotropin- and β-endorphin-immunoreactive peptides in vitro

The effects of a potent serine protease inhibitor, Boc-D-Phe-Pro-Arg-H, on the secretion and content of ACTH and beta-endorphin-like immunoreactivities of cultured rat anterior pituitary cells were studied. Basal release of the hormones was not affected by the drug but secretion in response to stalk-median eminence extracts was inhibited. The inhibitor did not effect the amount of ACTH or beta-endorphin in the medium plus the cells, nor did it change the ratio of the lipotropin and beta-endorphin-like immunoreactivities. Thus the compound seems to affect the release of ACTH and beta-endorphin, but not the processing of lipotropin to beta-endorphin.

Various proteinase inhibitors decrease prolactin and growth hormone release by anterior pituitary cells

Proteinase inhibitors were tested for their ability to inhibit prolactin (PRL) and growth hormone (GH) release by cultured anterior pituitary cells of the rat. Inhibitors of microbial origin (chymostatin, elastatinal, leupeptin) had either no or a moderate effect on hormone release while some tripeptide aldehydes, especially those with lysine at their C terminus, inhibited markedly PRL and to a lesser extent GH release. Boc-DPhe-Phe-lysinal was the most effective on lactotrophs inhibiting PRL release more than 50% at 10(-4) M. The site(s) of action of tripeptide aldehydes remain to be elucidated.

Major metabolic pathways and hormone production in unstimulated monolayer cultures of the rat anterior pituitary

SummaryIn cell cultures derived from anterior pituitary glands of rats, enzyme activities of cell homogenates and hormone (GH, PRL, LH, and FSH) content of the culture media were measured. Sex differences in enzyme activities representing major metabolic pathways (citrate cycles, pentose cycles, and glycolysis) were demonstrated both in freshly dispersed cells and in 8-day-old cultures; in cultures of both sexes enzyme activities increased during cultivation.In cultures derived from female rats, cell protein doubled by the 12th day and remained constant for up to the 24th day in culture, whereas enzyme activities showed changes suggesting that cell metabolism shifted to anaerobic glycolysis during cultivation. In the culture media the presence of four pituitary hormones was demonstrated for as long as 3 weeks of cultivation with variable secretion dynamics; the release of gonadotropic hormones diminished gradually whereas that of GH remained constant and PRL levels increased with time. These results indicate that under strictly defined culture conditions pituitary cells may function in spite of profound metabolic changes.

Corticotroph, somatotroph and mammotroph cell kinetics in the postnatal infant female rat

The aim of this work was to detect if hypothalamic-pituitary maturation was accompanied by significant proliferation changes in differentiated pituitary cell pools. For this purpose, pituitary corticotroph (Ct), mammotroph (Mt) and somatotroph (St) proliferation activities were scanned in intact female rats during the postnatal (P) period (1–35 postnatal days). The techniques of tritiated thymidine labelling, immunostaining and autoradiography were combined to visualize DNA synthesis of hormone containing cells. Immunoreactive cell densities were measured using image analysis, and double labelled cells were counted. Corticotroph proliferation activity increased significantly on day P12, followed by an increase in the Ct proportion on days P13–14. This is the first observation of a spontaneous change of corticotroph proliferation at the end of the stress nonresponsive period. The mammotroph density and proliferation rate increased gradually during postnatal maturation, until the Mt pool overran other cell types of the female hypophysis on day 35. The somatotroph pool was the most numerous until day P20; the proliferation rate remained constant while St proportions increased reaching a plateau between days P13 and 20, then decreased to the adult level. Each cell type examined showed a characteristic, individual density and proliferation pattern.

The tripeptide aldehyde, Boc-DPhe-Phe-Lysinal, is a novel Ca2+ channel inhibitor in pituitary cells

The effect of Boc-DPhe-Phe-Lysinal (Boc-DPPL) on the 45Ca2+ uptake of rat anterior pituitary monolayer cultures was investigated. The compound decreased the basal Ca2+ uptake at 3 x 10(-4) mol/l. The 45Ca2+ uptake stimulated by potassium-induced depolarization was more sensitive to Boc-DPPL inhibition, a slight decrease was seen with 3 x 10(-6) mol/l and there was a half maximal inhibition at 3 x 10(-5) mol/l. Boc-DPPL is known to inhibit pituitary hormone release in similar concentrations, an effect might also be due to its calcium antagonist property.

An intracellular factor (DCI) controlling differentiation and cell division

Abstract Previous concepts correlating cell differentiation and division are elaborated, and some experimental data are presented which strongly suggest the existence of an inhibitor of the mitotic cycle that is produced by cells during the course of differentiation.

DNA synthesis in lactotrophs of primary rat anterior pituitary cell cultures. Effects of thyroliberin, bromocriptine and somatostatin.

SummaryProlactin immunostaining in combination with thymidine autoradiography was used to characterize changes in the DNA-synthesizing activity of lactotrophs in primary monolayer cultures of the rat anterior pituitary gland treated for 3 days with thyroliberin (TRH), somatostatin (SRIF) and bromocriptine (CB 154). The number of lactotrophs labelled with3H-thymidine within the total pool of labeled pituitary cells was used to estimate DNA synthesis in prolactin-producing cells. TRH (10 ng/ml) stimulated DNA synthesis in the whole population of cultured cells but not in lactotrophs. TRH only weakly counteracted the noticeable inhibitory effect of CB 154 (0.75 μM/l) on thymidine incorporation into lactotrophs. SRIF (20 ng/ml) inhibited DNA synthesis in lactotrophs to a lesser extent than CB 154. The combination of methods used in this paper may be useful for studying the selective effects of regulators on the proliferative activity of various pituitary cell types in vivo and in culture.