Growth hormone-releasing hormone and glucocorticoids determine the balance between luteinising hormone (LH) beta- and LH beta/follicle-stimulating hormone beta-positive gonadotrophs and somatotrophs in the 14-day-old rat pituitary tissue in aggregate cell culture.
Fourteen-day-old rat pituitary tissue represents an attractive model for studying cell population dynamics, particularly of gonadotrophs. Prolonged three-dimensional culture in serum- and hormone-free medium causes a striking decline in somatotroph abundance but a several-fold rise in monohormonal LH beta-positive cell number, whereas bihormonal gonadotrophs almost disappear. In the present study, we investigated whether these changes are inter-related by examining the effects of growth hormone-releasing hormone (GHRH) and glucocorticoids, two protagonist regulators of somatotrophs. Cells were identified by single cell reverse transcriptase-polymerase chain reaction (RT-PCR) and immunofluorescence. Supplementation of the cultures for 2 weeks with GHRH (1 nm) did not augment the proportion of somatotrophs, but expanded the nonhormonal cell population. GHRH reduced the proportion of monohormonal luteinising hormone (LH)beta mRNA positive cells to approximately 50% of control, although the effect was not seen when these cells were visualised by immunostaining. Supplementation of the cultures with dexamethasone (4 nM) for 3 weeks partially rescued LH beta/follicle-stimulating hormone beta cells and fully rescued the GH mRNA cells in parallel with a decline in nonhormonal cell abundance, but strongly reduced bromodeoxyuridine labelling of GH-immunoreactive cells. As studied by patch-clamp single cell RT-PCR at the start of culture, GHRH caused an acute rise in intracellular [Ca(2+)] in some monohormonal GH cells, but at a higher incidence in cells expressing LH beta mRNA, alone or in combination with GH mRNA and/or pro-opiomelanocortin (POMC) mRNA. The present data suggest that, in the 14-day-old rat pituitary, the majority of GHRH target cells are cells expressing LH beta mRNA alone or in combination with GH and/or POMC mRNA. The data show co-regulation of gonadotroph and somatotroph population sizes by glucocorticoids and GHRH, with the former preserving bihormonal gonadotrophs and the latter repressing LH beta-only cell abundance. GHRH may not expand the somatotroph population unless glucocorticoid hormone is present to maintain terminal differentiation.
Pals
K
K
Laboratory of Cell Pharmacology, University of Leuven, Medical School, Campus Gasthuisberg (O&N), Leuven, Belgium.
Roudbaraki
M
M
Denef
C
C
D000367:Age Factors; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002449:Cell Aggregation; D002452:Cell Count; D002454:Cell Differentiation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003907:Dexamethasone / Q000494:pharmacology; D005260:Female; D037201:Follicle Stimulating Hormone, beta Subunit / Q000235:genetics / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000187:drug effects; D005938:Glucocorticoids / Q000494:pharmacology*; D052681:Gonadotrophs / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D013006:Growth Hormone / Q000235:genetics / Q000378:metabolism; D013007:Growth Hormone-Releasing Hormone / Q000494:pharmacology*; D037101:Luteinizing Hormone, beta Subunit / Q000235:genetics / Q000378:metabolism*; D010902:Pituitary Gland / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D011333:Pro-Opiomelanocortin / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D052683:Somatotrophs / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism
D037201:Follicle Stimulating Hormone, beta Subunit; D005938:Glucocorticoids; D037101:Luteinizing Hormone, beta Subunit; D012333:RNA, Messenger; D011333:Pro-Opiomelanocortin; D003907:Dexamethasone; D013006:Growth Hormone; D013007:Growth Hormone-Releasing Hormone; D002118:Calcium