Estradiol regulation of follistatin and inhibin alpha- and beta(B)-subunit mRNA in avian granulosa cells.
Estradiol modulation of granulosa cell growth and regulation of follistatin and inhibin alpha- and beta(B)-subunit mRNA were investigated in cultured chicken granulosa cells. Granulosa cells were isolated and pooled according to size from the F(4) + F(5), small yellow (SYF), and large white (LWF) follicles. Isolated and dispersed granulosa cells were then cultured in the absence or presence of 1 x 10(-5) M 17 beta-estradiol. In Experiment 1 (n = 4 replications) the effect of estradiol on the growth of granulosa cells from the different-sized follicles was examined at 24 and 48 h of culture. Untreated and treated granulosa cells from all three follicle sizes proliferated during culture, and cell viability for all cultures was over 95% throughout the experiment. After 48 h the untreated cultures for all follicle types had 1.6 to 2.2 times (P < 0.05) more cells than the estradiol-treated cultures. In Experiment 2 (n = 3 replications), the cultures were terminated at 4 and 24 h after plating. Follistatin mRNA levels were higher in estradiol-treated cells at 24 h in F(4) + F(5) follicles, at 4 and 24 h in the SYF, and at 4 h in the LWF. beta(B)-subunit mRNA levels were also increased by estradiol at 4 h in F(4) + F(5) cells and at 4 and 24 h in the LWF. Steady state mRNA levels for the alpha-subunit were higher (P < 0.05) in estradiol-treated cultures at 4 and 24 h in F(4) + F(5) follicles and at 24 h in the SYF. Immunoreactive alpha-subunit protein, however, was not increased by estradiol treatment. Thus, whereas estradiol inhibited granulosa cell growth, it exerted a generally stimulatory effect on the expression of FS and the inhibin alpha- and beta(B)-subunit mRNA.
Davis
A J
AJ
Department of Animal Science, Cornell University, Ithaca, New York 14853, USA.
Brooks
C F
CF
Johnson
P A
PA
D000818:Animals; D002452:Cell Count; D002478:Cells, Cultured; D002645:Chickens / Q000378:metabolism*; D004958:Estradiol / Q000494:pharmacology*; D005260:Female; D038681:Follistatin; D005786:Gene Expression Regulation / Q000187:drug effects*; D006023:Glycoproteins / Q000235:genetics*; D006107:Granulosa Cells / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D007265:Inhibins / Q000235:genetics*; D011374:Progesterone / Q000096:biosynthesis; D012333:RNA, Messenger / Q000032:analysis
D038681:Follistatin; D006023:Glycoproteins; D012333:RNA, Messenger; D011374:Progesterone; D004958:Estradiol; D007265:Inhibins