REGULATION OF PROTOONCOGENE EXPRESSION AND DEOXYRIBONUCLEIC-ACID SYNTHESIS IN GRANULOSA-CELLS OF PERIFUSED IMMATURE RAT OVARIES

被引:27
作者
DELIDOW, BC
LYNCH, JP
WHITE, BA
PELUSO, JJ
机构
[1] UNIV CONNECTICUT,CTR HLTH,DEPT OBSTET & GYNECOL,FARMINGTON,CT 06032
[2] UNIV CONNECTICUT,CTR HLTH,DEPT ANAT,FARMINGTON,CT 06032
关键词
D O I
10.1095/biolreprod47.3.428
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
The present series of experiments examined the effects of follicle-stimulating hormone (FSH) and insulin (IN) on granulosa cell (GC) proto-oncogene expression and DNA synthesis. In the first study, GCs were harvested from immature rat ovaries after 15, 30, or 60 min of perifusion and DNA synthesis (H-3-thymidine incorporation) and proto-oncogene mRNA levels were determined. The presence of c-myc and c-fos proteins was localized within GCs immunocytochemically. GCs of control ovaries exhibited modest levels of DNA synthesis and proto-oncogene expression. FSH/IN not only stimulated DNA synthesis but also increased c-myc, c-fos, and c-jun mRNA levels and the percentage of cells staining for c-fos and c-myc proteins. The protein kinase inhibitor, 2-aminopurine (2-AP), inhibited the FSH/IN-induced increases in c-myc and c-fos mRNA levels, the percentage of cells staining for Myc and Fos protein, and DNA and protein synthesis. The effects of 48 h of perifusion with FSH in the presence or absence of IN were also examined. These treatments were selected because after 48 h of continuous exposure to FSH alone, estradiol-17-beta (E2) secretion is enhanced and H-3-thymidine incorporation is inhibited. Conversely, FSH/IN maintains H-3-thymidine incorporation for up to 48 h of perifusion culture without stimulating E2 (Peluso et al., Endocrinology 1991; 128:191-196). After 48 h of perifusion, both FSH and FSH/IN stimulated c-fos mRNA and protein levels. However, high levels of c-jun mRNA and protein were detected only within GCs of FSH/IN-treated ovaries. These observations support the concept that c-myc, c-fos; and c-jun are involved in regulating GC proliferation and that c-jun is selectively expressed in mitotically active GCs.
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页码:428 / 435
页数:8
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