title: Heat stress diminishes gonadotropin receptor expression and enhances susceptibility to apoptosis of rat granulosa cells.
abstract: Heat stress inhibits ovarian follicular development in mammalian species. We hypothesized that heat stress inhibits the function of follicular granulosa cells and suppresses follicular development. To test this, immature female rats were injected with pregnant mare serum gonadotropin (PMSG) at 48 h after the start of temperature treatment (control: 25 degrees C, 50% RH; heat stress: 35 degrees C, 70% Relative Humidity). The ovaries and granulosa cells of follicles at different developmental stages were analyzed for gonadotropin receptor levels and aromatase activity; estradiol levels were measured in follicular fluid. Before injection, heat stress diminished only the amount of FSH receptor on granulosa cells of antral follicles. During PMSG-stimulated follicular development, heat stress strongly inhibited gonadotropin receptor levels and aromatase activity in granulosa cells, and estradiol levels in the follicular fluid of early antral, antral and preovulatory follicles. To examine apoptosis and mRNA levels of bcl-2 and bax in granulosa cells, follicles harvested 48 h after PMSG injection were cultured in serum-free conditions. Heat-stressed granulosa cells showed a time-dependent increase in apoptosis. The bcl-2 mRNA levels were similar in control and heat-stressed granulosa cells; bax mRNA levels were increased in heat-stressed granulosa cells. According to these results, heat stress inhibits expression of gonadotropin receptors in granulosa cells and attenuates estrogenic activity of growing follicles, granulosa cells of heat-stressed follicles are susceptible to apoptosis, and the bcl2/bax system is not associated with heat-stress-induced apoptosis of granulosa cells. Our study suggests that decreased numbers and function of granulosa cells may cause ovarian dysfunction in domestic animals in summer.
authors.affiliation: Institute of Agriculture and Forestry, University of Tsukuba, Tsukuba, Ibaraki 305-8572, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Shimizu
authors.forename: Izumi
authors.initials: I
authors.lastname: Ohshima
authors.forename: Manabu
authors.initials: M
authors.lastname: Ozawa
authors.forename: Satoko
authors.initials: S
authors.lastname: Takahashi
authors.forename: Atsushi
authors.initials: A
authors.lastname: Tajima
authors.forename: Masayuki
authors.initials: M
authors.lastname: Shiota
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Miyazaki
authors.forename: Yukio
authors.initials: Y
authors.lastname: Kanai
mesh_terms: D000818:Animals; D017209:Apoptosis; D018929:Cell Culture Techniques; D004958:Estradiol / Q000032:analysis; D005260:Female; D015571:Follicular Fluid / Q000737:chemistry; D019254:Genes, bcl-2; D006064:Gonadotropins, Equine / Q000494:pharmacology; D006107:Granulosa Cells / Q000166:cytology / Q000378:metabolism*; D006358:Hot Temperature / Q000009:adverse effects*; D011485:Protein Binding; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017208:Rats, Wistar; D011962:Receptors, FSH / Q000378:metabolism; D011967:Receptors, Gonadotropin / Q000378:metabolism*; D011974:Receptors, LH / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D051028:bcl-2-Associated X Protein
chemical_list: C494828:Bax protein, rat; D006064:Gonadotropins, Equine; D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger; D011962:Receptors, FSH; D011967:Receptors, Gonadotropin; D011974:Receptors, LH; D051028:bcl-2-Associated X Protein; D004958:Estradiol