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a Department of On/Gyn, Johns Hopkins University School of Medicine, Baltimore, Maryland 21201
b Division of Reproductive Biology, Department of Population Dynamics, Johns Hopkins University, Baltimore, Maryland 21201
c Vincent Center for Reproductive Biology, Department of Ob/Gyn, Massachusetts General Hospital/ Harvard Medical School, Boston, Massachusetts 02114
d Department of Anatomy and Human Biology, The University of Western Australia, Nedlands, Perth, Western Australia, Australia
In the rabbit, estradiol is the primary luteotropic hormone. Estradiol withdrawal results in a rapid decline in serum progesterone and eventually in corpus luteum (CL) regression. The objective of this study was to determine whether estradiol modulates luteal cell apoptosis. In the first experiment, rabbits were randomly assigned to one of five experimental groups. An empty capsule (control) or estradiol-filled Silastic capsule was inserted s.c. on Day 0 of pseudopregnancy (day of hCG administration). On Day 11 of pseudopregnancy, some of the group I (control) and group II (estradiol capsule) rabbits were subjected to laparotomy, and one ovary from each rabbit was perfused in vitro to determine progesterone secretion rates. The CL from the contralateral ovary were dissected, snap-frozen, and stored at -70°C until analyzed for internucleosomal DNA cleavage (apoptosis). Estradiol-containing capsules were removed from some of the remaining rabbits on Days 8, 9, and 10 to initiate estradiol deprivation. Rabbits were then subjected to laparotomy 24, 48, or 72 h after capsule removal (groups III, IV, and V, respectively), and ovaries or CL were processed as described above. Deprivation of estradiol for 24 (group III), 48 (group IV), or 72 (group V) h in vivo reduced in vitro progesterone secretion rates by more than 90% as compared to that in ovaries collected from estradiol capsule-intact animals. After in vivo endogenous estradiol suppression, withdrawal of exogenous estradiol resulted in luteal cell apoptosis, which increased in a time-dependent manner. Northern blot analysis revealed an increase in bax mRNA levels and a decrease in bcl-x mRNA levels coincident with luteal cell apoptosis induced by estradiol withdrawal. These data demonstrate that changes in progesterone production caused by estradiol exposure and deprivation are in part related to luteal cell apoptosis, and alterations in the expression of bcl-2 gene family members may be one of the mechanisms by which estradiol exerts its luteotropic effect in the rabbit CL.
2 Correspondence. FAX: 61–8-9380–1051; dharma{at}anhb.uwa.edu.au
3 Current address: 2804 Fountain Grove Terrace, Olney, MD 20832.
4 Current address: Department of Ob/Gyn, The University of Tokyo Faculty of Medicine, 7–3-1 Hongo, Bunkyo-ku, Tokyo 113, Japan.
5 Current address: 2001 Marcus Avenue, Suite N213, Lake Success, NY 11042.
6 Current address: Department of Gyn/Ob, Ramathibodi Hospital, Mahidol University School of Medicine, Bangkok, Thailand.
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