Decreased progesterone levels and progesterone receptor antagonists promote apoptotic cell death in bovine luteal cells

Bo R. Rueda, Isabel R. Hendry, William J. Hendry, Fredrick Stormshak, O. D. Slayden, John S. Davis

Research output: Contribution to journalArticle

100 Citations (Scopus)

Abstract

We tested the hypothesis that progesterone (P4) acts at a local level to inhibit luteal apoptosis. Initial experiments employed aminoglutethimide, a P450 cholesterol side-chain cleavage inhibitor, to inhibit steroid synthesis. Cultured bovine luteal cells were treated with aminoglutethimide (0.15 mM) ± P4 (500 ng/ml) for 48 h. Luteal cells were recovered and snap frozen for isolation and analysis of oligonucleosomal DNA fragmentation or fixed for morphological analysis. Medium was collected for analysis of P4 levels by RIA. Aminoglutethimide inhibited P4 synthesis by > 95% and increased the level of apoptosis as evidenced by 32P-labeled oligonucleosomal DNA fragmentation (> 40%). P4 supplementation inhibited the onset of apoptosis that was induced by aminoglutethimide. These data were further supported by morphological assessment of apoptotic cells utilizing a Hoechst staining technique and together strongly suggest that P4 has anti- apoptotic capacity. Using reverse transcription-polymerase chain reaction, we were able to isolate a 380-base pair cDNA from the bovine corpus luteum (CL) that was 100% homologous to the progesterone receptor (PR) previously found in bovine oviductal tissue. Furthermore, PR transcripts were present in large and small luteal cells. Immunohistochemistry also revealed that PR protein was present in both large and small luteal cells. To determine whether the anti-apoptotic effect of P4 was regulated at the receptor level, luteal cells were cultured in the presence of PR antagonists, RU-486 and onapristone, for 48 h. Both antagonists caused approximately a 40% increase in 32P. labeled oligonucleosomal DNA fragmentation. Interestingly, there was no difference (P ≥ 0.05) in P4 levels after treatment with PR antagonists. These observations support the concept that P4 represses the onset of apoptosis in the CL by a PR-dependent mechanism.

Original languageEnglish (US)
Pages (from-to)269-276
Number of pages8
JournalBiology of reproduction
Volume62
Issue number2
DOIs
StatePublished - Jan 1 2000

Fingerprint

Luteal Cells
Progesterone Receptors
Progesterone
Aminoglutethimide
Cell Death
Corpus Luteum
DNA Fragmentation
Apoptosis
Mifepristone
Base Pairing
Reverse Transcription
Complementary DNA
Immunohistochemistry
Steroids
Cholesterol
Staining and Labeling
Polymerase Chain Reaction

ASJC Scopus subject areas

  • Reproductive Medicine
  • Cell Biology

Cite this

Decreased progesterone levels and progesterone receptor antagonists promote apoptotic cell death in bovine luteal cells. / Rueda, Bo R.; Hendry, Isabel R.; Hendry, William J.; Stormshak, Fredrick; Slayden, O. D.; Davis, John S.

In: Biology of reproduction, Vol. 62, No. 2, 01.01.2000, p. 269-276.

Research output: Contribution to journalArticle

Rueda, Bo R. ; Hendry, Isabel R. ; Hendry, William J. ; Stormshak, Fredrick ; Slayden, O. D. ; Davis, John S. / Decreased progesterone levels and progesterone receptor antagonists promote apoptotic cell death in bovine luteal cells. In: Biology of reproduction. 2000 ; Vol. 62, No. 2. pp. 269-276.
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