PMID- 11356667
OWN - NLM
STAT- MEDLINE
DCOM- 20010621
LR  - 20131121
IS  - 0013-7227 (Print)
IS  - 0013-7227 (Linking)
VI  - 142
IP  - 6
DP  - 2001 Jun
TI  - Gonadotropins decrease estrogen receptor-beta messenger ribonucleic acid 
      stability in rat granulosa cells.
PG  - 2230-7
AB  - We have previously shown that the preovulatory LH surge down-regulates estrogen 
      receptor-beta (ERbeta) messenger RNA (mRNA) levels selectively in the granulosa 
      cells of preovulatory follicles. To gain insight into the underlying mechanisms, 
      we examined whether the LH-induced loss of ERbeta mRNA expression in rat 
      granulosa cells is attributable to the hormone-induced changes at the level of 
      transcription and/or mRNA degradation. When the rate of ERbeta gene transcription 
      was assessed in cultured granulosa cells, by nuclear run-off assays, we observed 
      only a marginal effect of hCG on ERbeta gene transcription. In contrast, when 
      ERbeta mRNA levels were estimated in granulosa cells that were cultured in the 
      presence of 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB), an RNA 
      synthesis inhibitor, we observed a significant inhibitory effect of human CG 
      (hCG) on ERbeta mRNA expression at a magnitude similar to that observed in the 
      absence of DRB. Forskolin (FSK) and 2-O-tetradecanol-phorbol-13-acetate (TPA), 
      pharmacological agents that mimic LH actions in granulosa cells, also showed 
      similar effects. Thus, these results suggest that LH decreases ERbeta mRNA 
      expression in the granulosa cells of preovulatory follicles, primarily by 
      destabilizing the preexisting ERbeta mRNA. We next determined the decay rate of 
      the ERbeta mRNA in granulosa cells that were cultured in the presence of DRB and 
      additional hCG, FSK, or TPA for various time periods, by estimating ERbeta mRNA 
      levels, using semiquantitative RT-PCR assays and subsequent linear regression 
      analyses. The half-life of the ERbeta mRNA in the presence of vehicle was 17.87 
      +/- 1.2 h (n = 4). hCG dramatically decreased the half-life of the ERbeta mRNA 
      (4.85 +/- 0.49 h, n = 4). Similarly, both FSK and TPA decreased the half-life of 
      the ERbeta mRNA to 3.57 +/- 0.31 h and 4.02 +/- 0.13 h, respectively. We extended 
      these findings by examining whether the LH-induced down-regulation of the ERbeta 
      mRNA is cycloheximide-sensitive. When granulosa cells were cultured in the 
      presence of cycloheximide, a protein synthesis inhibitor, the inhibitory effects 
      of hCG, FSK, and TPA on ERbeta mRNA levels were abolished. Similar results were 
      obtained in the presence or absence of DRB, indicating that the hormone-induced 
      destabilization of the ERbeta mRNA is coupled with translation processes. Taken 
      together, our results demonstrate that LH decreases ERbeta mRNA expression, 
      predominantly at the posttranscriptional level, in a cycloheximide-sensitive 
      manner.
FAU - Guo, C
AU  - Guo C
AD  - Department of Physiology, University of Kentucky, Lexington 40536-0084, USA.
FAU - Savage, L
AU  - Savage L
FAU - Sarge, K D
AU  - Sarge KD
FAU - Park-Sarge, O K
AU  - Park-Sarge OK
LA  - eng
GR  - HD-01135/HD/NICHD NIH HHS/United States
GR  - HD-30719/HD/NICHD NIH HHS/United States
GR  - HD-36879/HD/NICHD NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Endocrinology
JT  - Endocrinology
JID - 0375040
RN  - 0 (Chorionic Gonadotropin)
RN  - 0 (Estrogen Receptor beta)
RN  - 0 (Nucleic Acid Synthesis Inhibitors)
RN  - 0 (RNA, Messenger)
RN  - 0 (Receptors, Estrogen)
RN  - 1F7A44V6OU (Colforsin)
RN  - 53-85-0 (Dichlororibofuranosylbenzimidazole)
RN  - 9002-67-9 (Luteinizing Hormone)
RN  - NI40JAQ945 (Tetradecanoylphorbol Acetate)
SB  - IM
MH  - Animals
MH  - Cells, Cultured
MH  - Chorionic Gonadotropin/pharmacology
MH  - Colforsin/pharmacology
MH  - Dichlororibofuranosylbenzimidazole/pharmacology
MH  - Drug Stability
MH  - Estrogen Receptor beta
MH  - Female
MH  - Granulosa Cells/*chemistry
MH  - Luteinizing Hormone/*pharmacology
MH  - Nucleic Acid Synthesis Inhibitors/pharmacology
MH  - RNA, Messenger/*analysis
MH  - Rats
MH  - Rats, Sprague-Dawley
MH  - Receptors, Estrogen/*genetics
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - Tetradecanoylphorbol Acetate/pharmacology
EDAT- 2001/05/18 10:00
MHDA- 2001/06/22 10:01
CRDT- 2001/05/18 10:00
PHST- 2001/05/18 10:00 [pubmed]
PHST- 2001/06/22 10:01 [medline]
PHST- 2001/05/18 10:00 [entrez]
AID - 10.1210/endo.142.6.8102 [doi]
PST - ppublish
SO  - Endocrinology. 2001 Jun;142(6):2230-7. doi: 10.1210/endo.142.6.8102.