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Submitted on February 28, 2003
Accepted on July 25, 2003
1 Laboratoire de Biochimie, UPRES EA 2608, USC INRA, CHU Côte de Nacre, 14032 Caen cedex, France.
* To whom correspondence should be addressed. E-mail: benhaim-a{at}chu-caen.fr.
In rabbit granulosa cells, two cytochrome P450 aromatase (P450 arom) mRNAs issued from promoter II were described: a full-length and a truncated transcript. Western blot analysis showed two P450 arom proteins with apparent molecular weights of 53 kDa and 46 kDa, which are consistent with the predicted theoretical sizes of proteins encoded by these two transcripts. To examine the involvement of the truncated transcript in the regulation of P450 arom gene expression, the level of each transcript was specifically quantified in cultured granulosa cells by competitive quantitative RT-PCR. FSH induced a dose-dependent increase in both estradiol production and P450 arom mRNAs levels with a much more enhancement in the full-length mRNA. The half-life of the transcripts could not explain this differential regulation. Upon dibutytryl cyclic AMP (db cAMP) stimulation, the full-length mRNA was less abundant than the truncated one. In contrast, Western blot analysis revealed a stimulation of the 53 kDa protein content whereas the 46 kDa protein amount was apparently unaffected. TGF
in FSH-stimulated conditions decreased both estradiol production and P450 arom transcripts levels. TGF
did not modify estradiol production and aromatase protein amounts induced by db cAMP whereas the two P450 arom mRNAs levels were increased. In conclusion, we report for the first time that a protein encoded by a truncated P450 arom mRNA could be involved in the regulation of estrogen production. Moreover, we show that the two P450 arom mRNAs are regulated in a differential manner probably through hormonal control of the alternative splicing.
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