Reptilian (Chrysemys picta) hepatic progesterone receptors: relationship to plasma steroids and the vitellogenic cycle.
In non-mammals, estrogen-induced yolk precursors produced by the adult female liver are the main nutritional source for development. Evidence exists that progesterone exerts counter-regulatory effects on estrogen-induced vitellogenesis, and we have used the turtle model (Chrysemys picta) to study changes in hepatic progesterone receptor during the vitellogenic cycle. Using radioligand methods, we show that high and lower affinity binding sites are present in the cytosolic but not nuclear extracts. The lower affinity site is detectable at all times of the year; the high affinity site is mainly observed during non-vitellogenic periods and does not correlate with plasma estrogen. DNA-cellulose chromatography shows that PR-A is present in spring, summer, and winter, and that PR-B is down-regulated except in animals which recently laid eggs. Western blots confirm the presence of PR in all months, but PR-A (88 kDa) is the dominant isoform. PR-B (125 kDa) is well correlated with the luteal phase, winter and fall. Immunocytochemical studies show that PR is nuclear in location, and nuclear heat shock protein 90 (hsp90) is present. Competitive binding studies of both sites reveal that progesterone is the most effective ligand for both, followed by pregnenolone, deoxycorticosterone, and R5020. RU 486 does not bind to the high affinity site but binds moderately well to the lower affinity site. This study suggests that progesterone receptor isoforms are differentially expressed and may be involved as transcriptional regulators of hepatic function outside the periods of active vitellogenesis in the turtle.
Giannoukos
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Department of Biology, Boston University, MA 02215, USA.
Callard
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D000818:Animals; D001667:Binding, Competitive; D002457:Cell Extracts; D002467:Cell Nucleus / Q000737:chemistry / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D003900:Desoxycorticosterone / Q000378:metabolism; D004958:Estradiol / Q000097:blood; D005260:Female; D008099:Liver / Q000502:physiology*; D015735:Mifepristone / Q000378:metabolism; D011284:Pregnenolone / Q000378:metabolism; D011374:Progesterone / Q000097:blood* / Q000378:metabolism; D045167:Progesterone Congeners / Q000378:metabolism; D011397:Promegestone / Q000378:metabolism; D011980:Receptors, Progesterone / Q000032:analysis / Q000378:metabolism*; D012621:Seasons; D014426:Turtles; D014818:Vitellogenesis / Q000502:physiology*
D002457:Cell Extracts; D045167:Progesterone Congeners; D011980:Receptors, Progesterone; D015735:Mifepristone; D003900:Desoxycorticosterone; D011374:Progesterone; D004958:Estradiol; D011284:Pregnenolone; D011397:Promegestone