Nuclear acceptor sites for estrogen-receptor complexes in the liver of the turtle, Chrysemys picta. I. Sexual differences, species specificity and hormonal dependency.
Hepatic estrogen receptors (ERs) of the female turtle, Chrysemys picta, when complexed with [3H]estradiol ([3H]E2), were shown to bind specifically to liver chromatin isolated from the same species. The binding of the [3H]E2 receptor complex to chromatin requires both the steroid ligand and the receptor protein. Maximal binding occurred within 60-70 min of incubation at 4 degrees C in a Tris buffer containing 0.1 M KCl. The binding of the [3H]E2 receptor complex to intact chromatin was saturable, whereas the binding to turtle or calf thymus DNA remained linear. Scatchard analyses revealed more estrogen receptor binding sites on hepatic chromatin isolated from female turtles than that prepared from the males (binding capacities: female chromatin = 67.9 +/- 6.8 fmol/mg DNA equivalent; male chromatin = 28.5 +/- 2.5 fmol/mg DNA equivalent). Furthermore, the [3H]E2 receptor complex was bound with a higher affinity to female chromatin than to male chromatin (association constants: female chromatin = 11.7 +/- 2.7 X 10(10) M-1; male chromatin = 2.5 +/- 0.7 X 10(10) M-1). In contrast to turtle hepatic [3H]E2 receptors, ERs in rat liver or mouse uterine cytosol exhibited little binding affinity for hepatic chromatin isolated from the turtle. Tissue specificity was demonstrated in the interaction of the [3H]E2 receptor complex and chromatin; high affinity, saturable binding of the [3H]E2 receptor complex was only observed on chromatin isolated from the liver but not on those prepared from the heart, kidney and muscle. A 3- to 4-fold increase in the number of hepatic chromatin [3H]E2 receptor binding sites was observed in 21-day ovariectomized or hypophysectomized female (capacities = 209.3 +/- 6.1 and 270 +/- 10.1 fmol/mg DNA equivalent, respectively). It is postulated that [3H]E2 receptor binding sites on the chromatin of intact females are partially 'masked', and removal of a gonadal and/or pituitary factor(s) unveils additional binding sites on the female chromatin. This paper is first to report the presence of high affinity, species- and tissue-specific acceptor sites on the liver chromatin of a reptilian species. The fact that the levels and properties of these acceptor sites are dependent on the sex and hormonal state of the animal suggests that they may play a role in the regulation of hepatic estrogen responsiveness and vitellogenesis in this species.
Yu
M S
MS
Department of Biology, Tufts University, Medford, MA 02155.
Ho
S M
SM
D000818:Animals; D001665:Binding Sites / Q000187:drug effects; D002417:Cattle; D002467:Cell Nucleus / Q000378:metabolism*; D002843:Chromatin / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D004958:Estradiol / Q000494:pharmacology*; D006801:Humans; D007668:Kidney / Q000378:metabolism; D008099:Liver / Q000378:metabolism*; D008297:Male; D009132:Muscles / Q000378:metabolism; D009206:Myocardium / Q000378:metabolism; D051381:Rats; D011960:Receptors, Estrogen / Q000378:metabolism*; D012737:Sex Factors; D013045:Species Specificity; D013950:Thymus Gland / Q000378:metabolism; D014426:Turtles
D002843:Chromatin; D011960:Receptors, Estrogen; D004958:Estradiol; D004247:DNA