title: Ovarian steroids modulate responsiveness to dopamine and expression of G-proteins in lactotropes.
abstract: The effects of chronic ovarian steroid treatment on the secretory activity of individual lactotropes and the mechanisms modulating their responsiveness to dopamine (DA) were studied. Female rats were ovariectomized (OVX) and implanted with Silastic capsules containing progesterone (P4), 17beta), 17beta-estradiol (E2) or both E2 (E2+P4). Ten days after surgery, anterior pituitaries were enzymatically dispersed and the reverse hemolytic plaque assay (RHPA) was performed to assess the release of prolactin (PRL) from individual lactotropes. RHPA was combined with immunocytochemistry (ICC) for PRL, Galphas or Gialpha3/Galphao proteins. E2 treatment alone or in combination with P4 increased the percentage of immunoreactive lactotropes among anterior pituitary cells. Incidence of active (plaque-forming) lactotropes, however, was increased both in P4-, and E2-treated rats and E2+P4 treatment increased it even further. While P4 treatment did not affect the frequency distribution of lactotropes, both E2 and E2+P4 treatments increased the large plaque-forming lactotrope population. This increase was reflected by the significantly greater mean plaque areas of lactotropes from E2- and E2+P4-treated rats compared to OVX or P4-treated animals. The responsiveness of lactotropes to DA from P4-treated rats did not differ from that of OVX rats: thus challenge with 1 microM DA inhibited the release of PRL, while 100 pM DA had no effect. E2 and E2+P4 treatments, however, profoundly changed the lactotrope's responsiveness: challenge with 1 microM DA had no effect and 100 pM DA resulted in moderate stimulation of PRL release in E2+P4 rats. Double-label ICC revealed that ovarian steroid treatments did not affect the expression of Galphas in lactotropes. The incidence of Gialpha3/Galphao-immunoreactive lactotropes, however, decreased after E2 treatment, alone or in combination with P4. Although expression of Galphas was similar in all plaque-forming cells regardless of plaque size, lactotropes expressing Gialpha3/Galphao were more likely to form small plaques in all treatment groups. These data suggest: (1) ovarian steroid treatment recruits quiescent lactotropes to release PRL; (2) E2 treatment alone or in combination with P4 increases the amount of PRL rleased by individual lactotropes; (3) E2-induced alterations in the frequency distribution and lactotrope responsiveness to DA may be due in part to a decreased expression of Gialpha3/Galphao.
authors.affiliation: Department of Biological Science, Program in Neuroscience, The Florida State University, Tallahassee, Fla., USA.
authors.forename: J D
authors.initials: JD
authors.lastname: Livingstone
authors.forename: A
authors.initials: A
authors.lastname: Lerant
authors.forename: M E
authors.initials: ME
authors.lastname: Freeman
mesh_terms: D000818:Animals; D002452:Cell Count / Q000187:drug effects; D002478:Cells, Cultured; D004298:Dopamine / Q000494:pharmacology*; D004958:Estradiol / Q000494:pharmacology*; D005260:Female; D019206:GTP-Binding Protein alpha Subunits, Gi-Go / Q000378:metabolism; D019205:GTP-Binding Protein alpha Subunits, Gs / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000378:metabolism*; D006462:Hemolytic Plaque Technique; D007150:Immunohistochemistry; D010052:Ovariectomy; D010903:Pituitary Gland, Anterior / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D011374:Progesterone / Q000494:pharmacology*; D011388:Prolactin / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D011374:Progesterone; D004958:Estradiol; D011388:Prolactin; D019204:GTP-Binding Proteins; D019206:GTP-Binding Protein alpha Subunits, Gi-Go; D019205:GTP-Binding Protein alpha Subunits, Gs; D004298:Dopamine