title: Expression of LH receptor mRNA splice variants in bovine granulosa cells: changes with follicle size and regulation by FSH in vitro.
abstract: In cattle, most evidence suggests that granulosa cells express LH receptors (LHR) after (or as) the follicle becomes dominant, however there is some suggestion that granulosa cells from smaller pre-dominant follicles may express several LHR mRNA splice variants. The objective of this study was to measure LHR expression in bovine follicles of defined size and steroidogenic ability, and in granulosa cells from small follicles (<6 mm diameter) undergoing differentiation in vitro. Semiquantitative RT-PCR demonstrated that LHR mRNA was undetectable in granulosa cells of follicles <7 mm diameter (nondominant follicles), and increased with follicle diameter in follicles >7 mm diameter. Splice variants with deletions of exon 10 and part of exon 11 were detected as previously described, and we detected a novel splice variant with a deletion of exon 3. Cultured granulosa cells contained LHR mRNA, but with significantly greater amounts of variants with deletions of exon 10 and/or exon 11 compared with cells from dominant follicles. FSH increased the abundance of some but not all LHR mRNA splice variants in cultured granulosa cells. The addition of LH to cultured cells did not increase progesterone secretion, despite the presence of LHR mRNA. Collectively, these data suggest that granulosa cells do not acquire functional LHR until follicle dominance occurs.
authors.affiliation: Departamento de Farmacologia, Instituto de Biociências, Universidade Estadual Paulista, Botucatu, São Paulo, Brazil. marcelo@assis.unesp.br
authors.forename: M F G
authors.initials: MF
authors.lastname: Nogueira
authors.forename: J
authors.initials: J
authors.lastname: Buratini
authors.forename: C A
authors.initials: CA
authors.lastname: Price
authors.forename: A C S
authors.initials: AC
authors.lastname: Castilho
authors.forename: M G L
authors.initials: MG
authors.lastname: Pinto
authors.forename: C M
authors.initials: CM
authors.lastname: Barros
mesh_terms: D017398:Alternative Splicing; D000818:Animals; D002417:Cattle / Q000235:genetics*; D048429:Cell Size; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D005260:Female; D005640:Follicle Stimulating Hormone / Q000494:pharmacology*; D006107:Granulosa Cells / Q000187:drug effects / Q000378:metabolism*; D006080:Ovarian Follicle / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger; D011974:Receptors, LH / Q000235:genetics* / Q000378:metabolism
chemical_list: D020033:Protein Isoforms; D012333:RNA, Messenger; D011974:Receptors, LH; D005640:Follicle Stimulating Hormone