title: Effects of angiotensin II, atrial natriuretic peptide and endothelin-1 on proliferation and steroidogenic output of bovine granulosa cells cultured in a chemically defined system.
abstract: The role of local factors in the modulation of granulosa cell (GC) proliferation and differentiation is well described in the literature. The present work used a long-term bovine GC culture, in chemically defined medium without gonadotropins, to study the effects of angiotensin II (Ang II), atrial natriuretic peptide (ANP) and endothelin-1 (EDN1) on the steroidogenesis and cellular proliferation. Small follicles (3-5mm in diameter) from ovaries obtained in the slaughterhouse were selected according to their vascularization and follicular fluid color in order to isolate GC. Granulosa cells were plated at a density of 5×10(4)cells/well in supplemented alpha-MEM containing 3 levels (0, 10(-8)M and 10(-7)M) of Ang II, ANP, and EDN1 for up to 96h. Proliferation was evaluated by tritiated thymidine incorporation. The results showed that Ang II, ANP, and EDN1 modulate the steroidogenic output and proliferation index of GCs depending on the dose and time of culture. The selected vasoactive peptides increased androstenedione (A4) consumption in parallel with increased estradiol (E2). Although the peptides also promoted a significant increase in pregnenolone (P5) and progesterone (P4) production, the E2:P4 ratio was maintained at a high at most of the tested doses. Taken together, our in vitro data suggest that these vasoactive factors may have a direct effect on physiological follicular deviation, favoring dominance of the selected follicle.
authors.affiliation: Department of Physiology, School of Medicine of Ribeirão Preto, University of São Paulo, SP, Brazil; Department of Biological Science and Health, University Center of Araraquara, UNIARA, Araraquara, SP, Brazil. Electronic address: luis.montrezor@icloud.com.
authors.forename: L H
authors.initials: LH
authors.lastname: Montrezor
authors.affiliation: Department of Physiology, School of Medicine of Ribeirão Preto, University of São Paulo, SP, Brazil; Hospital Israelita Albert Einstein, São Paulo, SP, Brazil.
authors.forename: C A
authors.initials: CA
authors.lastname: Piccinato
authors.affiliation: Department of Physiology, School of Medicine of Ribeirão Preto, University of São Paulo, SP, Brazil.
authors.forename: C V A
authors.initials: CV
authors.lastname: Collares
authors.affiliation: Department of Physiology, School of Medicine of Ribeirão Preto, University of São Paulo, SP, Brazil.
authors.forename: A A
authors.initials: AA
authors.lastname: Vireque
authors.affiliation: Department of Physiology, School of Medicine of Ribeirão Preto, University of São Paulo, SP, Brazil; Laboratory for the Study of Reproduction, CSF, Biological Institute, University of Brasília, Brasília, DF, Brazil.
authors.forename: A A M Rosa E
authors.initials: AA
authors.lastname: Silva
mesh_terms: D000804:Angiotensin II / Q000494:pharmacology*; D000818:Animals; D009320:Atrial Natriuretic Factor / Q000494:pharmacology*; D002417:Cattle; D018929:Cell Culture Techniques / Q000662:veterinary; D049109:Cell Proliferation / Q000187:drug effects*; D002478:Cells, Cultured; D019332:Endothelin-1 / Q000494:pharmacology*; D005260:Female; D006107:Granulosa Cells / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D013256:Steroids / Q000378:metabolism
chemical_list: D019332:Endothelin-1; D013256:Steroids; D000804:Angiotensin II; D009320:Atrial Natriuretic Factor
keywords: Angiotensin II; Atrial natriuretic peptide; Bovine; Endothelin-1; Granulosa cells; Steroidogenesis