title: Effects of growth differentiation factor-9 and FSH on in vitro development, viability and mRNA expression in bovine preantral follicles.
abstract: The present study investigated the role of growth differentiation factor (GDF)-9 and FSH, alone or in combination, on the growth, viability and mRNA expression of FSH receptor, proliferating cell nuclear antigen (PCNA) and proteoglycan-related factors (i.e., hyaluronan synthase (HAS) 1, HAS2, versican, perlecan) in bovine secondary follicles before and after in vitro culture. After 12 days culture, sequential FSH (100 ng mL⁻¹) from Days 0 to 6 and 500 ng mL⁻¹ from Days 7 to 12) increased follicular diameter and resulted in increased antrum formation (P<0.05). Alone, 200 ng mL⁻¹ GDF-9 significantly reduced HAS1 mRNA levels, but increased versican and perlecan mRNA levels in whole follicles, which included the oocyte, theca and granulosa cells. Together, FSH and GDF-9 increased HAS2 and versican (VCAN) mRNA levels, but decreased PCNA mRNA expression, compared with levels in follicles cultured in α-minimum essential medium supplemented with 3.0 mg mL⁻¹ bovine serum albumin, 10 µg mL⁻¹ insulin, 5.5 µg mL⁻¹ transferrin, 5 ng mL⁻¹ selenium, 2 mM glutamine, 2mM hypoxanthine and 50 μg mL⁻¹ ascorbic acid (α-MEM⁺). Comparisons of uncultured (0.2 mm) and α-MEM⁺ cultured follicles revealed that HAS1 mRNA expression was higher, whereas VCAN expression was lower, in cultured follicles (P<0.05). Expression of HAS1, VCAN and perlecan (HSPG2) was higher in cultured than in vivo-grown (0.3 mm) follicles. In conclusion, FSH and/or GDF-9 promote follicular growth and antrum formation. Moreover, GDF-9 stimulates expression of versican and perlecan and interacts positively with FSH to increase HAS2 expression.
authors.affiliation: Biotechnology Nucleus of Sobral (NUBIS), Federal University of Ceara, CEP 62042-280, Sobral, CE, Brazil.
authors.forename: G L
authors.initials: GL
authors.lastname: Vasconcelos
authors.forename: M V A
authors.initials: MV
authors.lastname: Saraiva
authors.forename: J J N
authors.initials: JJ
authors.lastname: Costa
authors.forename: M J
authors.initials: MJ
authors.lastname: Passos
authors.forename: A W B
authors.initials: AW
authors.lastname: Silva
authors.forename: R O D S
authors.initials: RO
authors.lastname: Rossi
authors.forename: A M L R
authors.initials: AM
authors.lastname: Portela
authors.forename: A B G
authors.initials: AB
authors.lastname: Duarte
authors.forename: D M
authors.initials: DM
authors.lastname: Magalhães-Padilha
authors.forename: C C
authors.initials: CC
authors.lastname: Campelo
authors.forename: J R
authors.initials: JR
authors.lastname: Figueiredo
authors.forename: R
authors.initials: R
authors.lastname: van den Hurk
authors.forename: J R V
authors.initials: JR
authors.lastname: Silva
mesh_terms: D000003:Abattoirs; D000818:Animals; D002417:Cattle; D002470:Cell Survival; D005260:Female; D005640:Follicle Stimulating Hormone / Q000378:metabolism*; D015571:Follicular Fluid / Q000201:enzymology / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D014453:Glucuronosyltransferase / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D055429:Growth Differentiation Factor 9 / Q000378:metabolism*; D000076002:Hyaluronan Synthases; D059471:In Vitro Oocyte Maturation Techniques / Q000662:veterinary*; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D009865:Oocytes / Q000166:cytology / Q000201:enzymology / Q000378:metabolism; D009866:Oogenesis; D006080:Ovarian Follicle / Q000166:cytology / Q000254:growth & development / Q000378:metabolism*; D018809:Proliferating Cell Nuclear Antigen / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011509:Proteoglycans / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D011962:Receptors, FSH / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D046509:Tissue Culture Techniques / Q000662:veterinary
chemical_list: D055429:Growth Differentiation Factor 9; D007527:Isoenzymes; D018809:Proliferating Cell Nuclear Antigen; D011509:Proteoglycans; D012333:RNA, Messenger; D011962:Receptors, FSH; D005640:Follicle Stimulating Hormone; D014453:Glucuronosyltransferase; D000076002:Hyaluronan Synthases