title: Phytohemagglutinin improves the development and ultrastructure of in vitro-cultured goat (Capra hircus) preantral follicles.
abstract: The objective this study was to determine the effect of phytohemagglutinin (PHA) on survival, growth and gene expression in caprine secondary follicles cultured in vitro. Secondary follicles (∼0.2 mm) were isolated from the cortex of caprine ovaries and cultured individually for 6 days in α-MEM+ supplemented with PHA (0, 1, 10, 50, 100, or 200 µg/mL). After 6 days of culture, follicle diameter and survival, antrum formation, ultrastructure and expression of mRNA for FSH receptors (FSH-R), proliferating cell nuclear antigen (PCNA), and neuronal nitric oxide synthase were determined. All treatments maintained follicular survival [α-MEM+ (94.59%); 1 µg/mL PHA (96.43%); 10 µg/mL PHA (84.85%); 50 µg/mL PHA (85.29%); 100 µg/mL PHA (88.57%), and 200 µg/mL PHA (87.50)], but the presence of 10 µg/mL PHA in the culture medium increased the antrum formation rate (21.21%) when compared with control (5.41%, P < 0.05) and ensured the maintenance of oocyte and granulosa cell ultrastructures after 6 days of culture. The expression of mRNA for FSH-R (2.7 ± 0.1) and PCNA (4.4 ± 0.2) was also significantly increased in follicles cultured with 10 µg/mL PHA in relation to those cultured in α-MEM+ (1.0 ± 0.1). In conclusion, supplementation of culture medium with 10 µg/mL PHA maintains the follicular viability and ultrastructure, and promotes the formation of antral cavity after 6 days of culture in vitro.
authors.affiliation: Núcleo de Biotecnologia de Sobral, NUBIS, Universidade Federal do Ceará, Sobral, CE, Brasil.
authors.forename: E V
authors.initials: EV
authors.lastname: Cunha
authors.forename: J J N
authors.initials: JJ
authors.lastname: Costa
authors.forename: R O D S
authors.initials: RO
authors.lastname: Rossi
authors.forename: A W B
authors.initials: AW
authors.lastname: Silva
authors.forename: J R S
authors.initials: JR
authors.lastname: Passos
authors.forename: A M L R
authors.initials: AM
authors.lastname: Portela
authors.forename: D C S T
authors.initials: DC
authors.lastname: Pereira
authors.forename: M A M
authors.initials: MA
authors.lastname: Donato
authors.forename: C C
authors.initials: CC
authors.lastname: Campello
authors.forename: M V A
authors.initials: MV
authors.lastname: Saraiva
authors.forename: C A
authors.initials: CA
authors.lastname: Peixoto
authors.forename: J R V
authors.initials: JR
authors.lastname: Silva
authors.forename: R P
authors.initials: RP
authors.lastname: Santos
mesh_terms: D000818:Animals; D005260:Female; D005640:Follicle Stimulating Hormone / Q000235:genetics / Q000378:metabolism*; D006041:Goats; D066298:In Vitro Techniques; D008934:Mitogens / Q000494:pharmacology*; D006080:Ovarian Follicle / Q000187:drug effects* / Q000254:growth & development / Q000648:ultrastructure; D010835:Phytohemagglutinins / Q000494:pharmacology*; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism
chemical_list: D008934:Mitogens; D010835:Phytohemagglutinins; D018809:Proliferating Cell Nuclear Antigen; D012333:RNA, Messenger; D005640:Follicle Stimulating Hormone
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