Phytohemagglutinin improves the development and ultrastructure of in vitro-cultured goat (Capra hircus) preantral follicles

• Main Authors: E.V. Cunha, J.J.N. Costa, R.O.D.S. Rossi, A.W.B. Silva, J.R.S. Passos, A.M.L.R. Portela, D.C.S.T. Pereira, M.A.M. Donato, C.C. Campello, M.V.A. Saraiva, C.A. Peixoto, J.R.V. Silva, R.P. Santos
• Format: Article
• Language: English
• Published: Associação Brasileira de Divulgação Científica 2013-03-01
• Series: Brazilian Journal of Medical and Biological Research
• Subjects: Caprine; Antrum formation; FSH-R; PCNA; Ultrastructure
• Online Access: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2013000300245&lng=en&tlng=en

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.