Phytohemagglutinin improves the development and ultrastructure of in vitro-cultured goat (Capra hircus) preantral follicles
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...
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| 语言: | English |
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Associação Brasileira de Divulgação Científica
2013-01-01
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| 丛编: | Brazilian Journal of Medical and Biological Research |
| 主题: | |
| 在线阅读: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2013000300245 |
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| author | 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 |
| author_facet | 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 |
| author_sort | E.V. Cunha |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-11T08:38:57Z |
| format | Article |
| id | doaj.art-3bacc254d81c47548ca142da95fb42dc |
| institution | Directory Open Access Journal |
| issn | 0100-879X 1414-431X |
| language | English |
| last_indexed | 2024-12-11T08:38:57Z |
| publishDate | 2013-01-01 |
| publisher | Associação Brasileira de Divulgação Científica |
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| series | Brazilian Journal of Medical and Biological Research |
| spelling | doaj.art-3bacc254d81c47548ca142da95fb42dc2022-12-22T01:14:17ZengAssociação Brasileira de Divulgação CientíficaBrazilian Journal of Medical and Biological Research0100-879X1414-431X2013-01-01463245252Phytohemagglutinin improves the development and ultrastructure of in vitro-cultured goat (Capra hircus) preantral folliclesE.V. CunhaJ.J.N. CostaR.O.D.S. RossiA.W.B. SilvaJ.R.S. PassosA.M.L.R. PortelaD.C.S.T. PereiraM.A.M. DonatoC.C. CampelloM.V.A. SaraivaC.A. PeixotoJ.R.V. SilvaR.P. SantosThe 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.http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2013000300245CaprineAntrum formationFSH-RPCNAUltrastructure |
| spellingShingle | 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 Phytohemagglutinin improves the development and ultrastructure of in vitro-cultured goat (Capra hircus) preantral follicles Brazilian Journal of Medical and Biological Research Caprine Antrum formation FSH-R PCNA Ultrastructure |
| title | Phytohemagglutinin improves the development and ultrastructure of in vitro-cultured goat (Capra hircus) preantral follicles |
| title_full | Phytohemagglutinin improves the development and ultrastructure of in vitro-cultured goat (Capra hircus) preantral follicles |
| title_fullStr | Phytohemagglutinin improves the development and ultrastructure of in vitro-cultured goat (Capra hircus) preantral follicles |
| title_full_unstemmed | Phytohemagglutinin improves the development and ultrastructure of in vitro-cultured goat (Capra hircus) preantral follicles |
| title_short | Phytohemagglutinin improves the development and ultrastructure of in vitro-cultured goat (Capra hircus) preantral follicles |
| title_sort | phytohemagglutinin improves the development and ultrastructure of in vitro cultured goat capra hircus preantral follicles |
| topic | Caprine Antrum formation FSH-R PCNA Ultrastructure |
| url | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2013000300245 |
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