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 culturedin 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.

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Main Authors: Cunha,E.V., Costa,J.J.N., Rossi,R.O.D.S., Silva,A.W.B., Passos,J.R.S., Portela,A.M.L.R., Pereira,D.C.S.T., Donato,M.A.M., Campello,C.C., Saraiva,M.V.A., Peixoto,C.A., Silva,J.R.V., Santos,R.P.
Format: Digital revista
Language:English
Published: Associação Brasileira de Divulgação Científica 2013
Online Access:http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2013000300245
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spelling oai:scielo:S0100-879X20130003002452015-10-08Phytohemagglutinin improves the development and ultrastructure of in vitro-cultured goat (Capra hircus) preantral folliclesCunha,E.V.Costa,J.J.N.Rossi,R.O.D.S.Silva,A.W.B.Passos,J.R.S.Portela,A.M.L.R.Pereira,D.C.S.T.Donato,M.A.M.Campello,C.C.Saraiva,M.V.A.Peixoto,C.A.Silva,J.R.V.Santos,R.P. Caprine Antrum formation FSH-R PCNA Ultrastructure The objective this study was to determine the effect of phytohemagglutinin (PHA) on survival, growth and gene expression in caprine secondary follicles culturedin 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.info:eu-repo/semantics/openAccessAssociação Brasileira de Divulgação CientíficaBrazilian Journal of Medical and Biological Research v.46 n.3 20132013-03-01info:eu-repo/semantics/articletext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2013000300245en10.1590/1414-431X20122702
institution SCIELO
collection OJS
country Brasil
countrycode BR
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access En linea
databasecode rev-scielo-br
tag revista
region America del Sur
libraryname SciELO
language English
format Digital
author Cunha,E.V.
Costa,J.J.N.
Rossi,R.O.D.S.
Silva,A.W.B.
Passos,J.R.S.
Portela,A.M.L.R.
Pereira,D.C.S.T.
Donato,M.A.M.
Campello,C.C.
Saraiva,M.V.A.
Peixoto,C.A.
Silva,J.R.V.
Santos,R.P.
spellingShingle Cunha,E.V.
Costa,J.J.N.
Rossi,R.O.D.S.
Silva,A.W.B.
Passos,J.R.S.
Portela,A.M.L.R.
Pereira,D.C.S.T.
Donato,M.A.M.
Campello,C.C.
Saraiva,M.V.A.
Peixoto,C.A.
Silva,J.R.V.
Santos,R.P.
Phytohemagglutinin improves the development and ultrastructure of in vitro-cultured goat (Capra hircus) preantral follicles
author_facet Cunha,E.V.
Costa,J.J.N.
Rossi,R.O.D.S.
Silva,A.W.B.
Passos,J.R.S.
Portela,A.M.L.R.
Pereira,D.C.S.T.
Donato,M.A.M.
Campello,C.C.
Saraiva,M.V.A.
Peixoto,C.A.
Silva,J.R.V.
Santos,R.P.
author_sort Cunha,E.V.
title 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_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_sort phytohemagglutinin improves the development and ultrastructure of in vitro-cultured goat (capra hircus) preantral follicles
description The objective this study was to determine the effect of phytohemagglutinin (PHA) on survival, growth and gene expression in caprine secondary follicles culturedin 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.
publisher Associação Brasileira de Divulgação Científica
publishDate 2013
url http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2013000300245
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