Differential effects of activin-A and FSH on growth, viability and messenger RNA expression in cultured bovine preantral follicles

A. W B Silva; F. T G Bezerra; J. J N Costa; R. O D S Rossi; M. J. Passos; G. L. Vasconcelos; R. Rossetto; M. A M Donato; D. M. Magalhães-Padilha; C. C. Campello; M. V A Saraiva; J. R. Figueiredo; C. A. Peixoto; R. Van den Hurk; J. R V Silva

doi:10.1016/j.livsci.2013.12.003

Differential effects of activin-A and FSH on growth, viability and messenger RNA expression in cultured bovine preantral follicles

A. W B Silva, F. T G Bezerra, J. J N Costa, R. O D S Rossi, M. J. Passos, G. L. Vasconcelos, R. Rossetto, M. A M Donato, D. M. Magalhães-Padilha, C. C. Campello, M. V A Saraiva, J. R. Figueiredo, C. A. Peixoto, R. Van den Hurk, J. R V Silva^*

^*Corresponding author for this work

Veterinary Medicine

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

This study evaluated the effect of follicle stimulating hormone (FSH) alone or in combination with activin-A on survival, growth and expression of mRNA for follicle stimulating hormone receptor (FSH-R), proliferating cell nuclear antigen (PCNA), activin receptor type I B (ActR-IB), activin receptor type II B (ActR-IIB), hyaluronan synthase-1 (HAS 1) and hyaluronan synthase-2 (HAS 2) in bovine secondary follicles cultured in vitro for 18 days. Preantral follicles (~0.2mm) were isolated and individually cultured in absence (α-minimum essential medium (α-MEM⁺): control) or presence of activin-A, FSH in increasing concentrations or both activin-A and FSH. An increase in follicular diameter was observed after 6 days of culture in all culture treatments compared to control medium; after 12 days in treatment with FSH alone or the mixture of FSH and activin-A (P<0.05); and, after 18 days only in the presence of FSH alone (P<0.05). However, in combination with activin-A, FSH-stimulated follicle growth after 18 days was blocked (P<0.05) and, had significantly lower (P<0.05) levels of mRNA for ActR-IB, ActR-IIB, when compared to α-MEM⁺, and for FSH-R and PCNA, when compared to α-MEM⁺ supplemented with activin-A only. Moreover^, follicles cultured in presence of FSH alone had greater (P<0.05) levels of mRNA for HAS 1 and HAS 2 than those cultured in medium supplemented with both activin-A and FSH. Morphological, immunofluorescence and ultrastructural analysis confirmed the integrity of follicles cultured in FSH after 18 days. In conclusion, activin-A exerts a stimulatory effect on in vitro early follicular development for up to 6 days, has no effect after 12 days of culture and blocks the stimulatory growth effect of FSH after 18 days. The reduced mRNA levels in follicles cultured with both activin-A and FSH suggest a decreased sensitivity of follicles for activin-A and FSH and inhibited follicular cell proliferation after long-term in vitro culture of isolated preantral follicles.

Original language	English
Pages (from-to)	199-207
Number of pages	9
Journal	Livestock Science
Volume	160
Issue number	1
DOIs	https://doi.org/10.1016/j.livsci.2013.12.003
Publication status	Published - 1 Jan 2014

Keywords

Activin-A
Bovine
FSH
Messenger RNA expression
Preantral follicles
Ultrastructure

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10.1016/j.livsci.2013.12.003

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Silva, A. W. B., Bezerra, F. T. G., Costa, J. J. N., Rossi, R. O. D. S., Passos, M. J., Vasconcelos, G. L., Rossetto, R., Donato, M. A. M., Magalhães-Padilha, D. M., Campello, C. C., Saraiva, M. V. A., Figueiredo, J. R., Peixoto, C. A., Van den Hurk, R., & Silva, J. R. V. (2014). Differential effects of activin-A and FSH on growth, viability and messenger RNA expression in cultured bovine preantral follicles. Livestock Science, 160(1), 199-207. https://doi.org/10.1016/j.livsci.2013.12.003

@article{9bc5ec153a55437da3161227c9cc5f20,

title = "Differential effects of activin-A and FSH on growth, viability and messenger RNA expression in cultured bovine preantral follicles",

abstract = "This study evaluated the effect of follicle stimulating hormone (FSH) alone or in combination with activin-A on survival, growth and expression of mRNA for follicle stimulating hormone receptor (FSH-R), proliferating cell nuclear antigen (PCNA), activin receptor type I B (ActR-IB), activin receptor type II B (ActR-IIB), hyaluronan synthase-1 (HAS 1) and hyaluronan synthase-2 (HAS 2) in bovine secondary follicles cultured in vitro for 18 days. Preantral follicles (~0.2mm) were isolated and individually cultured in absence (α-minimum essential medium (α-MEM+): control) or presence of activin-A, FSH in increasing concentrations or both activin-A and FSH. An increase in follicular diameter was observed after 6 days of culture in all culture treatments compared to control medium; after 12 days in treatment with FSH alone or the mixture of FSH and activin-A (P<0.05); and, after 18 days only in the presence of FSH alone (P<0.05). However, in combination with activin-A, FSH-stimulated follicle growth after 18 days was blocked (P<0.05) and, had significantly lower (P<0.05) levels of mRNA for ActR-IB, ActR-IIB, when compared to α-MEM+, and for FSH-R and PCNA, when compared to α-MEM+ supplemented with activin-A only. Moreover, follicles cultured in presence of FSH alone had greater (P<0.05) levels of mRNA for HAS 1 and HAS 2 than those cultured in medium supplemented with both activin-A and FSH. Morphological, immunofluorescence and ultrastructural analysis confirmed the integrity of follicles cultured in FSH after 18 days. In conclusion, activin-A exerts a stimulatory effect on in vitro early follicular development for up to 6 days, has no effect after 12 days of culture and blocks the stimulatory growth effect of FSH after 18 days. The reduced mRNA levels in follicles cultured with both activin-A and FSH suggest a decreased sensitivity of follicles for activin-A and FSH and inhibited follicular cell proliferation after long-term in vitro culture of isolated preantral follicles.",

keywords = "Activin-A, Bovine, FSH, Messenger RNA expression, Preantral follicles, Ultrastructure",

author = "Silva, {A. W B} and Bezerra, {F. T G} and Costa, {J. J N} and Rossi, {R. O D S} and Passos, {M. J.} and Vasconcelos, {G. L.} and R. Rossetto and Donato, {M. A M} and Magalh{\~a}es-Padilha, {D. M.} and Campello, {C. C.} and Saraiva, {M. V A} and Figueiredo, {J. R.} and Peixoto, {C. A.} and {Van den Hurk}, R. and Silva, {J. R V}",

year = "2014",

month = jan,

day = "1",

doi = "10.1016/j.livsci.2013.12.003",

language = "English",

volume = "160",

pages = "199--207",

journal = "Livestock Science",

issn = "1871-1413",

publisher = "Elsevier BV",

number = "1",

}

Silva, AWB, Bezerra, FTG, Costa, JJN, Rossi, RODS, Passos, MJ, Vasconcelos, GL, Rossetto, R, Donato, MAM, Magalhães-Padilha, DM, Campello, CC, Saraiva, MVA, Figueiredo, JR, Peixoto, CA, Van den Hurk, R & Silva, JRV 2014, 'Differential effects of activin-A and FSH on growth, viability and messenger RNA expression in cultured bovine preantral follicles', Livestock Science, vol. 160, no. 1, pp. 199-207. https://doi.org/10.1016/j.livsci.2013.12.003

TY - JOUR

T1 - Differential effects of activin-A and FSH on growth, viability and messenger RNA expression in cultured bovine preantral follicles

AU - Silva, A. W B

AU - Bezerra, F. T G

AU - Costa, J. J N

AU - Rossi, R. O D S

AU - Passos, M. J.

AU - Vasconcelos, G. L.

AU - Rossetto, R.

AU - Donato, M. A M

AU - Magalhães-Padilha, D. M.

AU - Campello, C. C.

AU - Saraiva, M. V A

AU - Figueiredo, J. R.

AU - Peixoto, C. A.

AU - Van den Hurk, R.

AU - Silva, J. R V

PY - 2014/1/1

Y1 - 2014/1/1

N2 - This study evaluated the effect of follicle stimulating hormone (FSH) alone or in combination with activin-A on survival, growth and expression of mRNA for follicle stimulating hormone receptor (FSH-R), proliferating cell nuclear antigen (PCNA), activin receptor type I B (ActR-IB), activin receptor type II B (ActR-IIB), hyaluronan synthase-1 (HAS 1) and hyaluronan synthase-2 (HAS 2) in bovine secondary follicles cultured in vitro for 18 days. Preantral follicles (~0.2mm) were isolated and individually cultured in absence (α-minimum essential medium (α-MEM+): control) or presence of activin-A, FSH in increasing concentrations or both activin-A and FSH. An increase in follicular diameter was observed after 6 days of culture in all culture treatments compared to control medium; after 12 days in treatment with FSH alone or the mixture of FSH and activin-A (P<0.05); and, after 18 days only in the presence of FSH alone (P<0.05). However, in combination with activin-A, FSH-stimulated follicle growth after 18 days was blocked (P<0.05) and, had significantly lower (P<0.05) levels of mRNA for ActR-IB, ActR-IIB, when compared to α-MEM+, and for FSH-R and PCNA, when compared to α-MEM+ supplemented with activin-A only. Moreover, follicles cultured in presence of FSH alone had greater (P<0.05) levels of mRNA for HAS 1 and HAS 2 than those cultured in medium supplemented with both activin-A and FSH. Morphological, immunofluorescence and ultrastructural analysis confirmed the integrity of follicles cultured in FSH after 18 days. In conclusion, activin-A exerts a stimulatory effect on in vitro early follicular development for up to 6 days, has no effect after 12 days of culture and blocks the stimulatory growth effect of FSH after 18 days. The reduced mRNA levels in follicles cultured with both activin-A and FSH suggest a decreased sensitivity of follicles for activin-A and FSH and inhibited follicular cell proliferation after long-term in vitro culture of isolated preantral follicles.

AB - This study evaluated the effect of follicle stimulating hormone (FSH) alone or in combination with activin-A on survival, growth and expression of mRNA for follicle stimulating hormone receptor (FSH-R), proliferating cell nuclear antigen (PCNA), activin receptor type I B (ActR-IB), activin receptor type II B (ActR-IIB), hyaluronan synthase-1 (HAS 1) and hyaluronan synthase-2 (HAS 2) in bovine secondary follicles cultured in vitro for 18 days. Preantral follicles (~0.2mm) were isolated and individually cultured in absence (α-minimum essential medium (α-MEM+): control) or presence of activin-A, FSH in increasing concentrations or both activin-A and FSH. An increase in follicular diameter was observed after 6 days of culture in all culture treatments compared to control medium; after 12 days in treatment with FSH alone or the mixture of FSH and activin-A (P<0.05); and, after 18 days only in the presence of FSH alone (P<0.05). However, in combination with activin-A, FSH-stimulated follicle growth after 18 days was blocked (P<0.05) and, had significantly lower (P<0.05) levels of mRNA for ActR-IB, ActR-IIB, when compared to α-MEM+, and for FSH-R and PCNA, when compared to α-MEM+ supplemented with activin-A only. Moreover, follicles cultured in presence of FSH alone had greater (P<0.05) levels of mRNA for HAS 1 and HAS 2 than those cultured in medium supplemented with both activin-A and FSH. Morphological, immunofluorescence and ultrastructural analysis confirmed the integrity of follicles cultured in FSH after 18 days. In conclusion, activin-A exerts a stimulatory effect on in vitro early follicular development for up to 6 days, has no effect after 12 days of culture and blocks the stimulatory growth effect of FSH after 18 days. The reduced mRNA levels in follicles cultured with both activin-A and FSH suggest a decreased sensitivity of follicles for activin-A and FSH and inhibited follicular cell proliferation after long-term in vitro culture of isolated preantral follicles.

KW - Activin-A

KW - Bovine

KW - FSH

KW - Messenger RNA expression

KW - Preantral follicles

KW - Ultrastructure

UR - http://www.scopus.com/inward/record.url?scp=84892536107&partnerID=8YFLogxK

U2 - 10.1016/j.livsci.2013.12.003

DO - 10.1016/j.livsci.2013.12.003

M3 - Article

AN - SCOPUS:84892536107

SN - 1871-1413

VL - 160

SP - 199

EP - 207

JO - Livestock Science

JF - Livestock Science

IS - 1

ER -

Differential effects of activin-A and FSH on growth, viability and messenger RNA expression in cultured bovine preantral follicles

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Keywords

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