FSH dose and strategy of administration during ovarian stimulation alter the gene expression profile in ovine cumulus-oocyte complexes.
Ovarian stimulation is an important tool to increase the number of oocytes obtained by laparoscopy for the in vitro production of embryos (IVP). In sheep, different concentrations of FSH administered in single dose (SD) or multiple doses (MD) have been adopted. In parallel, the oocyte quality is fundamental for IVP success, so strategies to produce more competent oocytes have been evaluated. The aim of this study was to evaluate the gene expression profile of BCB+ COC from different hormonal protocols of ovarian stimulation in Santa Inês ewes. To achieve that, a cross-over design was used, where 12 pluriparous ewes had their follicular wave synchronized (Balaro et al., Domest Anim Endocrinol, 54: 10-14, 2016). At 80 h after progestogen implant removal, all ewes received a new vaginal sponge and it started the stimulation by administration of: 80 (Group 1 - 80-SD) or 120 (Group 2 - 120-SD) mg FSH (Folltropin-V®, Bioniche Animal Health, Ontario, Canada) and 300 IU of eCG both in single dose, or 80 (Group 3 - 80-MD) or 120 (Group 4 - 120-MD) in decreasing doses (50/30/20%) every 12 h. The COCs were recovered by laparoscopy and classified morphologically in grade I / II (homogeneous ooplasm and more than 3 cumulus cells layers), III (homogeneous ooplasm and less than 3 cumulus cells layers or partially denuded) and IV (heterogeneous ooplasm or degenerate). For inference of the development competence GI, II and III COCs were stained with bright cresyl blue (BCB) and classified into: BCB+ (competent) and BCB- (non-competent). These variables were compared by ANOVA followed by Tukey test. The abundance of mRNA that encodes proteins associated with steroidogenesis (STAR, FSHr, LHr and ER?), oocyte quality (MATER, BMP15, GDF9 and ZAR1) and apoptosis (BAX and Bcl-2) was assessed by real-time qPCR normalized with GAPDH in BCB+ COCs. The abundance of gene transcripts associated with steroidogenesis was down-regulated (P <0.05) with increasing FSH concentration, when administration was performed in a single dose (80-SD and 120-SD). On the other hand, when the administration was performed in MD, only the LHr was down-regulated (80-MD and 120-MD). In the 80-MD group, FSHr and Er? were down-regulated (P <0.05) in comparison with 80-SD. For genes related with oocyte quality, 80-MD showed up-regulation (P <0.05) to MATER (when compared to 80-SD), ZAR1 and MATER (compared to 120-SD). Nonetheless, apoptosis genes were not affected. These data demonstrate that the FSH dose and strategy of administration affect the gene expression profile in ovine COCs. Subsequent studies are necessary to assess the effect of this change on maturation rate and developmental competence.
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2017-08-23
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Subjects: | Expressão gênica, Produção de embriões in vitro, Embryo Technology, In vitro embryo production., FSH, Ovino, Reprodução animal, Transferência de embrião, Sheep, Reproduction, Ova, Gene expression., |
Online Access: | http://www.alice.cnptia.embrapa.br/alice/handle/doc/1074359 |
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dig-alice-doc-10743592017-08-26T04:03:28Z FSH dose and strategy of administration during ovarian stimulation alter the gene expression profile in ovine cumulus-oocyte complexes. BRAGANÇA, G. M. BATISTA, R. I. T. P. SOUZA FABJAN, J. M. G. de ALFRADIQUE, V. A. P. CORTÊS, L. R. CAMARGO, L. S. de A. FONSECA, J. F. da BRANDÃO, F. Z. Gláucia Mota Bragança, Universidade Federal Fluminense (UFF) - Rio de Janeiro, RJ; Ribrio Ivan Tavares Pereira Batista; Joanna Maria Gonçalves de Souza Fabjan; Vivian Angélico Pereira Alfradique; Luana Rangel Côrtes; LUIZ SERGIO DE ALMEIDA CAMARGO, CNPGL; JEFERSON FERREIRA DA FONSECA, CNPC; Felipe Zandonadi Brandao. Expressão gênica Produção de embriões in vitro Embryo Technology In vitro embryo production. FSH Ovino Reprodução animal Transferência de embrião Sheep Reproduction Ova Gene expression. Ovarian stimulation is an important tool to increase the number of oocytes obtained by laparoscopy for the in vitro production of embryos (IVP). In sheep, different concentrations of FSH administered in single dose (SD) or multiple doses (MD) have been adopted. In parallel, the oocyte quality is fundamental for IVP success, so strategies to produce more competent oocytes have been evaluated. The aim of this study was to evaluate the gene expression profile of BCB+ COC from different hormonal protocols of ovarian stimulation in Santa Inês ewes. To achieve that, a cross-over design was used, where 12 pluriparous ewes had their follicular wave synchronized (Balaro et al., Domest Anim Endocrinol, 54: 10-14, 2016). At 80 h after progestogen implant removal, all ewes received a new vaginal sponge and it started the stimulation by administration of: 80 (Group 1 - 80-SD) or 120 (Group 2 - 120-SD) mg FSH (Folltropin-V®, Bioniche Animal Health, Ontario, Canada) and 300 IU of eCG both in single dose, or 80 (Group 3 - 80-MD) or 120 (Group 4 - 120-MD) in decreasing doses (50/30/20%) every 12 h. The COCs were recovered by laparoscopy and classified morphologically in grade I / II (homogeneous ooplasm and more than 3 cumulus cells layers), III (homogeneous ooplasm and less than 3 cumulus cells layers or partially denuded) and IV (heterogeneous ooplasm or degenerate). For inference of the development competence GI, II and III COCs were stained with bright cresyl blue (BCB) and classified into: BCB+ (competent) and BCB- (non-competent). These variables were compared by ANOVA followed by Tukey test. The abundance of mRNA that encodes proteins associated with steroidogenesis (STAR, FSHr, LHr and ER?), oocyte quality (MATER, BMP15, GDF9 and ZAR1) and apoptosis (BAX and Bcl-2) was assessed by real-time qPCR normalized with GAPDH in BCB+ COCs. The abundance of gene transcripts associated with steroidogenesis was down-regulated (P <0.05) with increasing FSH concentration, when administration was performed in a single dose (80-SD and 120-SD). On the other hand, when the administration was performed in MD, only the LHr was down-regulated (80-MD and 120-MD). In the 80-MD group, FSHr and Er? were down-regulated (P <0.05) in comparison with 80-SD. For genes related with oocyte quality, 80-MD showed up-regulation (P <0.05) to MATER (when compared to 80-SD), ZAR1 and MATER (compared to 120-SD). Nonetheless, apoptosis genes were not affected. These data demonstrate that the FSH dose and strategy of administration affect the gene expression profile in ovine COCs. Subsequent studies are necessary to assess the effect of this change on maturation rate and developmental competence. Proceedings of the 31st Annual Meeting of the Brazilian Embryo Technology Society (SBTE), Cabo de Santo Agostinho, PE, Brazil, August 17 to 19, 2017. 2017-08-25T19:11:50Z 2017-08-25T19:11:50Z 2017-08-23 2017 2019-09-23T11:11:11Z Anais e Proceedings de eventos Animal Reprodroduction, v. 14, n. 3, p. 746, Jul./Sept. 2017. http://www.alice.cnptia.embrapa.br/alice/handle/doc/1074359 en eng openAccess |
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Expressão gênica Produção de embriões in vitro Embryo Technology In vitro embryo production. FSH Ovino Reprodução animal Transferência de embrião Sheep Reproduction Ova Gene expression. Expressão gênica Produção de embriões in vitro Embryo Technology In vitro embryo production. FSH Ovino Reprodução animal Transferência de embrião Sheep Reproduction Ova Gene expression. |
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Expressão gênica Produção de embriões in vitro Embryo Technology In vitro embryo production. FSH Ovino Reprodução animal Transferência de embrião Sheep Reproduction Ova Gene expression. Expressão gênica Produção de embriões in vitro Embryo Technology In vitro embryo production. FSH Ovino Reprodução animal Transferência de embrião Sheep Reproduction Ova Gene expression. BRAGANÇA, G. M. BATISTA, R. I. T. P. SOUZA FABJAN, J. M. G. de ALFRADIQUE, V. A. P. CORTÊS, L. R. CAMARGO, L. S. de A. FONSECA, J. F. da BRANDÃO, F. Z. FSH dose and strategy of administration during ovarian stimulation alter the gene expression profile in ovine cumulus-oocyte complexes. |
description |
Ovarian stimulation is an important tool to increase the number of oocytes obtained by laparoscopy for the in vitro production of embryos (IVP). In sheep, different concentrations of FSH administered in single dose (SD) or multiple doses (MD) have been adopted. In parallel, the oocyte quality is fundamental for IVP success, so strategies to produce more competent oocytes have been evaluated. The aim of this study was to evaluate the gene expression profile of BCB+ COC from different hormonal protocols of ovarian stimulation in Santa Inês ewes. To achieve that, a cross-over design was used, where 12 pluriparous ewes had their follicular wave synchronized (Balaro et al., Domest Anim Endocrinol, 54: 10-14, 2016). At 80 h after progestogen implant removal, all ewes received a new vaginal sponge and it started the stimulation by administration of: 80 (Group 1 - 80-SD) or 120 (Group 2 - 120-SD) mg FSH (Folltropin-V®, Bioniche Animal Health, Ontario, Canada) and 300 IU of eCG both in single dose, or 80 (Group 3 - 80-MD) or 120 (Group 4 - 120-MD) in decreasing doses (50/30/20%) every 12 h. The COCs were recovered by laparoscopy and classified morphologically in grade I / II (homogeneous ooplasm and more than 3 cumulus cells layers), III (homogeneous ooplasm and less than 3 cumulus cells layers or partially denuded) and IV (heterogeneous ooplasm or degenerate). For inference of the development competence GI, II and III COCs were stained with bright cresyl blue (BCB) and classified into: BCB+ (competent) and BCB- (non-competent). These variables were compared by ANOVA followed by Tukey test. The abundance of mRNA that encodes proteins associated with steroidogenesis (STAR, FSHr, LHr and ER?), oocyte quality (MATER, BMP15, GDF9 and ZAR1) and apoptosis (BAX and Bcl-2) was assessed by real-time qPCR normalized with GAPDH in BCB+ COCs. The abundance of gene transcripts associated with steroidogenesis was down-regulated (P <0.05) with increasing FSH concentration, when administration was performed in a single dose (80-SD and 120-SD). On the other hand, when the administration was performed in MD, only the LHr was down-regulated (80-MD and 120-MD). In the 80-MD group, FSHr and Er? were down-regulated (P <0.05) in comparison with 80-SD. For genes related with oocyte quality, 80-MD showed up-regulation (P <0.05) to MATER (when compared to 80-SD), ZAR1 and MATER (compared to 120-SD). Nonetheless, apoptosis genes were not affected. These data demonstrate that the FSH dose and strategy of administration affect the gene expression profile in ovine COCs. Subsequent studies are necessary to assess the effect of this change on maturation rate and developmental competence. |
author2 |
Gláucia Mota Bragança, Universidade Federal Fluminense (UFF) - Rio de Janeiro, RJ; Ribrio Ivan Tavares Pereira Batista; Joanna Maria Gonçalves de Souza Fabjan; Vivian Angélico Pereira Alfradique; Luana Rangel Côrtes; LUIZ SERGIO DE ALMEIDA CAMARGO, CNPGL; JEFERSON FERREIRA DA FONSECA, CNPC; Felipe Zandonadi Brandao. |
author_facet |
Gláucia Mota Bragança, Universidade Federal Fluminense (UFF) - Rio de Janeiro, RJ; Ribrio Ivan Tavares Pereira Batista; Joanna Maria Gonçalves de Souza Fabjan; Vivian Angélico Pereira Alfradique; Luana Rangel Côrtes; LUIZ SERGIO DE ALMEIDA CAMARGO, CNPGL; JEFERSON FERREIRA DA FONSECA, CNPC; Felipe Zandonadi Brandao. BRAGANÇA, G. M. BATISTA, R. I. T. P. SOUZA FABJAN, J. M. G. de ALFRADIQUE, V. A. P. CORTÊS, L. R. CAMARGO, L. S. de A. FONSECA, J. F. da BRANDÃO, F. Z. |
format |
Anais e Proceedings de eventos |
topic_facet |
Expressão gênica Produção de embriões in vitro Embryo Technology In vitro embryo production. FSH Ovino Reprodução animal Transferência de embrião Sheep Reproduction Ova Gene expression. |
author |
BRAGANÇA, G. M. BATISTA, R. I. T. P. SOUZA FABJAN, J. M. G. de ALFRADIQUE, V. A. P. CORTÊS, L. R. CAMARGO, L. S. de A. FONSECA, J. F. da BRANDÃO, F. Z. |
author_sort |
BRAGANÇA, G. M. |
title |
FSH dose and strategy of administration during ovarian stimulation alter the gene expression profile in ovine cumulus-oocyte complexes. |
title_short |
FSH dose and strategy of administration during ovarian stimulation alter the gene expression profile in ovine cumulus-oocyte complexes. |
title_full |
FSH dose and strategy of administration during ovarian stimulation alter the gene expression profile in ovine cumulus-oocyte complexes. |
title_fullStr |
FSH dose and strategy of administration during ovarian stimulation alter the gene expression profile in ovine cumulus-oocyte complexes. |
title_full_unstemmed |
FSH dose and strategy of administration during ovarian stimulation alter the gene expression profile in ovine cumulus-oocyte complexes. |
title_sort |
fsh dose and strategy of administration during ovarian stimulation alter the gene expression profile in ovine cumulus-oocyte complexes. |
publishDate |
2017-08-23 |
url |
http://www.alice.cnptia.embrapa.br/alice/handle/doc/1074359 |
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