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Registro Completo |
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Biblioteca(s): |
Embrapa Gado de Leite; Embrapa Recursos Genéticos e Biotecnologia. |
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Data corrente: |
12/12/2018 |
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Data da última atualização: |
02/07/2024 |
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Tipo da produção científica: |
Artigo em Periódico Indexado |
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Autoria: |
WOHLRES-VIANA, S.; ARASHIRO, E. K. N.; MINARE, T. P.; FERNANDES, C. A. C.; GRAZIA, J. G. V.; SIQUEIRA, L. G. B.; MACHADO, M. A.; VIANA, J. H. M. |
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Afiliação: |
UNIVERSIDADE FEDERAL DE JUIZ DE FORA; UNIVERSIDADE FEDERAL FLUMINENSE; UNIVERSIDADE JOSÉ DO ROSARIO VELLANO; UNIVERSIDADE JOSE DO ROSARIO VELLANO; UNIVERSIDADE FEDERAL DE MINAS GERAIS; LUIZ GUSTAVO BRUNO SIQUEIRA, CNPGL; MARCO ANTONIO MACHADO, CNPGL; JOAO HENRIQUE MOREIRA VIANA, CENARGEN. |
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Título: |
Differential expression of LHCGR and its isoforms is associated to the variability in superovulation responses of Gir cattle. |
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Ano de publicação: |
2019 |
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Fonte/Imprenta: |
Theriogenology, v. 126, p. 68-74, 2019. |
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DOI: |
10.1016/j.theriogenology.2018.12.004 |
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Idioma: |
Inglês |
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Conteúdo: |
Abstract The aim of this study was to evaluate the pattern of expression of LHCGR isoforms in Gir heifers characterized as good (10.3 ± 1.2 ova/embryos per flush, n = 5) or poor responders (1.1 ± 0.3 ova/embryos per flush, n = 5) to superovulation protocols. In both groups, an adapted ultrasound-guided follicular aspiration system was used to collect granulosa cells from 8 mm follicles formed either during a synchronized, non-stimulated follicular wave (no stimulation control, NS) or on the fourth day of a superovulation protocol (SOV) induced with 200 IU of pFSH. The recovered follicular fluid was centrifuged and granulosa cells were washed with NaCl 0.9% and kept in RNAlater®. RNA extraction was performed using a commercial RNeasy Micro Kit and eluted samples were quantified and reverse transcribed using the commercial Superscript III kit. cDNA samples were amplified by real-time PCR using a primer to target LH/hCG receptor gene - not selective for LHCGR isoforms (total LHCGR) - and four sets of isoforms selective primers (S1, S10, S10 + 11, and S11). Analyses were performed using the REST software and expression levels are shown as mean ± SEM. Under physiological conditions (NS), poor responders had a higher expression of total LHCGR (4.9 ± 1.7 fold-change, P < 0.01) as well as isoforms S10, S11 and S10 + 11, compared to good responders. In both phenotypes, superovulation down-regulated total LHCGR expression (-0.5 ± 0.2 and -0.9 ± 0.0 for good and poor responders, respectively; P < 0.05). However, in poor responders the exogenous FSH treatment up-regulated the S10 (2.4 ± 2.0; P < 0.05), S10 + 11 (3.8 ± 3.2; P < 0.01), and S1 isoforms (1.8 ± 1.3; P < 0.05), compared to good responders We conclude that down-regulation of total LHCGR, associated to up-regulation of their inactive isoforms, may have compromised follicle development and thus contributed to the low efficiency of superovulation in heifers with a poor responder phenotype. MenosAbstract The aim of this study was to evaluate the pattern of expression of LHCGR isoforms in Gir heifers characterized as good (10.3 ± 1.2 ova/embryos per flush, n = 5) or poor responders (1.1 ± 0.3 ova/embryos per flush, n = 5) to superovulation protocols. In both groups, an adapted ultrasound-guided follicular aspiration system was used to collect granulosa cells from 8 mm follicles formed either during a synchronized, non-stimulated follicular wave (no stimulation control, NS) or on the fourth day of a superovulation protocol (SOV) induced with 200 IU of pFSH. The recovered follicular fluid was centrifuged and granulosa cells were washed with NaCl 0.9% and kept in RNAlater®. RNA extraction was performed using a commercial RNeasy Micro Kit and eluted samples were quantified and reverse transcribed using the commercial Superscript III kit. cDNA samples were amplified by real-time PCR using a primer to target LH/hCG receptor gene - not selective for LHCGR isoforms (total LHCGR) - and four sets of isoforms selective primers (S1, S10, S10 + 11, and S11). Analyses were performed using the REST software and expression levels are shown as mean ± SEM. Under physiological conditions (NS), poor responders had a higher expression of total LHCGR (4.9 ± 1.7 fold-change, P < 0.01) as well as isoforms S10, S11 and S10 + 11, compared to good responders. In both phenotypes, superovulation down-regulated total LHCGR expression (-0.5 ± 0... Mostrar Tudo |
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Palavras-Chave: |
In vivo embryo production. |
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Thesaurus Nal: |
Embryo transfer; Luteinizing hormone. |
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Categoria do assunto: |
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L Ciência Animal e Produtos de Origem Animal |
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Marc: |
null
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Registro original: |
Embrapa Gado de Leite (CNPGL) |
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