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Registro Completo |
Biblioteca(s): |
Embrapa Gado de Leite. |
Data corrente: |
17/01/2018 |
Data da última atualização: |
27/01/2023 |
Tipo da produção científica: |
Artigo em Periódico Indexado |
Autoria: |
CANEL, N. G.; BEVACQUA, R. J.; HIRIART, M. I.; RABELO, N. C.; CAMARGO, L. S. de A.; ROMANATO, M.; CALVO, L. P.; SALAMONE, D. F. |
Afiliação: |
Natália Gabriela Canel, Universidade de Buenos Aires; Romina Jimena Bevacqua, Universidade de Buenos Aires; María Inés Hiriart, Universidade de Buenos Aires; Natana Chaves Rabelo, UFJF; LUIZ SERGIO DE ALMEIDA CAMARGO, CNPGL; Marina Romanato, CONICET; Lucrecia Pineiro de Calvo, CONICET; Daniel Felipe Salamone, Universidade de Buenos Aires. |
Título: |
Sperm pretreatment with heparin and L-glutathione, sex-sorting, and double cryopreservation to improve intracytoplasmic sperm injection in bovine. |
Ano de publicação: |
2017 |
Fonte/Imprenta: |
Theriogenology, v. 93, p. 62-70, 2017. |
Idioma: |
Inglês |
Conteúdo: |
Abstract In bovine, intracytoplasmic sperm injection (ICSI) remains inefficient partially due to low levels of sperm decondensation. The aim of this study was to determine whether the injection of normal size sperm pretreated with heparin (Hep) and l-glutathione (GSH), the use of sex-sorted sperm, the double round of sperm freezing/thawing (re frozen), or the combination of these approaches can improve sperm decondensation and embryo development after ICSI in cattle. Cleavage and blastocyst rates were evaluated on days 2 and 7 post ICSI. Quality of ICSI blastocysts was analyzed by the relative expression of four genes by qPCR and the DNA fragmentation index by TUNEL assay. For all assays, semen samples were co-incubated with pCX-EGFP 50 ng/μl before ICSI. GFP expression, which can be detected by fluorescence microscopy, was used as a tool to estimate the success of sperm decondensation after ICSI. The use of normal size sperm pretreated with 80 μM Hep-15 mM GSH for 20 h (Hep-GSH) increased cleavage, blastocyst and EGFP + blastocysts rates (60.8, 19.4 and 61.9%) compared to control ICSI (35, 4.9 and 20%) (p < 0.05). Moreover, HMGN1, GLUT5, AQP3 and POU5F1 transcription levels did not differ between ICSI Hep-GSH and IVF embryos. The use of sex-sorted sperm (X, Y) improved cleavage rates and EGFP expression at day 4 (83 and 30.2% for ICSI Y and 83.2 and 31.7% for ICSI X) compared to non-sorted group (50.9 and 15.1%), not showing differences at the blastocyst stage. Finally, sex sorting (X) was combined with Hep-GSH and/or re frozen treatments. The use of Hep-GSH diminished cleavage rates from ICSI X re frozen group (80.4% vs. 94.2%) and blastocyst development from ICSI X group (3.3% vs. 10%), compared with their controls (p < 0.05). While Hep-GSH pretreatment induced lower transgene expression at day 4, no differences were found at the blastocyst stage between ICSI groups (from 58.3 to 80%). TUNEL assay showed higher DNA fragmentation indexes for all ICSI treatments (p < 0.05), except for ICSI X Hep-GSH, which did not differ from IVF X control. In conclusion, the use of normal size sperm pretreated with Hep-GSH, combined or not with sex-sorting by flow cytometry could improve ICSI outcomes in cattle. MenosAbstract In bovine, intracytoplasmic sperm injection (ICSI) remains inefficient partially due to low levels of sperm decondensation. The aim of this study was to determine whether the injection of normal size sperm pretreated with heparin (Hep) and l-glutathione (GSH), the use of sex-sorted sperm, the double round of sperm freezing/thawing (re frozen), or the combination of these approaches can improve sperm decondensation and embryo development after ICSI in cattle. Cleavage and blastocyst rates were evaluated on days 2 and 7 post ICSI. Quality of ICSI blastocysts was analyzed by the relative expression of four genes by qPCR and the DNA fragmentation index by TUNEL assay. For all assays, semen samples were co-incubated with pCX-EGFP 50 ng/μl before ICSI. GFP expression, which can be detected by fluorescence microscopy, was used as a tool to estimate the success of sperm decondensation after ICSI. The use of normal size sperm pretreated with 80 μM Hep-15 mM GSH for 20 h (Hep-GSH) increased cleavage, blastocyst and EGFP + blastocysts rates (60.8, 19.4 and 61.9%) compared to control ICSI (35, 4.9 and 20%) (p < 0.05). Moreover, HMGN1, GLUT5, AQP3 and POU5F1 transcription levels did not differ between ICSI Hep-GSH and IVF embryos. The use of sex-sorted sperm (X, Y) improved cleavage rates and EGFP expression at day 4 (83 and 30.2% for ICSI Y and 83.2 and 31.7% for ICSI X) compared to non-sorted group (50.9 and 15.1%), not showing differences at the blastocyst stage. Fi... Mostrar Tudo |
Palavras-Chave: |
ICSI; Pcx-EGFP; Sex-sorted sperm; Sperm pretreatment. |
Thesaurus Nal: |
cattle. |
Categoria do assunto: |
L Ciência Animal e Produtos de Origem Animal |
Marc: |
LEADER 03038naa a2200265 a 4500 001 2085654 005 2023-01-27 008 2017 bl uuuu u00u1 u #d 100 1 $aCANEL, N. G. 245 $aSperm pretreatment with heparin and L-glutathione, sex-sorting, and double cryopreservation to improve intracytoplasmic sperm injection in bovine.$h[electronic resource] 260 $c2017 520 $aAbstract In bovine, intracytoplasmic sperm injection (ICSI) remains inefficient partially due to low levels of sperm decondensation. The aim of this study was to determine whether the injection of normal size sperm pretreated with heparin (Hep) and l-glutathione (GSH), the use of sex-sorted sperm, the double round of sperm freezing/thawing (re frozen), or the combination of these approaches can improve sperm decondensation and embryo development after ICSI in cattle. Cleavage and blastocyst rates were evaluated on days 2 and 7 post ICSI. Quality of ICSI blastocysts was analyzed by the relative expression of four genes by qPCR and the DNA fragmentation index by TUNEL assay. For all assays, semen samples were co-incubated with pCX-EGFP 50 ng/μl before ICSI. GFP expression, which can be detected by fluorescence microscopy, was used as a tool to estimate the success of sperm decondensation after ICSI. The use of normal size sperm pretreated with 80 μM Hep-15 mM GSH for 20 h (Hep-GSH) increased cleavage, blastocyst and EGFP + blastocysts rates (60.8, 19.4 and 61.9%) compared to control ICSI (35, 4.9 and 20%) (p < 0.05). Moreover, HMGN1, GLUT5, AQP3 and POU5F1 transcription levels did not differ between ICSI Hep-GSH and IVF embryos. The use of sex-sorted sperm (X, Y) improved cleavage rates and EGFP expression at day 4 (83 and 30.2% for ICSI Y and 83.2 and 31.7% for ICSI X) compared to non-sorted group (50.9 and 15.1%), not showing differences at the blastocyst stage. Finally, sex sorting (X) was combined with Hep-GSH and/or re frozen treatments. The use of Hep-GSH diminished cleavage rates from ICSI X re frozen group (80.4% vs. 94.2%) and blastocyst development from ICSI X group (3.3% vs. 10%), compared with their controls (p < 0.05). While Hep-GSH pretreatment induced lower transgene expression at day 4, no differences were found at the blastocyst stage between ICSI groups (from 58.3 to 80%). TUNEL assay showed higher DNA fragmentation indexes for all ICSI treatments (p < 0.05), except for ICSI X Hep-GSH, which did not differ from IVF X control. In conclusion, the use of normal size sperm pretreated with Hep-GSH, combined or not with sex-sorting by flow cytometry could improve ICSI outcomes in cattle. 650 $acattle 653 $aICSI 653 $aPcx-EGFP 653 $aSex-sorted sperm 653 $aSperm pretreatment 700 1 $aBEVACQUA, R. J. 700 1 $aHIRIART, M. I. 700 1 $aRABELO, N. C. 700 1 $aCAMARGO, L. S. de A. 700 1 $aROMANATO, M. 700 1 $aCALVO, L. P. 700 1 $aSALAMONE, D. F 773 $tTheriogenology$gv. 93, p. 62-70, 2017.
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Embrapa Gado de Leite (CNPGL) |
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1. | | SANTOS, L. P.; GARCIA, M. V. B.; GARCIA, T. B.; BARRONCAS, M. J. K.; GUEDES, F. M. de S.; VIEIRA, V. V. Avaliação do desempenho de cultivares de mamoeiros, introduzidas do Estado do Espírito Santo, em condições edafoclimáticas do Amazonas. In: REUNIÃO TÉCNICA DE PESQUISA E DESENVOLVIMENTO DA EMBRAPA AMAZÔNIA OCIDENTAL, 4., 2009, Manaus. Anais... Manaus: Embrapa Amazônia Ocidental, 2009. p. 96-98. (Embrapa Amazônia Ocidental. Documentos, 73).Tipo: Artigo em Anais de Congresso / Nota Técnica |
Biblioteca(s): Embrapa Amazônia Ocidental. |
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2. | | SANTOS, L. P.; GARCIA, M. V. B.; GARCIA, T. B.; BARRONCAS, M. J. K.; GUEDES, F. M. de S.; VIEIRA, V. V. Análises de crescimento e produtividade do mamoeiro cultivado com uso de irrigação por gotejamento superficial no Estado do Amazonas. In: REUNIÃO TÉCNICA DE PESQUISA E DESENVOLVIMENTO DA EMBRAPA AMAZÔNIA OCIDENTAL, 4., 2009, Manaus. Anais... Manaus: Embrapa Amazônia Ocidental, 2009. p. 99-102. (Embrapa Amazônia Ocidental. Documentos, 73).Tipo: Artigo em Anais de Congresso / Nota Técnica |
Biblioteca(s): Embrapa Amazônia Ocidental. |
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3. | | SANTOS, L. P.; SILVA, E. de B.; VIEIRA, V. V.; GUEDES, F. M. DE S.; GARCIA, T. B.; GARCIA, M. V. B.; CATTANEO, L. F.; FERREGUETTI, G. A.; BRAGANÇA, S. M. Comportamento diferencial do estado nutricional de cultivares de mamoeiros avaliados por meio das análises de macronutrientes em limbos e pecíolos foliares em condições edafoclimáticas do Amazonas. In: CONGRESSO BRASILEIRO DE FRUTICULTURA, 21., 2010, Natal. Frutas: saúde, inovação e responsabilidade: anais. Natal: Sociedade Brasileira de Fruticultura, 2010.Tipo: Artigo em Anais de Congresso |
Biblioteca(s): Embrapa Amazônia Ocidental. |
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4. | | SANTOS, L. P.; SILVA, E. de B.; VIEIRA, V. V.; GUEDES, F. M. DE S.; GARCIA, T. B.; GARCIA, M. V. B.; CATTANEO, L. F.; FERREGUETTI, G. A.; BRAGANÇA, S. M. Comportamento diferencial do estado nutricional de cultivares de mamoeiros avaliados por meio das análises de micronutrientes em limbos e pecíolos foliares em condições edafoclimáticas do Amazonas. In: CONGRESSO BRASILEIRO DE FRUTICULTURA, 21., 2010, Natal. Frutas: saúde, inovação e responsabilidade: anais. Natal: Sociedade Brasileira de Fruticultura, 2010.Tipo: Artigo em Anais de Congresso |
Biblioteca(s): Embrapa Amazônia Ocidental. |
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