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 | Acesso ao texto completo restrito à biblioteca da Embrapa Gado de Leite. Para informações adicionais entre em contato com rosangela.lacerda@embrapa.br. |
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Registro Completo |
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Biblioteca(s): |
Embrapa Gado de Leite. |
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Data corrente: |
17/01/2018 |
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Data da última atualização: |
27/01/2023 |
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Tipo da produção científica: |
Artigo em Periódico Indexado |
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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. |
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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. |
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Título: |
Sperm pretreatment with heparin and L-glutathione, sex-sorting, and double cryopreservation to improve intracytoplasmic sperm injection in bovine. |
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Ano de publicação: |
2017 |
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Fonte/Imprenta: |
Theriogenology, v. 93, p. 62-70, 2017. |
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Idioma: |
Inglês |
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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 |
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Palavras-Chave: |
ICSI; Pcx-EGFP; Sex-sorted sperm; Sperm pretreatment. |
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Thesaurus Nal: |
cattle. |
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Categoria do assunto: |
L Ciência Animal e Produtos de Origem Animal |
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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. |  | MACEDO, J. Os solos da regiao dos cerrados. In: ALVAREZ V., V.H.; FONTES, L.E.F.; FONTES, M.P.F. (Ed.). O solo nos grandes dominios morfoclimaticos do Brasil e o desenvolvimento sustentado. Vicosa: SBCS/UFV, 1996. p.135-155.| Biblioteca(s): Embrapa Cerrados. |
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| 4. | | MACEDO, J. The Brazilian cerrados. In: INTERNATIONAL WORKSHOP ON SOIL, WATER, AND NUTRIENT MANAGEMENT RESEARCH: environmental and productivity dimensions, 1994, Zschortau, Germany. Proceedings: the Zschortau plan for the implementation of soil, water, and nutrient ... . Zchortau: DSE/Bangkok: IBSRAM, 1995. p.78-86.| Biblioteca(s): Embrapa Cerrados. |
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| 5. | | MACEDO, J. Produção de alimentos: o potencial dos cerrados. Planaltina, DF: EMBRAPA-CPAC, 1996. 33 p. (EMBRAPA-CPAC. Documentos, 59). Apresentado no "Workshop de Ecologia e Biodiversidade", com o titulo "Prospectives for the rational use of the Brazilian's Cerrados for food production", Rio de Janeiro, 1994 (CRI4028). Ver CRI3752.| Biblioteca(s): Embrapa Cerrados. |
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| 13. | | MACEDO, J. R. de. Erosão em pastagens. Portal Dia de Campo, 18 ago. 2010. Disponível em : < http://www.diadecampo.com.br>. Acesso em: 09 fev. 2011.| Tipo: Artigo de Divulgação na Mídia |
| Biblioteca(s): Embrapa Solos. |
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| 16. | | MACEDO, J. L. V. de. Atividades de manejo na área experimental. In: SHIFT PROJECT ENV 23 (Manaus-AM). Recuperação de áreas degradadas e abandonadas, através de sistemas de policultivo. Manaus: EMBRAPA-CPAA: University of Hamburg, 1998. p. 1-2.| Tipo: Capítulo em Livro Técnico-Científico |
| Biblioteca(s): Embrapa Amazônia Ocidental. |
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| 19. | | MACEDO, J. L. V. de. Manejo da área experimental. In: SHIFT PROJECT ENV 23 (Manaus-AM). Recuperação de áreas degradadas e abandonadas, através de sistemas de policultivo. Manaus: Embrapa Amazônia Ocidental, 1999. p. 1-2. Relatorio Anual 1998.| Tipo: Capítulo em Livro Técnico-Científico |
| Biblioteca(s): Embrapa Amazônia Ocidental. |
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| Registros recuperados : 495 | |
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