The effect of different zwitterionic buffers and PBS used for out-of-incubator procedures during standard in vitro embryo production on development, morphology and gene expression of bovine embryos
The effect of the zwitterionic buffers HEPES, TES and MOPS and of PBS used for out-of-incubator procedures during standard in vitro embryo production on bovine oocytes and embryo development, morphology and on the expression patterns of eight selected genes Fgf-4, Lama1, Ube2a, Gsta4, Il6, Sod1, Prss11 and Hspb1, was evaluated. All buffers were prepared at a concentration of 10 mM in TALP medium, with the exception of PBS. The total time of oocyte/embryo exposure to each buffer was ∼41 min. The cleavage rates and number of embryos that developed to ≥8 cells at day 4 were no different among the buffers tested, however, more blastocysts developed at day 7, 8 and 9 in HEPES and MOPS treatments than in PBS and TES (P < 0.05). No difference between buffers in total and apoptotic cell number was found. Except for Hspb1 and Ube2a genes, the levels of expression of the six remaining transcripts were higher in in vivo than in in vitro embryos irrespective of buffer used (P < 0.05). In addition, higher expression of Hspb1 and lower expression of Ube2a and Lama1 were observed in PBS and TES than in MOPS and HEPES treatments (P < 0.05). Expression of Fgf-4 and Gsta4 in the in vitro embryos was lower in PBS than in the remaining three buffers (P < 0.05) and the level of expression of the Il6 gene was not affected by any buffer tested but was lower in in vitro than in in vivo derived embryos. Expression of both Sod1 and Prss11 genes in MOPS were at the level of the in vivo embryos. These results showed that the choice of buffer and short exposure time of ∼41 min, affects mRNA expression of in vitro produced bovine embryos. © 2008 Elsevier Inc. All rights reserved.
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| Subjects: | Buffers, Zwitteronic, Embryo, Culture, Apoptosis, Gene expression, |
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dig-inia-es-10261-2929032023-02-20T07:33:37Z The effect of different zwitterionic buffers and PBS used for out-of-incubator procedures during standard in vitro embryo production on development, morphology and gene expression of bovine embryos Palasz, A. T. Breña, P. B. De la Fuente, J. Gutiérrez Adán, Alfonso Buffers Zwitteronic Embryo Culture Apoptosis Gene expression The effect of the zwitterionic buffers HEPES, TES and MOPS and of PBS used for out-of-incubator procedures during standard in vitro embryo production on bovine oocytes and embryo development, morphology and on the expression patterns of eight selected genes Fgf-4, Lama1, Ube2a, Gsta4, Il6, Sod1, Prss11 and Hspb1, was evaluated. All buffers were prepared at a concentration of 10 mM in TALP medium, with the exception of PBS. The total time of oocyte/embryo exposure to each buffer was ∼41 min. The cleavage rates and number of embryos that developed to ≥8 cells at day 4 were no different among the buffers tested, however, more blastocysts developed at day 7, 8 and 9 in HEPES and MOPS treatments than in PBS and TES (P < 0.05). No difference between buffers in total and apoptotic cell number was found. Except for Hspb1 and Ube2a genes, the levels of expression of the six remaining transcripts were higher in in vivo than in in vitro embryos irrespective of buffer used (P < 0.05). In addition, higher expression of Hspb1 and lower expression of Ube2a and Lama1 were observed in PBS and TES than in MOPS and HEPES treatments (P < 0.05). Expression of Fgf-4 and Gsta4 in the in vitro embryos was lower in PBS than in the remaining three buffers (P < 0.05) and the level of expression of the Il6 gene was not affected by any buffer tested but was lower in in vitro than in in vivo derived embryos. Expression of both Sod1 and Prss11 genes in MOPS were at the level of the in vivo embryos. These results showed that the choice of buffer and short exposure time of ∼41 min, affects mRNA expression of in vitro produced bovine embryos. © 2008 Elsevier Inc. All rights reserved. 2023-02-20T07:33:37Z 2023-02-20T07:33:37Z 2008 artículo Theriogenology 70(9): 1461-1470 (2008) 0093-691X http://hdl.handle.net/20.500.12792/3291 http://hdl.handle.net/10261/292903 10.1016/j.theriogenology.2008.06.092 1879-3231 en none Elsevier |
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Buffers Zwitteronic Embryo Culture Apoptosis Gene expression Buffers Zwitteronic Embryo Culture Apoptosis Gene expression |
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Buffers Zwitteronic Embryo Culture Apoptosis Gene expression Buffers Zwitteronic Embryo Culture Apoptosis Gene expression Palasz, A. T. Breña, P. B. De la Fuente, J. Gutiérrez Adán, Alfonso The effect of different zwitterionic buffers and PBS used for out-of-incubator procedures during standard in vitro embryo production on development, morphology and gene expression of bovine embryos |
| description |
The effect of the zwitterionic buffers HEPES, TES and MOPS and of PBS used for out-of-incubator procedures during standard in vitro embryo production on bovine oocytes and embryo development, morphology and on the expression patterns of eight selected genes Fgf-4, Lama1, Ube2a, Gsta4, Il6, Sod1, Prss11 and Hspb1, was evaluated. All buffers were prepared at a concentration of 10 mM in TALP medium, with the exception of PBS. The total time of oocyte/embryo exposure to each buffer was ∼41 min. The cleavage rates and number of embryos that developed to ≥8 cells at day 4 were no different among the buffers tested, however, more blastocysts developed at day 7, 8 and 9 in HEPES and MOPS treatments than in PBS and TES (P < 0.05). No difference between buffers in total and apoptotic cell number was found. Except for Hspb1 and Ube2a genes, the levels of expression of the six remaining transcripts were higher in in vivo than in in vitro embryos irrespective of buffer used (P < 0.05). In addition, higher expression of Hspb1 and lower expression of Ube2a and Lama1 were observed in PBS and TES than in MOPS and HEPES treatments (P < 0.05). Expression of Fgf-4 and Gsta4 in the in vitro embryos was lower in PBS than in the remaining three buffers (P < 0.05) and the level of expression of the Il6 gene was not affected by any buffer tested but was lower in in vitro than in in vivo derived embryos. Expression of both Sod1 and Prss11 genes in MOPS were at the level of the in vivo embryos. These results showed that the choice of buffer and short exposure time of ∼41 min, affects mRNA expression of in vitro produced bovine embryos. © 2008 Elsevier Inc. All rights reserved. |
| format |
artículo |
| topic_facet |
Buffers Zwitteronic Embryo Culture Apoptosis Gene expression |
| author |
Palasz, A. T. Breña, P. B. De la Fuente, J. Gutiérrez Adán, Alfonso |
| author_facet |
Palasz, A. T. Breña, P. B. De la Fuente, J. Gutiérrez Adán, Alfonso |
| author_sort |
Palasz, A. T. |
| title |
The effect of different zwitterionic buffers and PBS used for out-of-incubator procedures during standard in vitro embryo production on development, morphology and gene expression of bovine embryos |
| title_short |
The effect of different zwitterionic buffers and PBS used for out-of-incubator procedures during standard in vitro embryo production on development, morphology and gene expression of bovine embryos |
| title_full |
The effect of different zwitterionic buffers and PBS used for out-of-incubator procedures during standard in vitro embryo production on development, morphology and gene expression of bovine embryos |
| title_fullStr |
The effect of different zwitterionic buffers and PBS used for out-of-incubator procedures during standard in vitro embryo production on development, morphology and gene expression of bovine embryos |
| title_full_unstemmed |
The effect of different zwitterionic buffers and PBS used for out-of-incubator procedures during standard in vitro embryo production on development, morphology and gene expression of bovine embryos |
| title_sort |
effect of different zwitterionic buffers and pbs used for out-of-incubator procedures during standard in vitro embryo production on development, morphology and gene expression of bovine embryos |
| publisher |
Elsevier |
| publishDate |
2008 |
| url |
http://hdl.handle.net/20.500.12792/3291 http://hdl.handle.net/10261/292903 |
| work_keys_str_mv |
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