Oxamflatin and ascorbic acid improves developmental competence and quality of buffalo (Bubalis bubalis) cloned embryos
Buffalo cloning is a powerful assisted reproductive tool for multiplying elite buffalo germplasm. However, the live off-spring production efficiency is low due to aberrant epigenetic reprogramming. Aberrant epigenetic marks can be modified by culturing donor cells and/or one cell stage fused embryo...
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Universidad del Zulia
2023-11-01
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| author | Sonal Gupta Gaurav Tripathi Kartikey Patel Ram Parsad Tanya Gupta Naresh L. Selokar Manoj Kumar Singh |
| author_facet | Sonal Gupta Gaurav Tripathi Kartikey Patel Ram Parsad Tanya Gupta Naresh L. Selokar Manoj Kumar Singh |
| author_sort | Sonal Gupta |
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Buffalo cloning is a powerful assisted reproductive tool for multiplying elite buffalo germplasm. However, the live off-spring production efficiency is low due to aberrant epigenetic reprogramming. Aberrant epigenetic marks can be modified by culturing donor cells and/or one cell stage fused embryo or both with epigenetic modifiers alone or in combination. In the present study, we examined the effect of oxamflatin (OxF), ascorbic acid (AA), and their combined (OxF+AA) effect on in vitro developmental competence, quality, and pregnancy establishment rate of buffalo cloned embryos. Oxamflatin is a histone deacetylase inhibitor, whereas ascorbic acid is a hypomethylating agent. To achieve this aim of the study, reconstructs (fused two enucleated ooplasm + donor cell) were cultured for 8 h, i.e., 4 h post-fusion and 4 h post-activation with 1 μM oxamflatin (OxF), 50 μM ascorbic acid (AA), and there combined (OxF+AA) treatment. There was no significant (p<0.05) difference in cleavage rates when reconstructs were treated with oxamflatin (81.34±0.81%), ascorbic acid (82.76±0.51%), combined treatment (82.17±0.54%) compared with control (82.87±0.63%). The blastocyst production rate was significantly higher (p<0.05) in combined treatment OxF+AA (41.64±0.95%) as compared to OxF (34.88±1.22%), AA (38.99±0.69%) and control (30.29±0.77%). The TUNEL assay showed a significantly lower (p<0.05) apoptotic index in combined (OxF+AA) treatment (1.43±0.43) as compared to oxamflatin (3.54±0.46), ascorbic acid (3.24±0.49) and control (5.06±0.48). The cloned embryos were transferred to the synchronized recipient (n=15 to18 buffaloes in each group), and the conception rate was observed better in combined treatment (OxF+AA) (46.66%) than oxamflatin (16.66%) and ascorbic acid (12.50%). At the same time, no pregnancy was reported in the control group. In conclusion, the combined treatment with oxamflatin and ascorbic acid improves the in vitro and in vivo developmental potential in buffalo-cloned embryos, which could probably be due to decreased methylation and increased acetylation of the embryos.
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| format | Article |
| id | doaj-art-00a471bbea92448d90cde5cbcde971b3 |
| institution | Kabale University |
| issn | 0798-2259 2521-9715 |
| language | English |
| publishDate | 2023-11-01 |
| publisher | Universidad del Zulia |
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| spelling | doaj-art-00a471bbea92448d90cde5cbcde971b32025-02-07T15:37:12ZengUniversidad del ZuliaRevista Científica0798-22592521-97152023-11-0133Suplemento10.52973/rcfcv-wbc128Oxamflatin and ascorbic acid improves developmental competence and quality of buffalo (Bubalis bubalis) cloned embryosSonal Gupta0Gaurav Tripathi1Kartikey Patel2Ram Parsad3Tanya Gupta4Naresh L. Selokar5Manoj Kumar Singh6Embryo Biotechnology Lab, Animal Biotechnology Division ICAR- National Dairy Research Institute, Karnal-132001, IndiaEmbryo Biotechnology Lab, Animal Biotechnology Division ICAR- National Dairy Research Institute, Karnal-132001, IndiaEmbryo Biotechnology Lab, Animal Biotechnology Division ICAR- National Dairy Research Institute, Karnal-132001, IndiaEmbryo Biotechnology Lab, Animal Biotechnology Division ICAR- National Dairy Research Institute, Karnal-132001, IndiaEmbryo Biotechnology Lab, Animal Biotechnology Division ICAR- National Dairy Research Institute, Karnal-132001, IndiaEmbryo Biotechnology Lab, Animal Biotechnology Division ICAR- National Dairy Research Institute, Karnal-132001, IndiaEmbryo Biotechnology Lab, Animal Biotechnology Division ICAR- National Dairy Research Institute, Karnal-132001, India Buffalo cloning is a powerful assisted reproductive tool for multiplying elite buffalo germplasm. However, the live off-spring production efficiency is low due to aberrant epigenetic reprogramming. Aberrant epigenetic marks can be modified by culturing donor cells and/or one cell stage fused embryo or both with epigenetic modifiers alone or in combination. In the present study, we examined the effect of oxamflatin (OxF), ascorbic acid (AA), and their combined (OxF+AA) effect on in vitro developmental competence, quality, and pregnancy establishment rate of buffalo cloned embryos. Oxamflatin is a histone deacetylase inhibitor, whereas ascorbic acid is a hypomethylating agent. To achieve this aim of the study, reconstructs (fused two enucleated ooplasm + donor cell) were cultured for 8 h, i.e., 4 h post-fusion and 4 h post-activation with 1 μM oxamflatin (OxF), 50 μM ascorbic acid (AA), and there combined (OxF+AA) treatment. There was no significant (p<0.05) difference in cleavage rates when reconstructs were treated with oxamflatin (81.34±0.81%), ascorbic acid (82.76±0.51%), combined treatment (82.17±0.54%) compared with control (82.87±0.63%). The blastocyst production rate was significantly higher (p<0.05) in combined treatment OxF+AA (41.64±0.95%) as compared to OxF (34.88±1.22%), AA (38.99±0.69%) and control (30.29±0.77%). The TUNEL assay showed a significantly lower (p<0.05) apoptotic index in combined (OxF+AA) treatment (1.43±0.43) as compared to oxamflatin (3.54±0.46), ascorbic acid (3.24±0.49) and control (5.06±0.48). The cloned embryos were transferred to the synchronized recipient (n=15 to18 buffaloes in each group), and the conception rate was observed better in combined treatment (OxF+AA) (46.66%) than oxamflatin (16.66%) and ascorbic acid (12.50%). At the same time, no pregnancy was reported in the control group. In conclusion, the combined treatment with oxamflatin and ascorbic acid improves the in vitro and in vivo developmental potential in buffalo-cloned embryos, which could probably be due to decreased methylation and increased acetylation of the embryos. https://www.produccioncientificaluz.org/index.php/cientifica/article/view/43517oxamflatinascorbic acidbuffalo cloningepigenetic reprogrammingpregnancy |
| spellingShingle | Sonal Gupta Gaurav Tripathi Kartikey Patel Ram Parsad Tanya Gupta Naresh L. Selokar Manoj Kumar Singh Oxamflatin and ascorbic acid improves developmental competence and quality of buffalo (Bubalis bubalis) cloned embryos Revista Científica oxamflatin ascorbic acid buffalo cloning epigenetic reprogramming pregnancy |
| title | Oxamflatin and ascorbic acid improves developmental competence and quality of buffalo (Bubalis bubalis) cloned embryos |
| title_full | Oxamflatin and ascorbic acid improves developmental competence and quality of buffalo (Bubalis bubalis) cloned embryos |
| title_fullStr | Oxamflatin and ascorbic acid improves developmental competence and quality of buffalo (Bubalis bubalis) cloned embryos |
| title_full_unstemmed | Oxamflatin and ascorbic acid improves developmental competence and quality of buffalo (Bubalis bubalis) cloned embryos |
| title_short | Oxamflatin and ascorbic acid improves developmental competence and quality of buffalo (Bubalis bubalis) cloned embryos |
| title_sort | oxamflatin and ascorbic acid improves developmental competence and quality of buffalo bubalis bubalis cloned embryos |
| topic | oxamflatin ascorbic acid buffalo cloning epigenetic reprogramming pregnancy |
| url | https://www.produccioncientificaluz.org/index.php/cientifica/article/view/43517 |
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