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Buffalo (Bubalus bubalis) SCNT embryos produced from somatic cells isolated from frozen–thawed semen: effect of trichostatin A on the in vitro and in vivo developmental potential, quality and epigenetic status

Published online by Cambridge University Press:  27 October 2015

Naresh L. Selokar ,
Monika Saini ,
Himanshu Agrawal ,
Prabhat Palta ,
Manmohan S. Chauhan ,
Radheysham Manik  and
Suresh K Singla
Naresh L. Selokar
Affiliation:
Animal Biotechnology Centre, National Dairy Research Institute, Karnal 132001, India. Department of Animal Physiology and Reproduction, Central Institute for Research on Buffaloes, Hisar 125011, India.
Monika Saini
Affiliation:
Animal Biotechnology Centre, National Dairy Research Institute, Karnal 132001, India. Department of Animal Physiology and Reproduction, Central Institute for Research on Buffaloes, Hisar 125011, India.
Himanshu Agrawal
Affiliation:
Animal Biotechnology Centre, National Dairy Research Institute, Karnal 132001, India.
Prabhat Palta
Affiliation:
Animal Biotechnology Centre, National Dairy Research Institute, Karnal 132001, India.
Manmohan S. Chauhan
Affiliation:
Animal Biotechnology Centre, National Dairy Research Institute, Karnal 132001, India.
Radheysham Manik
Affiliation:
Animal Biotechnology Centre, National Dairy Research Institute, Karnal 132001, India.
Suresh K Singla*
Affiliation:
Animal Biotechnology Centre, National Dairy Research Institute, Karnal, India.
*
All correspondence to: S.K. Singla. Animal Biotechnology Centre, National Dairy Research Institute, Karnal, India. E-mail: singlasuresh@yahoo.com
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Summary

This study examined the effects of trichostatin A (TSA) treatment of reconstructed buffalo embryos, produced by hand-made cloning using somatic cells isolated from over a decade old frozen–thawed semen, on their in vitro and in vivo developmental competence, quality and epigenetic status. Following treatment of reconstructed embryos with TSA (0, 50 or 75 nM) for 10 h prior to culture, the cleavage (100.0 ± 0, 94.5 ± 2.3 and 96.1 ± 1.2%, respectively) and blastocyst rate (50.6 ± 2.3, 48.4 ± 2.7 and 48.1 ± 2.6%, respectively), total cell number (275 ± 17.4, 289 ± 30.1 and 317 ± 24.2, respectively) and apoptotic index (5.6 ± 0.7, 3.4 ± 0.9 and 4.5 ± 1.4, respectively) were not significantly different among the three groups. However, TSA treatment increased (P < 0.05) the global level of H4K5ac and decreased (P < 0.05) that of H3K27me3 in blastocysts whereas the global level of H3K18ac was not affected significantly. Transfer of embryos treated with 75 nM TSA (n = 10) to recipients resulted in two pregnancies (20%), one out of which was aborted in the second and the other in the third trimester whereas transfer of control embryos (n = 20) or those treated with 50 nM TSA (n = 12) did not result in any pregnancy. In conclusion, these results suggest that TSA treatment of cloned buffalo embryos produced using somatic cells isolated from frozen–thawed semen improved their epigenetic status but not the in vitro developmental potential and offspring rate.

Keywords

BuffaloEmbryoHand-made cloningSCNTTrichostatin A

Information

Type
Short Communication
Information
Zygote , Volume 24 , Issue 4 , August 2016 , pp. 549 - 553
Copyright
Copyright © Cambridge University Press 2015 

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