Hostname: page-component-78c5997874-lj6df Total loading time: 0 Render date: 2024-11-11T09:01:44.172Z Has data issue: false hasContentIssue false
  • English
  • Français

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
Get access Rights & Permissions [Opens in a new window]

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

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Campbell, K.H., Fisher, P., Chen, W.C., Choi, I., Kelly, R.D., Lee, J.H. & Xhu, J. (2007). Somatic cell nuclear transfer: past, present and future perspectives. Theriogenology 68, S214–31.Google Scholar
Cedar, H., & Bergman, Y. (2009). Linking DNA methylation and histone modification: patterns and paradigms. Nat. Rev. Genet. 10, 295–304.Google Scholar
Ding, X., Wang, Y., Zhang, D., Wang, Y., Guo, Z. & Zhang, Y. (2008). Increased preimplantation development of cloned bovine embryos treated with 5-aza-20-deoxycytidine and trichostatin A. Theriogenology 70, 622–30.Google Scholar
Enright, B.P., Kubota, C., Yang, X. & Tian, X.C. (2003). Epigenetic characteristics and development of embryos cloned from donor cells treated by trichostatin A or 5-aza-29-deoxycytidine. Biol. Reprod. 69, 896–901.Google Scholar
Iager, A.E., Ragina, N.P., Ross, P.J., Beyhan, Z., Cunniff, K., Rodriguez, R.M., & Cibelli, J.B. (2008). Trichostatin A improves histone acetylation in bovine somatic cell nuclear transfer early embryos. Cloning Stem Cells 10, 371–9.CrossRefGoogle ScholarPubMed
Luo, C., Lu, F., Wang, X., Wang, Z., Li, X., Gong, F., Jiang, J., Liu, Q., & Shi, D. (2013). Treatment of donor cells with trichostatin A improves in vitro development and reprogramming of buffalo (Bubalus bubalis) nucleus transfer embryos. Theriogenology 80, 878–86.CrossRefGoogle ScholarPubMed
Oback, B. & Wells, , D. (2002). Donor cells for nuclear cloning: many are called, but few are chosen. Cloning and Stem Cells 4, 147–68.Google Scholar
Saini, M., Selokar, N.L., Agrawal, H., Singla, S.K., Chauhan, M.S., Manik, R.S. & Palta, P. (2014). Treatment of buffalo (Bubalus bubalis) donor cells with trichostatin A and 5-aza-2. Reprod. Fertil. Dev. doi: 10.1071/RD14176. [Epub ahead of print]Google Scholar
Saini, M., Selokar, N.L., Raja, A.K., Sahare, A.A., Singla, S.K., Chauhan, M.S., Manik, R.S. & Palta, P. (2015). Effect of donor cell type on developmental competence, quality, gene expression, and epigenetic status of interspecies cloned embryos produced using cells from wild buffalo and oocytes from domestic buffalo. Theriogenology 84, 101–8e1.CrossRefGoogle ScholarPubMed
Selokar, N.L., Saini, M., Palta, P., Chauhan, M.S., Manik, R.S. & Singla, S.K. (2014). Hope for restoration of dead valuable bulls through cloning using donor somatic cells isolated from cryopreserved semen. PLoS One 9, e90755.Google Scholar
Srirattana, K., Ketudat-Cairns, M., Nagai, T., Kaneda, M. & Parnpai, R. (2014). Effects of trichostatin A on In vitro development and DNA methylation level of the satellite I region of swamp buffalo (Bubalus bubalis) cloned embryos. J. Reprod. Dev. 60, 336–41.Google Scholar
Tian, X.C., Kubota, C., Enright, B. & Yang, X. (2003). Cloning animals by somatic cell nuclear transfer—biological factors. Reprod. Biol. Endocrinol. 1, 98.CrossRefGoogle ScholarPubMed
Zhang, Y.H., Song, E.S., Kim, E.S., Cong, P.Q., Lee, S.H., Lee, J.W., Yi, Y.J. & Park, C.S. (2009). Effects of donor cell passage, size and type on development of porcine embryos derived from somatic cell nuclear transfer. Asian Austral J Anim 22, 194–200.CrossRefGoogle Scholar