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Vertebrate reproductive science and technology
RESEARCH ARTICLE

Effects of treatment with a microRNA mimic or inhibitor on the developmental competence, quality, epigenetic status and gene expression of buffalo (Bubalus bubalis) somatic cell nuclear transfer embryos

S. Sah A , A. K. Sharma A , S. K. Singla A , M. K. Singh https://orcid.org/0000-0003-4644-4443 A , M. S. Chauhan A , R. S. Manik A and P. Palta https://orcid.org/0000-0001-7021-8660 A B
+ Author Affiliations
- Author Affiliations

A Embryo Biotechnology Laboratory, Animal Biotechnology Centre, Indian Council of Agricultural Research–National Dairy Research Institute, GT Road, Karnal, Haryana, 132001 India.

B Corresponding author. Email: prabhatpalta@yahoo.com

Reproduction, Fertility and Development 32(5) 508-521 https://doi.org/10.1071/RD19084
Submitted: 1 March 2019  Accepted: 17 August 2019   Published: 21 January 2020

Abstract

Expression levels of 13 microRNAs (miRNAs) were compared between buffalo blastocysts produced by somatic cell nuclear transfer through hand-made cloning and IVF to improve cloning efficiency. Expression of miR-22, miR-145, miR-374a and miR-30c was higher, whereas that of miR-29b, miR-101, miR-302b, miR-34a, miR-21 and miR-25 was lower, in nuclear transferred (NT) than IVF embryos; the expression of miR-200b, miR-26a and miR-128 was similar between the two groups. Based on these, miR-145, which is involved in the regulation of pluripotency, was selected for further investigation of NT embryos. miR-145 expression was lowest at the 2-cell stage, increased through the 4-cell stage and was highest at the 8-cell or morula stage in a pattern that was similar between NT and IVF embryos. miR-145 expression was higher in NT than IVF embryos at all stages examined. Treatment of reconstructed embryos 1 h after electrofusion with an inhibitor of miR-145 for 1 h decreased the apoptotic index and increased the blastocyst rate, total cell number, ratio of cells in the inner cell mass to trophectoderm, global levels of acetylation of histone 3 at lysine 18 and expression of Krueppel-like factor 4 (KLF4), octamer-binding transcription factor 4 (OCT4) and SRY (sex determining region Y)-box 2 (SOX2) in blastocysts. Treatment with an miR-145 mimic had the opposite effects. In conclusion, treatment of NT embryos with an miR-145 inhibitor improves the developmental competence and quality, and increases histone acetylation and expression of pluripotency-related genes.

Graphical Abstract Image

Additional keywords: apoptosis, cloning, hand-made cloning, IVF.


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