J Vet Sci.  2015 Sep;16(3):373-380. 10.4142/jvs.2015.16.3.373.

Rapamycin treatment during in vitro maturation of oocytes improves embryonic development after parthenogenesis and somatic cell nuclear transfer in pigs

• Lee J
• Park JI
• Yun JI
• Lee Y
• Yong H
• Lee ST
• Park CK
• Hyun SH
• Lee GS
• Lee E

• KMID: 2344311
• DOI: http://doi.org/10.4142/jvs.2015.16.3.373

Abstract

This study was conducted to investigate the effects of rapamycin treatment during in vitro maturation (IVM) on oocyte maturation and embryonic development after parthenogenetic activation (PA) and somatic cell nuclear transfer (SCNT) in pigs. Morphologically good (MGCOCs) and poor oocytes (MPCOCs) were untreated or treated with 1 nM rapamycin during 0-22 h, 22-42 h, or 0-42 h of IVM. Rapamycin had no significant effects on nuclear maturation and blastocyst formation after PA of MGCOCs. Blastocyst formation after PA was significantly increased by rapamycin treatment during 22-42 h and 0-42 h (46.6% and 46.5%, respectively) relative to the control (33.3%) and 0-22 h groups (38.6%) in MPCOCs. In SCNT, blastocyst formation tended to increase in MPCOCs treated with rapamycin during 0-42 h of IVM relative to untreated oocytes (20.3% vs. 14.3%, 0.05 < p < 0.1), while no improvement was observed in MGCOCs. Gene expression analysis revealed that transcript abundance of Beclin 1 and microtubule-associated protein 1 light chain 3 mRNAs was significantly increased in MPCOCs by rapamycin relative to the control. Our results demonstrated that autophagy induction by rapamycin during IVM improved developmental competence of oocytes derived from MPCOCs.

Keyword

autophagy; oocyte maturation; pig; rapamycin; somatic cell nuclear transfer

MeSH Terms

Animals
Embryonic Development/*drug effects
Female
In Vitro Oocyte Maturation Techniques/veterinary
Nuclear Transfer Techniques/*veterinary
Oocytes/growth & development
*Parthenogenesis
Sirolimus/*pharmacology
Sus scrofa/*growth & development/metabolism
Sirolimus

Figure

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  Fig. 1 (A) Morphologically good (MGCOCs) and (B) poor pig oocytes (MPCOCs) used in this study. MGCOCs were surrounded by a thick cumulus cell layer, whereas MPCOCs were partially denuded or had a thin cumulus cell layer compared to MGCOCs. Scale bars = 100 µm.

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  Fig. 2 Relative transcript abundance (mean ± SEM) of Beclin 1 and LC3 mRNAs in metaphase II oocytes derived from morphologically good oocytes (MGCOCs) and morphologically poor oocytes (MPCOCs) treated with 1 nM rapamycin during in vitro maturation. Bars with different letters (a, b) in the same mRNA are significantly different (p < 0.05).

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