甲基化酶抑制剂5-Aza-CdR处理对德保猪手工克隆胚胎体外发育潜能的影响

doi:10.11843/j.issn.0366-6964.2014.08.010

Abstract

Abstract:

The inefficiency of cloned animal producing is mainly due to uncompleted epigenetic reprogramming. In this study, the effect of Debao pig ear fibroblasts treated with different concentrations (0.00（control）, 0.25, 0.50 and 1.00 μmol•L^-1) of 5-Aza-CdR (DNA methyltransferase inhibitor) on the developmental potency of handmade cloned pig embryos were investigated. The results showed that all treated cells have “S” shape growth curve, meantime cells treated with 0.25 μmol•L-1 5-Aza-CdR have a growth curve which was similar to that of control group’s. With the concentrations increasing of 5-Aza-CdR, cells showed distortion, growth inhibition in a certain degree, and even death. Debao pig fibroblasts treated with 5-Aza-CdR from 0.00 to 0.50 μmol•L^-1 didn’t show significantly alteration on the chromosome ploidy.5-Aza-CdR could down regulate the expression of Dnmt1, Tet1, Tet2 and Tet3 on Debao pig ear fibroblasts following a dose-related quantity pattern. The cleavage and morula rates of handmade cloned embryos with donor cell treated with 5-Aza-CdR had no significant difference with that of the control group (P>0.05). However, the blastocyst rates of 0.25 μmol•L^-1 and 0.50 μmol•L^-1 treatment groups were significantly higher than that of 1.00 μmol•L^-1 group (P<0.05), but had no significant difference with that of the control group (P>0.05). The total blastocyst cells number of 0.50 μmol•L^-1 treatment groups was significantly higher than that of control group and 1.00 μmol•L^-1 group (P<0.05), but had no significant difference with that of 0.25 μmol•L^-1 group (P>0.05). In conclusion,high concentration of 5-Aza-CdR have side effect on the proliferation and chromosome ploidy of Debao pig fibroblasts. Applying low concentration (≤0.25 μmol•L^-1) of 5-Aza-CdR on donor cells reduced the methylation level of cloned embryos and consequently enhanced the developmental potency of handmade cloned pig embryos.

CLC Number:

LU Xing-rong，SUN Jun-ming，LI Zhi-peng，LI Lan-yu，XIE Bing-kun，LIU Qing-you，SHI De-shun，CUI Kui-qing. Effect of Methyltransferase Inhibitor 5-Aza-CdR on Development Potency of Handmade Cloning Embryo of Debao Pig[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2014, 45(8): 1274-1281.

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Effect of Methyltransferase Inhibitor 5-Aza-CdR on Development Potency of Handmade Cloning Embryo of Debao Pig

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