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电场强度对体细胞核移植获得的家猫胚胎融合及体外发育的影响

Effects of electric field strengths on fusion and in vitro development of domestic cat embryos derived by somatic cell nuclear transfer.

作者信息

Karja Ni Wayan Kurniani, Otoi Takeshige, Wongsrikeao Pimprapar, Shimizu Ryohei, Murakami Masako, Agung Budiyanto, Fahrudin Mokhamad, Nagai Takashi

机构信息

Laboratory of Animal Reproduction, The United Graduate School of Veterinary Science, Yamaguchi University, 1677-1 Yoshida, Yamaguchi 753-8515, Japan.

出版信息

Theriogenology. 2006 Sep 15;66(5):1237-42. doi: 10.1016/j.theriogenology.2006.03.034. Epub 2006 May 3.

Abstract

The present study was conducted to determine the effect of electric field strength on the rate of membrane fusion between the somatic cell and cytoplast and on subsequent in vitro development of reconstructed embryos. Additionally, the in vitro developmental competence of cat oocytes artificially activated after 44 h of maturation culture was examined. An efficient fusion rate (64.2%) was obtained by applying a single pulse of 1.5 kV/cm for 50 micros, and the fusion rate remained almost constant at the higher field intensity (59.8 and 54.9% at 1.7 and 2.0 kV/cm, respectively). Although the cleavage rate of fused embryos increased with an increase of the electric field strength, there were no differences among the groups with respect to the proportion of development to the morula and blastocyst stages. In the additional experiment, oocytes at the metaphase II stage after culture for 44 h were activated by the combination of calcium ionophore (CaI) with cycloheximide (CHX). Some (11.8%) of activated oocytes developed to the blastocyst stage. Results from this study indicated that electric field strength affects the rates of fusion and cleavage but has no significant effects on the development to the blastocyst stage of reconstructed embryos. Prolonged maturation culture of cat oocytes (up to 44 h) decreased their ability to develop to the blastocyst stage.

摘要

本研究旨在确定电场强度对体细胞与细胞质体之间膜融合速率以及对重构胚胎随后体外发育的影响。此外,还检测了成熟培养44小时后人工激活的猫卵母细胞的体外发育能力。通过施加1.5 kV/cm的单脉冲50微秒,获得了较高的融合率(64.2%),在更高的场强下(1.7 kV/cm和2.0 kV/cm时分别为59.8%和54.9%),融合率几乎保持不变。虽然融合胚胎的分裂率随着电场强度的增加而提高,但在发育到桑椹胚和囊胚阶段的比例方面,各实验组之间没有差异。在额外的实验中,培养44小时处于中期II期的卵母细胞通过钙离子载体(CaI)与放线菌酮(CHX)联合激活。一些(11.8%)激活的卵母细胞发育到了囊胚阶段。本研究结果表明,电场强度影响融合率和分裂率,但对重构胚胎发育到囊胚阶段没有显著影响。猫卵母细胞延长成熟培养(长达44小时)会降低其发育到囊胚阶段的能力。

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