The use of somatic cell nuclear transfer to obtain interspecific cloned embryos from brown brocket deer karyoplast and bovine cytoplast: Embryo development and nuclear gene expression

Luciana M. Melo , Sâmara B. Silva , Lívia C. Magalhães , Jenin V. Cortez , Satish Kumar , José M.B. Duarte , Luciana D. Rola , Maiana S. Chaves , Vicente J.F. Freitas
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引用次数: 1

Abstract

The brown brocket deer is a species of neotropical deer that is found in several regions of Brazil. Although not considered an endangered species, it can be used as a model for studies with somatic cell nuclear transfer (SCNT) in endangered species. Thus, this study aimed to verify the embryo development and nuclear gene expression in intraSCNT (bovine-bovine), interSCNT (deer-bovine) and in vitro fertilized (IVF) bovine embryos. Thus, from a culture of brown brocket deer fibroblasts and enucleated bovine oocytes, inter and intraspecific embryos were produced. In addition, a group of IVF bovine embryos was prepared. Quantitative PCR (qPCR) was performed from the blastocysts obtained from each experimental group to analyze the expression of OCT4, T-FAM, GJA1, BAX and BCL2. The blastocyst rate was higher (P < 0.05) in IVF group (44.0%) when compared to intraSCNT (11.3%) and inter SCNT (5.9%). The expression of BCL2 was close to null and for GJA1 was similar (P > 0.05) among experimental groups. In conclusion, bovine cytoplast was able to reprogram the brown brocket deer karyoplast and these cloned embryos were able to reach the blastocyst stage. The inverse relationship for BAX and BCL2 can indicate a high degree of apoptosis in blastocysts from interSCNT.

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利用体细胞核移植从褐小鹿核体和牛细胞质中获得种间克隆胚胎:胚胎发育和核基因表达
棕色小鹿是一种新热带鹿,在巴西的几个地区都有发现。虽然不被认为是濒危物种,但它可以作为研究濒危物种体细胞核移植(SCNT)的模型。因此,本研究旨在验证ascnt内(牛-牛)、scnt间(鹿-牛)和体外受精(IVF)牛胚胎的胚胎发育和核基因表达。因此,从褐小鹿成纤维细胞和去核牛卵母细胞的培养中,产生了种间和种内胚胎。此外,还制备了一组体外受精牛胚胎。采用定量PCR (qPCR)对各实验组囊胚进行OCT4、T-FAM、GJA1、BAX、BCL2的表达分析。囊胚率较高(P <0.05), IVF组(44.0%),而asnt组(11.3%)和SCNT组(5.9%)。BCL2的表达接近于零,GJA1的表达相似(P >0.05)。结果表明,牛细胞质能够对褐小鹿核体进行重编程,克隆的胚胎能够达到囊胚期。BAX和BCL2呈负相关,提示间充质干细胞囊胚高度凋亡。
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Theriogenology wild
Theriogenology wild Animal Science and Zoology, Veterinary Science
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