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ghrelin 对体外受精绵羊胚胎发育能力和基因表达的影响。

Effects of ghrelin on developmental competence and gene expression of in vitro fertilized ovine embryos.

机构信息

College of Animal Sciences of Zhejiang University, Key Laboratory for Molecular Animal Nutrition, Ministry of Education, Hangzhou, PR China.

出版信息

Theriogenology. 2013 Mar 1;79(4):695-701. doi: 10.1016/j.theriogenology.2012.11.026. Epub 2013 Jan 2.

Abstract

The objective was to determine the effects of various ghrelin concentrations (0, 10, 50, and 250 ng/mL) during in vitro oocyte maturation (IVM) and in vitro embryo culture (IVC) on ovine embryo development and expression patterns of genes involved in transcription regulation (OCT4), glucose transport (GLUT1), pregnancy recognition (IFNT), and ghrelin receptor (GHSR-1a). Rates of cleavage and blastocyst formation were decreased when oocytes were matured with 250 ng/mL ghrelin compared with 0, 10, or 50 ng/mL ghrelin (P < 0.05). Addition of 50 ng/mL ghrelin during IVC or during IVM and IVC significantly increased blastocyst rates (35.3% and 36.7% vs. 26.9% and 26.8%, respectively) and total cell numbers per blastocyst (110.4 and 108.3 vs. 81.2 and 77.4) compared with 0 and 10 ng/mL ghrelin. However, a high concentration (250 ng/mL) of ghrelin during IVM and IVC decreased cleavage, blastocyst rate, and total cell number of blastocyst compared with low concentrations (P < 0.05). Relative abundances of GLUT1 and IFNT transcripts were higher in blastocysts treated with 50 ng/mL ghrelin during IVC compared with other concentrations (P < 0.05). Expression of GHSR-1a was higher when 10 ng/mL ghrelin was added during IVM (0.079) or during IVM and IVC (0.053) compared with other treatments. However, addition of ghrelin at higher concentrations (50 or 250 ng/mL) reduced relative abundances of GHSR-1a transcripts (0.032 and 0.039, P < 0.05). In conclusion, appropriate concentrations of ghrelin promoted ovine blastocyst formation in vitro and increased expression of GLUT1, IFNT, and GHSR-1a genes in blastocysts, although a high concentration of ghrelin suppressed embryo development.

摘要

目的是确定不同浓度的生长激素释放肽(0、10、50 和 250ng/ml)在体外卵母细胞成熟(IVM)和体外胚胎培养(IVC)过程中对绵羊胚胎发育和参与转录调节的基因表达模式(OCT4)、葡萄糖转运(GLUT1)、妊娠识别(IFNT)和生长激素释放肽受体(GHSR-1a)的影响。与 0、10 或 50ng/ml 生长激素释放肽相比,用 250ng/ml 生长激素释放肽成熟的卵母细胞的卵裂和囊胚形成率降低(P < 0.05)。在 IVC 或 IVM 和 IVC 期间添加 50ng/ml 生长激素释放肽显著增加囊胚率(35.3%和 36.7%与 26.9%和 26.8%,分别)和每个囊胚的总细胞数(110.4 和 108.3 与 81.2 和 77.4)与 0 和 10ng/ml 生长激素释放肽。然而,在 IVM 和 IVC 期间高浓度(250ng/ml)的生长激素释放肽与低浓度(P < 0.05)相比,降低了卵裂、囊胚率和囊胚的总细胞数。与其他浓度相比,在 IVC 期间用 50ng/ml 生长激素释放肽处理的囊胚中 GLUT1 和 IFNT 转录本的相对丰度更高(P < 0.05)。当在 IVM 期间添加 10ng/ml 生长激素释放肽(0.079)或在 IVM 和 IVC 期间添加 10ng/ml 生长激素释放肽(0.053)时,GHSR-1a 的表达高于其他处理。然而,添加更高浓度的生长激素释放肽(50 或 250ng/ml)降低了 GHSR-1a 转录本的相对丰度(0.032 和 0.039,P < 0.05)。总之,适当浓度的生长激素释放肽促进绵羊囊胚体外形成,并增加囊胚中 GLUT1、IFNT 和 GHSR-1a 基因的表达,尽管高浓度的生长激素释放肽抑制胚胎发育。

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