Block Jeremy, Wrenzycki Christine, Niemann Heiner, Herrmann Doris, Hansen Peter J
Department of Animal Sciences, University of Florida, Gainesville, Florida 32611, USA.
Mol Reprod Dev. 2008 May;75(5):895-903. doi: 10.1002/mrd.20826.
Addition of insulin-like growth factor-1 (IGF-1) to culture medium increases the proportion of bovine embryos that develop to the blastocyst stage and increases embryo survival following transfer to heat-stressed, lactating dairy cows. The objective of the present study was to determine molecular and cellular correlates of these actions of IGF-1. Embryos were produced in vitro and cultured for 7 days with or without 100 ng/ml IGF-1. On d 7 after insemination, grade 1 expanded blastocysts were harvested and used to determine total cell number, percent apoptosis, cell allocation to the inner cell mass and trophectoderm, and the relative abundance of several developmentally important gene transcripts. There was no significant effect of IGF-1 treatment on blastocyst cell number, the proportion of blastomeres that were apoptotic, or the number of cells in the inner cell mass and trophectoderm. However, differences in the relative abundance of several mRNA transcripts were observed between control and IGF-1 treated embryos. Addition of IGF-1 increased (P < 0.02) amounts of mRNA for IGF binding protein-3 and desmocollin II and tended (P < 0.08) to increase amounts of mRNA for Na/K ATPase and Bax. Moreover, IGF-1 treatment decreased (P < 0.05) steady-state amounts of transcripts for heat shock protein 70 and tended (P < 0.08) to reduce amounts of IGF-1 receptor mRNA. In conclusion, increased survival of embryos treated with IGF-1 does not appear due to effects on cell number, percent apoptosis, or cell allocation. Addition of IGF-1 to culture can, however, alter expression of several transcripts which may be important for embryo development and survival following transfer.
在培养基中添加胰岛素样生长因子-1(IGF-1)可提高发育至囊胚阶段的牛胚胎比例,并提高将胚胎移植到热应激泌乳奶牛后其存活的几率。本研究的目的是确定IGF-1这些作用的分子和细胞相关性。胚胎在体外产生,并在添加或不添加100 ng/ml IGF-1的情况下培养7天。在授精后第7天,收集1级扩张囊胚,用于确定总细胞数、凋亡百分比、内细胞团和滋养外胚层的细胞分配,以及几种对发育重要的基因转录本的相对丰度。IGF-1处理对囊胚细胞数、凋亡的卵裂球比例或内细胞团和滋养外胚层中的细胞数没有显著影响。然而,在对照胚胎和经IGF-1处理的胚胎之间观察到几种mRNA转录本相对丰度的差异。添加IGF-1可使IGF结合蛋白-3和桥粒芯糖蛋白II的mRNA量增加(P<0.02),并使钠钾ATP酶和Bax的mRNA量有增加趋势(P<0.08)。此外,IGF-1处理可使热休克蛋白70转录本的稳态量减少(P<0.05),并使IGF-1受体mRNA量有减少趋势(P<0.08)。总之,用IGF-1处理的胚胎存活率提高似乎不是由于对细胞数、凋亡百分比或细胞分配的影响。然而,在培养基中添加IGF-1可改变几种转录本的表达,这可能对胚胎发育和移植后的存活很重要。