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成纤维细胞生长因子受体在早期牛胚胎发育过程中的表达及其对滋养层细胞体外生长的作用的药理学分析。

The expression of fibroblast growth factor receptors during early bovine conceptus development and pharmacological analysis of their actions on trophoblast growth in vitro.

机构信息

Department of Animal Sciences, University of Florida, Gainesville, Florida 32611, USA.

出版信息

Reproduction. 2013 Jan 24;145(2):191-201. doi: 10.1530/REP-12-0220. Print 2013 Feb.

Abstract

The overall aim of this work was to examine the expression profiles for fibroblast growth factor receptors (FGFRs) and describe their biological importance during bovine pre- and peri-implantation conceptus development. FGFR1 and FGFR2 mRNAs were detected at 1-, 2-, 8-cell, morula and blastocyst stages whereas FGFR3 and FGFR4 mRNAs were detected after the 8-cell stage but not earlier. The abundance of FGFR1, FGFR3, and FGFR4 mRNAs increased at the morula and blastocyst stages. Immunofluorescence microscopy detected FGFR2 and FGFR4 exclusively in trophoblast cells whereas FGFR1 and FGFR3 were detected in both trophoblast cells and inner cell mass in blastocysts. Neither transcripts for FGF10 nor its receptor (FGFR2b) were temporally related to interferon τ (IFNT) transcript profile during peri- and postimplantation bovine conceptus development. A series of studies used a chemical inhibitor of FGFR kinase function (PD173074) to examine FGFR activation requirements during bovine embryo development. Exposing embryos to the inhibitor (1 μM) beginning on day 5 post-fertilization did not alter the percentage of embryos that developed into blastocysts or blastocyst cell numbers. The inhibitor did not alter the abundance of CDX2 mRNA but decreased (P<0.05) the relative abundance of IFNT mRNA in blastocysts. Exposing blastocysts to the inhibitor from days 8 to 11 post-fertilization reduced (P<0.05) the percentage of blastocysts that formed outgrowths after transfer to Matrigel-coated plates. In conclusion, each FGFR was detected in bovine embryos, and FGFR activation is needed to maximize IFNT expression and permit outgrowth formation.

摘要

本研究旨在探讨成纤维细胞生长因子受体(FGFRs)的表达谱,并描述其在牛胚前和植入前胚胎发育过程中的生物学重要性。FGFR1 和 FGFR2 mRNA 在 1-、2-、8-细胞、桑椹胚和囊胚阶段均有检测到,而 FGFR3 和 FGFR4 mRNA 则在 8-细胞阶段后被检测到,但在此之前未被检测到。FGFR1、FGFR3 和 FGFR4 mRNA 的丰度在桑椹胚和囊胚阶段增加。免疫荧光显微镜检测到 FGFR2 和 FGFR4 仅存在于滋养层细胞中,而 FGFR1 和 FGFR3 则存在于囊胚的滋养层细胞和内细胞团中。在牛胚前和植入后胚胎发育过程中,FGF10 及其受体(FGFR2b)的转录本与干扰素 τ(IFNT)的转录本均无时间相关性。一系列研究使用 FGFR 激酶功能的化学抑制剂(PD173074)来研究 FGFR 激活在牛胚胎发育过程中的要求。在受精后第 5 天开始向胚胎中添加抑制剂(1 μM),并不会改变胚胎发育成囊胚的百分比或囊胚细胞数量。抑制剂不会改变 CDX2 mRNA 的丰度,但会降低(P<0.05)囊胚中 IFNT mRNA 的相对丰度。在受精后第 8 至 11 天向囊胚中添加抑制剂,会降低(P<0.05)在 Matrigel 包被板上转移后形成外生体的囊胚的百分比。总之,在牛胚胎中检测到每种 FGFR,FGFR 的激活对于最大限度地提高 IFNT 的表达和允许外生体的形成是必需的。

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