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种植体周围猪胚胎群体的特征和母体环境影响转录组谱。

Characteristics of peri-implantation porcine concepti population and maternal milieu influence the transcriptome profile.

出版信息

Mol Reprod Dev. 2010 Nov;77(11):978-89. doi: 10.1002/mrd.21253.

Abstract

Asynchrony of trophectoderm elongation, gestational days 11-12, is evident in porcine concepti, and rapid progression through this phase has been associated with conceptus competency. The goal of the current study was to determine the extent of transcriptomic responses of concepti to developmental delay and their physiological implications. Gestational day 11 concepti with the same morphology, ovoid and 7-8 mm, were isolated and designated as control or developmentally delayed if collected from a homogenous ovoid conceptus population or heterogeneous conceptus population (ovoid to filamentous), respectively. Total RNA prepared from four distinct control and four distinct developmentally delayed concepti, was analyzed using an Agilent high-density custom porcine microarray. Two hundred nine transcripts were found differentially expressed between normal and developmentally delayed concepti. Functional analysis of these genes indicated that a significant number of the genes regulate signal transduction/transcription, organismal development, metabolism, and cell adhesion and can be modulated by transforming growth factor β1 (TGFβ1). Ten genes were selected for real-time PCR validation of differential expression based on a known role in steroid synthesis, endometrium receptivity, and modulation of trophoblast differentiation/growth or interaction with TGFβ1. As in the microarray, all except one, achaete-scute complex homolog 2, were preferentially up-regulated in delayed concepti. Overall, findings suggested that despite similar morphology, the transcriptome of developmentally delayed concepti is distinct from control counterparts. Also highlighted were ways by which the conceptus' microenvironment might be affected and developmental factors that may be of interest to interrogate further to determine if, and how, they affect embryo competency/elongation.

摘要

滋养层延伸的异步性发生在 11-12 天的猪胚中,并且这一阶段的快速进展与胚泡的能力有关。本研究的目的是确定胚泡对发育延迟的转录组反应的程度及其生理意义。具有相同形态(椭圆形和 7-8mm)的 11 天猪胚,如果分别从均匀的椭圆形胚泡群体或不均匀的胚泡群体(椭圆形到丝状)中收集,则被指定为对照或发育延迟。从四个不同的对照和四个不同的发育延迟的胚泡中提取总 RNA,并使用 Agilent 高密度定制猪微阵列进行分析。在正常和发育延迟的胚泡之间发现了 209 个转录本存在差异表达。对这些基因的功能分析表明,相当数量的基因调节信号转导/转录、机体发育、代谢以及细胞黏附和 TGFβ1 可以调节这些基因。根据在类固醇合成、子宫内膜容受性以及调节滋养层分化/生长或与 TGFβ1 相互作用方面的已知作用,选择了 10 个基因进行实时 PCR 验证差异表达。与微阵列一样,除了 achaete-scute complex homolog 2 外,所有基因都在发育延迟的胚泡中优先上调。总体而言,研究结果表明,尽管形态相似,但发育延迟的胚泡的转录组与对照物不同。还强调了胚泡微环境可能受到影响的方式,以及可能进一步研究以确定它们是否以及如何影响胚胎能力/延伸的发育因素。

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