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首页> 外文期刊>Reproductive Biology and Endocrinology >Temporal regulation of mRNAs for select bone morphogenetic proteins (BMP), BMP receptors and their associated SMAD proteins during bovine early embryonic development: effects of exogenous BMP2 on embryo developmental progression
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Temporal regulation of mRNAs for select bone morphogenetic proteins (BMP), BMP receptors and their associated SMAD proteins during bovine early embryonic development: effects of exogenous BMP2 on embryo developmental progression

机译：牛早期胚胎发育过程中某些骨形态发生蛋白（BMP），BMP受体及其相关SMAD蛋白的mRNA的时间调控：外源BMP2对胚胎发育进程的影响

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Background We previously demonstrated embryotrophic actions of maternal (oocyte-derived) follistatin during bovine early embryogenesis. Classical actions of follistatin are attributed to inhibition of activity of growth factors including activins and bone morphogenetic proteins (BMP). However, temporal changes in BMP mRNA in early bovine embryos and the effects of exogenous BMP on embryo developmental progression are not understood. The objectives of present studies were to characterize mRNA abundance for select BMP, BMP receptors and BMP receptor associated SMADs during bovine oocyte maturation and early embryogenesis and determine effects of addition of exogenous BMP protein on early development. Methods Relative abundance of mRNA for BMP2, BMP3, BMP7, BMP10, SMAD1, SMAD5, ALK3, ALK6, ALK2, BMPR2, ACVR2A and ACVR2B was determined by RT-qPCR analysis of germinal vesicle (GV) and in vitro matured metaphase II (MII) oocytes and in vitro produced embryos collected at pronuclear, 2-cell (C), 4C, 8C, 16C, morula and blastocyst stages. Effects of addition of recombinant human BMP2 (0, 1, 10 and 100?ng/ml) during initial 72?h of embryo culture on early cleavage (within 30?h post insemination), total cleavage, development to 8C-16C and blastocyst stages and blastocyst mRNA abundance for markers of inner cell mass (NANOG) and trophectoderm (CDX2) were also determined. Results Abundance of mRNA for BMP2, BMP10, SMAD1, SMAD5, ALK3, ALK2, BMPR2 and ACVR2B was elevated in MII oocytes and/or pronuclear stage embryos (relative to GV) and remained elevated through the 8C -16C stages, whereas BMP3, BMP7 and ALK2 mRNAs were transiently elevated. Culture of embryos to the 8C stage in the presence of α-amanitin resulted in increased abundance for all of above transcripts examined relative to untreated 8C embryos. Effects of addition of exogenous BMP2 on early cleavage rates and rates of development to 8C-16C and blastocyst stages were not observed, but BMP2 treatment increased blastocyst mRNA for CDX2 and NANOG. Conclusions Abundance of maternally derived mRNAs for above BMP system components are dynamically regulated during oocyte maturation and early embryogenesis. Exogenous BMP2 treatment does not influence progression to various developmental endpoints, but impacts characteristics of resulting blastocysts. Results support a potential role for BMPs in bovine early embryogenesis.
机译：背景我们以前证明了母体（卵母细胞）卵泡抑素在牛早期胚胎发生过程中的胚胎营养作用。卵泡抑素的经典作用归因于抑制包括激活素和骨形态发生蛋白（BMP）在内的生长因子的活性。然而，尚不清楚早期牛胚胎中BMP mRNA的时间变化以及外源BMP对胚胎发育进程的影响。本研究的目的是鉴定牛卵母细胞成熟和早期胚胎发生过程中所选BMP，BMP受体和与BMP受体相关的SMAD的mRNA丰度，并确定添加外源BMP蛋白对早期发育的影响。方法通过生胚囊泡（GV）和体外成熟中期II（MII）的RT-qPCR分析确定BMP2，BMP3，BMP7，BMP10，SMAD1，SMAD5，ALK3，ALK6，ALK2，BMPR2，ACVR2A和ACVR2B的mRNA相对丰度）卵母细胞和体外产生的胚胎在原核，2细胞（C），4C，8C，16C，桑ula和胚泡阶段收集。在胚胎培养的最初72小时内添加重组人BMP2（0、1、10和100μng/ ml）对早期卵裂（授精后30μh内），总卵裂，发育至8C-16C和胚泡的影响还确定了内细胞团（NANOG）和滋养外胚层（CDX2）标记物的阶段和胚泡mRNA丰度。结果MII卵母细胞和/或核前期胚胎（相对于GV）中BMP2，BMP10，SMAD1，SMAD5，ALK3，ALK2，BMPR2和ACVR2B的mRNA丰度升高，并且在8C -16C阶段仍升高，而BMP3，BMP7和ALK2 mRNA瞬时升高。与未经处理的8C胚胎相比，在存在α-amanitin的情况下将胚胎培养到8C阶段会导致上述所有转录物的丰度增加。没有观察到添加外源BMP2对早期卵裂率和向8C-16C和胚泡阶段发育的影响，但是BMP2处理增加了CDX2和NANOG胚泡mRNA的表达。结论在卵母细胞成熟和早期胚胎发生过程中，上述BMP系统组成部分的母体来源mRNA的存在是动态调节的。外源BMP2处理不影响进展至各种发育终点，但会影响所产生的胚泡的特征。结果支持BMPs在牛早期胚胎发生中的潜在作用。

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