Expression of retinol‐binding protein messenger RNA and retinoic acid receptors in preattachment bovine embryos
Expression of retinol‐binding protein messenger RNA and retinoic acid receptors in preattachment bovine embryos
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视黄醇结合蛋白信使RNA和视黄酸受体在附着前牛胚胎中的表达
DOI：
10.1002/mrd.1090
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发表时间：
2001
影响因子：
2.5
通讯作者：
G. Morgan
中科院分区：
文献类型：
--
作者：
M. Mohan;J. Malayer;R. Geisert;G. Morgan
在牛中，视黄酸 (RA) 与胚胎的发育潜力间接相关。 RA 由视黄醇结合蛋白 (RBP) 转运，RA 的作用由核视黄酸受体 (RAR) 的几种亚型介导。牛胚胎是从当地屠宰场收集的卵巢中采集的卵母细胞在体外产生的，在卵母细胞、2、4、8、16 至 20 细胞、桑椹胚、囊胚和孵化囊胚阶段冷冻在液氮中。采用逆转录聚合酶链反应 (RT-PCR)，我们研究了 RBP、RARα、RARβ、RARγ 和 3-磷酸甘油醛脱氢酶 (GAPDH) 的 mRNA 表达。从每个阶段的 25 个合并胚胎中提取总 RNA，并重复 RT-PCR 分析三次。在所有阶段均检测到 GAPDH 转录本。从卵母细胞到孵化囊胚的所有阶段也检测到了 RBP、RARα 和 RARγ 的转录本。在任何阶段均未检测到RARβ的表达。使用针对 RARα 和 RARγ2 的多克隆抗体对完整和孵化的囊胚进行整体免疫组织化学，以研究这些胚胎 mRNA 是否被翻译为成熟蛋白。在完整和孵化的囊胚的内细胞团和滋养外胚层中观察到 RARα 和 RARγ2 的强烈免疫染色。牛胚胎中 RBP、RARα、RARγ 以及 RARα 和 RARγ2 受体蛋白 mRNA 的表达表明，RA 可能在该物种的植入前发育过程中直接调节基因表达。摩尔。重现。开发。 60: 289–296, 2001。© 2001 Wiley-Liss, Inc.
In cattle, retinoic acid (RA) has been indirectly associated with developmental potential of the embryo. RA is transported by retinol‐binding protein (RBP) and actions of RA are mediated by several subtypes of nuclear retinoic acid receptors (RAR). Bovine embryos, produced in vitro from oocytes harvested from ovaries collected at a local abattoir, were frozen in liquid nitrogen at the oocyte, 2‐, 4‐, 8‐, 16 to 20‐cell, morula, blastocyst, and hatched blastocyst stages. Employing reverse transcription polymerase chain reaction (RT‐PCR) we investigated mRNA expression for RBP, RARα, RARβ, RARγ, and glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH). Total RNA was extracted from 25 pooled embryos at each stage and RT‐PCR analysis was repeated thrice. GAPDH transcript was detected in all stages. Transcripts for RBP, RARα, and RARγ were also detected in all stages from the oocyte through to the hatched blastocyst. Expression of RARβ was not detected at any stage. Whole‐mount immunohistochemistry was performed with intact and hatched blastocysts using polyclonal antibodies against RARα and RARγ2 to investigate if these embryonic mRNAs were translated to the mature protein. Strong immunostaining was observed for both RARα and RARγ2 in the inner cell mass and trophectoderm of intact and hatched blastocysts. Expression of mRNA for RBP, RARα, RARγ, and of the RARα and RARγ2 receptor proteins in the bovine embryo suggests that RA is likely to directly regulate gene expression during preimplantation development in that species. Mol. Reprod. Dev. 60: 289–296, 2001. © 2001 Wiley‐Liss, Inc.
影响因子：
4.2
作者：
NAPOLI, JL
通讯作者：
NAPOLI, JL