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首页> 外文期刊>Molecular reproduction and development >Involvement of ER-calreticulin-Ca2+ signaling in the regulation of porcine oocyte meiotic maturation and maternal gene expression.
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Involvement of ER-calreticulin-Ca2+ signaling in the regulation of porcine oocyte meiotic maturation and maternal gene expression.

机译：ER-钙网蛋白-Ca 2 + 信号参与猪卵母细胞减数分裂成熟和母体基因表达的调控。

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Calcium is one of the most ubiquitous signaling molecules, and controls a wide variety of cellular processes. It is mainly stored in the endoplasmic reticulum (ER), bound to lumenal proteins. Calreticulin is the major Ca2+-binding chaperone in oocytes, and is integral to numerous cellular functions. To better understand the role of the ER-calreticulin-Ca2+ pathway in oocyte maturation and early embryogenesis, we characterized the porcine calreticulin gene and investigated its expression profile during oocyte maturation and early embryonic development. Calreticulin was widely expressed in pig tissues and its transcripts were downregulated during maturation, especially at 44 hr, and were undetectable at the blastocyst stage. We also investigated the effect of increased cytosolic Ca2+ induced by the Ca2+-ATPase inhibitor, cyclopiazonic acid (CPA), on pig oocyte maturation and maternal gene expression. CPA at 10 micro M did not inhibit germinal vesicle breakdown, but did result in the arrest of 38.6% oocytes at or before the MI stage. In addition, expression of the maternal genes C-mos, BMP15, GDF9, and Cyclin B1 was significantly increased in CPA-treated MII oocytes compared with control groups. These data were supported by the results of poly(A)-test PCR, which revealed that the cyclin B1 short isoform (CB-S), GDF9, and C-mos underwent more intensive polyadenylation modification in CPA-treated oocytes than control oocytes, suggesting that polyadenylation may influence Ca2+-modulated changes in gene expression. Furthermore, CPA treatment decreased the percentage of four-cell parthenotes that developed into blastocysts, suggesting the need for functional SR/ER Ca2+-ATPase pumps or Ca2+ signals during early embryo development after zygotic genome activation. Together, these data indicate that ER-calreticulin-associated Ca2+ homeostasis plays a role in oocyte and embryo development, and that alterations in maternal gene expression may contribute to the underlying molecular mechanism, at least partially, via polyadenylation.
机译：钙是最普遍存在的信号分子之一，它控制着各种各样的细胞过程。它主要存储在与内腔蛋白结合的内质网（ER）中。钙网蛋白是卵母细胞中主要的Ca 2 + 结合伴侣蛋白，是众多细胞功能的组成部分。为了更好地了解ER-钙网蛋白-Ca 2 + 途径在卵母细胞成熟和早期胚胎发生中的作用，我们对猪钙网蛋白基因进行了表征，并研究了其在卵母细胞成熟和早期胚胎发育中的表达情况。钙网蛋白在猪组织中广泛表达，其转录物在成熟过程中尤其是在第44小时被下调，并且在胚泡期无法检测到。我们还研究了Ca 2 + -ATPase抑制剂环吡嗪酸（CPA）诱导的胞质Ca 2 + 升高对猪卵母细胞成熟和母体基因表达的影响。 CPA在10 micro M时不会抑制生小泡的破裂，但是确实导致了MI期或之前的38.6％卵母细胞停滞。此外，与对照组相比，经CPA处理的MII卵母细胞中母本基因C-mos，BMP15，GDF9和Cyclin B1的表达显着增加。这些数据得到了poly（A）-test PCR结果的支持，该结果表明，与对照卵母细胞相比，在CPA处理的卵母细胞中，细胞周期蛋白B1短异构体（CB-S），GDF9和C-mos经历了更强烈的聚腺苷酸化修饰，提示聚腺苷酸化可能影响Ca 2 + 调节的基因表达变化。此外，CPA处理降低了发育为胚泡的四细胞单性生殖的百分比，表明需要功能性SR / ER Ca 2 + -ATPase泵或Ca 2 + 信号合子基因组激活后早期胚胎发育过程中。总之，这些数据表明，ER-钙网蛋白相关的Ca 2 + 稳态在卵母细胞和胚胎发育中起作用，并且母体基因表达的改变可能至少部分地参与了潜在的分子机制，通过聚腺苷酸化。

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