Functional analysis of microRNA pathway genes in the somatic gonad and germ cells during ovulation in C. elegans.
Functional analysis of microRNA pathway genes in the somatic gonad and germ cells during ovulation in C. elegans.
复制标题
线虫排卵期间体细胞性腺和生殖细胞中 microRNA 途径基因的功能分析。
DOI：
10.1016/j.ydbio.2017.04.007
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发表时间：
2017
影响因子：
2.7
通讯作者：
Abbott,AllisonL
中科院分区：
文献类型：
--
作者：
Rios,Carmela;Warren,David;Olson,Benjamin;Abbott,AllisonL
MicroRNAs(MiRNAs)是基因表达的转录后调节因子，在动物发育和生理中发挥重要作用，但大多数miRNAs的功能尚不清楚。由于DROSHA或Pasha/Dgcr8活性丧失而导致miRNA生物合成减少的虫子是不育的，无法排卵，这表明miRNAs是卵母细胞成熟和排卵过程中所必需的。从这种渗透性无菌表型开始，使用创建的新菌株进行组织特异性RNAi，我们表征了C。Pasha、pash-1和两个miRNA特异的Argane，alg-1和alg-2，在体细胞和生殖细胞中参与排卵的调节。PASH-1活性的条件性丧失导致排卵率降低以及基础和排卵鞘收缩。同样，组织特异性RNAi在体细胞性腺细胞中敲除miRNA特异的Argavies会导致排卵率降低，并导致基础和排卵鞘收缩。MiRNA途径基因活性降低导致一系列缺陷，包括在alg-1RNAi后观察到的排卵事件中，约42%的卵母细胞在进入受精膜远端时被夹住。在暴露于对照RNAi的蠕虫上没有观察到这种表型。相反，在生殖细胞中敲除alg-1和alg-2会导致很少的卵母细胞成熟和排卵缺陷。这些数据确定了排卵过程中需要体细胞性腺细胞中miRNA途径基因活性的特定步骤。
MicroRNAs (miRNAs) are post-transcriptional regulators of gene expression that play critical roles in animal development and physiology, though functions for most miRNAs remain unknown. Worms with reduced miRNA biogenesis due to loss of Drosha or Pasha/DGCR8 activity are sterile and fail to ovulate, indicating that miRNAs are required for the process of oocyte maturation and ovulation. Starting with this penetrant sterile phenotype and using new strains created to perform tissue specific RNAi, we characterized the roles of theC. elegansPasha,pash-1,and two miRNA-specific Argonautes,alg-1andalg-2,in somatic gonad cells and in germ cells in the regulation of ovulation. Conditional loss ofpash-1activity resulted in a reduced rate of ovulation and in basal and ovulatory sheath contractions. Similarly, knockdown of miRNA-specific Argonautes in the cells of the somatic gonad by tissue-specific RNAi results in a reduction of the ovulation rate and in basal and ovulatory sheath contractions. Reduced miRNA pathway gene activity resulted in a range of defects, including oocytes that were pinched upon entry of the oocyte into the distal end of the spermatheca in about 42% of the ovulation events observed followingalg-1RNAi. This phenotype was not observed on worms exposed to control RNAi. In contrast, knockdown ofalg-1andalg-2in germ cells results in few defects in oocyte maturation and ovulation. These data identify specific steps in the process of ovulation that require miRNA pathway gene activity in the somatic gonad cells.