Peng Hui, Lin Xiujiao, Liu Fang, Wang Cheng, Zhang Wenchang
College of Animal Science, Fujian Agriculture and Forestry University, Fujian 350002, P. R. China.
J Reprod Dev. 2015;61(6):559-64. doi: 10.1262/jrd.2015-050. Epub 2015 Sep 28.
Nlrp9a, Nlrp9b and Nlrp9c are preferentially expressed in oocytes and early embryos in the mouse. Simultaneous genetic ablation of Nlrp9a and Nlrp9c does not affect early embryonic development, but the function of Nlrp9b in the process of oocyte maturation and embryonic development has not been elucidated. Here we show that both Nlrp9b mRNA and its protein are expressed in ovaries and the small intestine. Moreover, the NLRP9B protein was restricted to oocytes in the ovary and declined with oocyte aging. After ovulation and fertilization, NLRP9B protein was found in preimplantation embryos. Confocal microscopy demonstrated that it was mainly localized in the cytoplasm in the oocytes and blastomeres. Thus, this protein might play a role in oocyte maturation and early embryonic development. However, knockdown of Nlrp9b expression in GV-stage oocytes using RNA interference did not affect oocyte maturation or subsequent parthenogenetic development after Nlrp9b-deficient oocytes were activated. Furthermore, Nlrp9b knockdown zygotes could reach the blastocyst stage after being cultured for 3.5 days in vitro. These results provide the first evidence that the NLRP9B protein is dispensable for oocyte maturation and early embryonic development in the mouse.
Nlrp9a、Nlrp9b和Nlrp9c在小鼠的卵母细胞和早期胚胎中优先表达。Nlrp9a和Nlrp9c的同时基因敲除不影响早期胚胎发育,但Nlrp9b在卵母细胞成熟和胚胎发育过程中的功能尚未阐明。在此我们表明,Nlrp9b mRNA及其蛋白在卵巢和小肠中均有表达。此外,NLRP9B蛋白局限于卵巢中的卵母细胞,并随着卵母细胞老化而减少。排卵和受精后,在植入前胚胎中发现了NLRP9B蛋白。共聚焦显微镜显示,它主要定位于卵母细胞和卵裂球的细胞质中。因此,该蛋白可能在卵母细胞成熟和早期胚胎发育中发挥作用。然而,使用RNA干扰敲低GV期卵母细胞中Nlrp9b的表达,在Nlrp9b缺陷型卵母细胞激活后,并不影响卵母细胞成熟或随后的孤雌生殖发育。此外,Nlrp9b敲低的受精卵在体外培养3.5天后可发育至囊胚期。这些结果首次证明,NLRP9B蛋白在小鼠卵母细胞成熟和早期胚胎发育中并非必需。