Akoury Elie, Zhang Li, Ao Asangla, Slim Rima
Department of Human Genetics, McGill University Health Center, Montreal, QC, Canada Department of Obstetrics and Gynecology, McGill University Health Center, Montreal, QC, Canada.
Department of Obstetrics and Gynecology, McGill University Health Center, Montreal, QC, Canada.
Hum Reprod. 2015 Jan;30(1):159-69. doi: 10.1093/humrep/deu291. Epub 2014 Oct 29.
What is the subcellular localization in human oocytes and preimplantation embryos, of the two maternal-effect proteins, NLRP7 and KHDC3L, responsible for recurrent hydatidiform moles (RHMs)?
NLRP7 and KHDC3L localize to the oocyte cytoskeleton and are polar and absent from the cell-to-cell contact region in early preimplantation embryos.
NLRP7 and KHDC3L expression has been described at the RNA level in some stages of human oocytes and preimplantation embryos and at the protein level by immunohistochemistry in human and bovine ovaries. NLRP7 and KHDC3L co-localize to the microtubule organizing center and/or the Golgi apparatus in human hematopoietic cells.
STUDY DESIGN, SIZE, DURATION: A total of 164 spare human oocytes and embryos from patients undergoing in vitro fertilization were used.
PARTICIPANTS/MATERIALS, SETTING, METHODS: Oocytes and early cleavage-stage embryos were fixed, immunostained with NLRP7 and/or KHDC3L antibodies, and analyzed using high-resolution confocal immunofluorescence and electron microscopies.
NLRP7 and KHDC3L localize to the cytoskeleton and are predominant at the cortical region in growing oocytes. After the first cellular division, these two maternal-effect proteins become asymmetrically confined to the outer cortical region and excluded from the cell-to-cell contact region until the blastocyst stage where NLRP7 and KHDC3L homogeneously redistribute to the cytoplasm and the nucleus, respectively.
LIMITATIONS, REASONS FOR CAUTION: We could not analyze fresh human oocytes and embryos. The analyzed materials were donated by patients undergoing assisted reproductive technologies and released for research 1-3 days after their collection and the transfer of embryos to the patients.
Our study is the first comprehensive and high-resolution localization of the only two known maternal-effect proteins, NLRP7 and KHDC3L, in human oocytes and preimplantation embryos. Our data contribute to a better understanding of the roles of these two proteins in the integrity of the oocytes, post-zygotic divisions, and cell-lineage differentiation.
STUDY FUNDING/COMPETING INTERESTS: This work was supported by the Canadian Institute of Health Research (86546 to R.S.); E.A. was supported by fellowships from the Research Institute of the McGill University Health Centre and a CREATE award from the Réseau Québécois en Reproduction. All authors declare no conflict of interest.
导致复发性葡萄胎（RHMs）的两种母源效应蛋白NLRP7和KHDC3L在人类卵母细胞和植入前胚胎中的亚细胞定位是什么？
NLRP7和KHDC3L定位于卵母细胞的细胞骨架，在植入前早期胚胎中呈极性分布，且不存在于细胞间接触区域。
NLRP7和KHDC3L的表达已在人类卵母细胞和植入前胚胎的某些阶段通过RNA水平进行了描述，并通过免疫组织化学在人类和牛的卵巢中进行了蛋白质水平的描述。NLRP7和KHDC3L在人类造血细胞中共定位于微管组织中心和/或高尔基体。
研究设计、规模、持续时间：总共使用了164个来自接受体外受精患者的备用人类卵母细胞和胚胎。
参与者/材料、环境、方法：卵母细胞和早期卵裂期胚胎被固定，用NLRP7和/或KHDC3L抗体进行免疫染色，并使用高分辨率共聚焦免疫荧光和电子显微镜进行分析。
NLRP7和KHDC3L定位于细胞骨架，在生长中的卵母细胞的皮质区域占主导地位。在第一次细胞分裂后，这两种母源效应蛋白不对称地局限于外皮质区域，并被排除在细胞间接触区域之外，直到囊胚期，此时NLRP7和KHDC3L分别均匀地重新分布到细胞质和细胞核中。
局限性、谨慎的原因：我们无法分析新鲜的人类卵母细胞和胚胎。分析的材料由接受辅助生殖技术的患者捐赠，并在收集胚胎并将其移植给患者后1 - 3天释放用于研究。
我们的研究是首次对人类卵母细胞和植入前胚胎中仅有的两种已知母源效应蛋白NLRP7和KHDC3L进行全面且高分辨率的定位。我们的数据有助于更好地理解这两种蛋白质在卵母细胞完整性、合子后分裂和细胞谱系分化中的作用。
研究资金/利益冲突：这项工作得到了加拿大卫生研究院（给R.S.的86546号资助）；E.A.得到了麦吉尔大学健康中心研究所的奖学金以及魁北克生殖网络的CREATE奖。所有作者均声明无利益冲突。
