ORIGINAL ARTICLE Reproductive biology NLRP7 and KHDC3L, the two maternal- effect proteins responsible for recurrent hydatidiform moles, co-localize to the oocyte cytoskeleton Elie Akoury 1,2 , Li Zhang 2 , Asangla Ao 2 , and Rima Slim 1,2, * 1 Department of Human Genetics, McGill University Health Center, Montreal, QC, Canada 2 Department of Obstetrics and Gynecology, McGill University Health Center, Montreal, QC, Canada *Correspondence address. Montreal General Hospital Research Institute, L3-121, 1650 Cedar Ave., Montreal, Quebec, Canada H3G 1A4. Tel: +1-514-934-1934 (ext.44550); Fax: +1-514-934-8261; E-mail: rima.slim@muhc.mcgill.ca Submitted on July 7, 2014; resubmitted on September 11, 2014; accepted on October 9, 2014 study question: 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)? summaryanswer: NLRP7 and KHDC3L localize to the oocyte cytoskeleton and are polar and absent from the cell-to-cell contact region in early preimplantation embryos. what is known already: 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. main results and the role of chance: 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. wider implications of the findings: 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 interest(s): 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 Re ´seau Que ´be ´cois en Reproduction. All authors declare no conflict of interest. Key words: NLRP7 / KHDC3L / recurrent hydatidiform mole / oocyte cortex / oocyte cytoskeleton & The Author 2014. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oup.com Human Reproduction, Vol.0, No.0 pp. 1 –11, 2014 doi:10.1093/humrep/deu291 Hum. Reprod. Advance Access published October 29, 2014 by guest on October 30, 2014 http://humrep.oxfordjournals.org/ Downloaded from