CHARACTERIZATION OF ZYGOTE ARREST ONE (ZAR1) IN NONHUMAN PRIMATES

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. ZAR1 is a newly identified oocyte-expressed protein conserved during evolution. Female mice lacking ZAR1 are infertile due to a block of embryogenesis at the zygote stage. Thus, ZAR1 belongs to a small group of maternal-effect proteins that are required for the oocyte-to-embryo transition in mice. In humans, however, ZAR1 mRNA is detectable in both the ovary and the testis. To elucidate functions of ZAR1 in human fertility, we chose nonhuman primates as our mammalian system due to their close resemblance to humans in development. The aim of this project is to demonstrate potential functions of ZAR1 during early embryogenesis and identify key molecules that function at the oocyte-to-embryo transition in monkeys. We have discovered a monkey cDNA that is highly homologous to mouse and human ZAR1 gene, and determined the expression pattern of monkey ZAR1 mRNA. The specific aims are as follows: (1) to characterize ZAR1 protein expression in monkeys. Specific antibodies that recognize a highly conserved region of ZAR1 protein will be generated and used to determine ZAR1 expression in monkeys and humans; (2) to demonstrate potential functions of ZAR1 during early embryo development in monkeys. In vitro RNAi and transgenic techniques will be employed in monkey oocytes and early embryos; (3) to identify key molecules that function at the oocyte-to-embryo transition in nonhuman primates. We will perform microarray analysis to identify differentially expressed molecules in ZAR1 knockdown embryos. The study described here is among the first attempts to illustrate molecular mechanisms that regulate preimplantation embryogenesis in nonhuman primates.