Cdc42 is implicated in polarity during meiotic resumption and blastocyst formation in the mouse

Cell division cycle 42 (Cdc42), a member of the Rho family of small guanosine triphosphatase (GTPase) proteins, regulates multiple cell functions, including: motility, proliferation, apoptosis, and cell morphology. In order to obtain insight into the role of Cdc42 in meiotic resumption and embryo development, we first evaluated its gene expression levels in mouse oocytes and embryos during in vitro development. Quantitative reverse transcription-polymerase chain reaction (RT-PCR) revealed high-expression levels in GV stage oocytes that steadily decreased up to the 2-cell (2C) stage embryo, and then expression increased during morulae and blastocyst formation. Indirect Immunocytochemistry also showed protein synthesis of CDC42 in the mouse oocytes and early embryos. Introducing small interference RNA (siRNA) of Cdc42 into germinal vesicle stage oocytes or zygotes specifically reduced both mRNA expression and protein synthesis of CDC42 in in vitro developed metaphase II oocytes and early embryos. Meiotic maturation and cytoskeleton assembly were significantly altered following siRNA injection into germinal vesicle stage oocytes. Injection of siRNA into the zygote did not affect cleavage or cell numbers in morulae, but significantly decreased in vitro development to the morula or blastocyst. These findings suggest that gene expression of Cdc42 is involved in meiotic resumption and blastocyst formation in the mouse, possibly through maintaining polarity.