Research On The Roles Of P97 During Mouse Oocytes Meiotic Maturation

p97 also called valosin-containing protein(VCP) in mammals,Cdc48 p in yeast, or CDC-48 in Caenorhabditis elegans—is well conserved among all eukaryotes and is essential for growth in Saccharomyces cerevisiae, Drosophila melanogaster, C. elegans and mice. p97 is highly abundant, accounting for about 1% of the total cellular protein.It has been suggested that p97 is a ubiquitin-selective chaperone and that its key function is to unfold proteins and disassemble protein complexes. p97 converts chemical energy generated from ATP hydrolysis into mechanical force used for conformational changes of target proteins as a unfoldase or a segregase.p97 plays critical roles in a broad range of diverse cellular processes, including Golgi, endoplasmic reticulum(ER), and nuclear membrane reassembly, ER associated degradation(ERAD) the ubiquitin-proteasome system, cell cycle regulation, DNA repair, preventing polyglutamine aggregation, autophagosome maturation, and mitophagy. Functional diversity of p97 is mainly determined by differential binding of distinct cofactors/adaptor proteins.In mitosis, p97 was reported to play roles in DNA replication, Aurora B kinase localization, chromosome segregation, microtubule spindle disassembly, and nuclear envelope reformation。p97 is a multifunctional protein and its functions are required for cell viability. However, its precise role and mechanism for meiosis maturation remain elusive.This study aims to explore the roles of p97 during mouse oocytes meiotic maturation.Main innovation point of this study: using immunofluorescence staining to confirm that p97 plays an important role in the meiotic maturation of mouse oocyte. Objective:To explore the roles of p97 during mouse oocytes meiotic maturation. Methods:1.Western Blot is used assay expression levels of p97 protein. Immunofluorescence staining is applied to detect the expression of p97 in all stages of meiosis in the oocyte and is used to analysis whether meiotic maturation of oocyte is different between of the group treated with DBe Q(inhibitor of p97 function). Immunofluorescence staining detects the changes of expression of CDC25 a and CDC25 b in oocytes after DBe Q treatment.2.Immunofluorescence staining is also used to detect whether there are abnormal chromosome separation in the oocytes which are treated with DBe Q. Chromosomal spread technology is used to detect the changes of number and structure of chromosomes. Results:1.The dysfunction of p97 in oocyte blocks GVBD and meiosis progression after GVBD, therefore oocyte meiotic process arrests at MI or ATI.2. The expression level of CDC25 a, CDC25 b of nuclear decreases in the oocytes treated after DBe Q.3.Abnormal chromatin condensation,chromosomal disorganized and chromosomes separated abnormally will be detected in the MI or ATI oocytes treated by DBe Q. Conclusion:1.p97 is essential for the recovery of meiosis progression in mouse oocytes.