Localization Dynamics And Preliminary Research Function Of Ste-like Kinase (SLK) During Mouse Oocytes Maturation

Meiosis is the basis of Genetics, it guarantees the stability of chromosome number in the sexual reproduction of individual organisms and generations, it provides the genetic material to create variation in the process of sexual reproduction. Mouse oocyte meiosis is the extreme asymmetric division, very small polar body is excreted does not have the biological function and the egg contains the majority of maternal substance is excreted make preparations for the fertilization later and prophase embryonic development in the division process.The SLK (ste-20like kinase) is very conservative serine/threonine similar kinase in the evolution, is microtubule-associated protein, the downstream target proteins of Rho-GTPase Cdc42/Rac, involved in many important cellular activities, such as cell migration, cytoskeletal reorganization and apoptosis and affect cell cycle progression, it is necessary in mitosis.First of all, research the RNA expression levels of SLK Using RT-PCR amount of the GV phase, GVBD phase, MI phase and MⅡ phase in the mouse oocyte meiosis, that is cultured mouse oocytes at Oh,2h,8h and16h. The research showed the RNA expression levels of SLK at all times very high, and the RNA levels is gradually increasing with the meiosis process moves forward.Secondly, research the expression and localization of SLK in mouse oocytes during meiosis. Detected the expression and localization of SLK in mouse oocytes cultured in vitro Oh,2h,8h and16h, the corresponding mouse oocytes mature GV phase, GVBD phase, MI phase and MⅡ phase. Confirmed the SLK period expression in the mouse oocyte meiosis balanced the whole process. The result showed SLK located in the chromosome centromere By immunofluorescence staining in the meiosis.At the same time, after treat the oocyte by NOCO research the expression and localization of SLK in chromosome centromere by chromosome expansion. The experiments showed that the localization and expression of SLK after NOCO treatment was significantly stronger than the control (none-NOCO) in the MI phase and MⅡ phase of mouse oocyte meiosis. Experimental results show that the connection between microtubules protein and chromosome centromere was broke after the NOCO treatment, then the expression of SLK was significantly enhanced. The results showed that the SLK is microtubule-associated. SLK is very important for connection between the Spindle and chromosome.Then, research the localization relation between SLK and Crest in mouse oocytes during meiosis by Chromosome expansion. Experiments showed that the SLK expression at the entire process of meiosis in mouse oocytes,and in the Pro-MI, MI and MⅡ was mainly localized on chromosome centromeric.In addition the Co-transfection experiments show SLK was co-localization with Crest on chromosome centromere. The results show that SLK is associated with the checkpoint protein, we designed the next experiment to in order to further study What is the relationship SLK with the checkpoint protein. In order to verify the relationship of the SLK and checkpoint protein, research the localization between SLK and Mad1in mouse oocytes during meiosis using chromosome expansion. The experimental results show that SLK and Madl are co-local izetion on chromosome centromere in the Pro-MI phase of mouse oocytes. However, MAD1leave the centromere, while the SLK still present in the centromere in metaphase (MI). Mad1is back to the centromere coincides with the SLK after treat with NOCO. Experimental results show that the SLK has a close relationship with the checkpoint protein Mad1.Finally, in order to study the function of SLK in mouse oocytes, knock down the SLK injected with the Si-RNA and Morphlinor by microinjection in mouse oocytes, to observe the impact of research meiotic process lack of SLK.The experimental results show that the SLK hasn’t influence on the time and the maturation of mouse oocytes meiosis.All of the results have shown that the SLK is very important for the mouse oocyte meiosis. Therefore, the research of SLK is also very necessary. The article carried out a detailed study for the function of SLK in mouse oocyte meiosis.