The role of protein kinase B (PKB) in the regulation of early development of fertilized eggs in mouse and its biological function on the development of mouse 1\|cell stage fertilized eggs were studied.The kinase activity of PKB was detected,and mRNA and protein level of PKB both in MⅡ oocytes and 1\|cell stage fertilized eggs were semi\|quantitatively assayed by Western blotting and RT\|PCR.10 μmol/L LY294002,the inhibitor of PKB,was microinjected into 1\|cell stage fertilized eggs of mouse to observe its effect on the development of mouse 1\|cell stage fertilized eggs.The results showed that the activity of PKB increased obviously after the fertilization,the mRNA and protein level of PKB also increased obviously in 1\|cell stage fertilized eggs,while in MⅡ oocytes the mRNA level was not detected.The protein level was lower than that in the 1\|cell stage fertilized eggs.After microinjecting LY294002 the development from 1\|cell stage to 2\|cell stage was inhibited obviously.All the results demonstrate that PKB exists in the fertilized eggs,and the activity,expression and transcription level increase after fertilization.PKB may play a positive role during the early development of fertilized eggs in mouse from 1\|cell to 2\|cell stage,but the mechanism is still unknown.These experiments are helpful for the further study of PKB on the regulation of early development of fertilized eggs.
Protein kinase C (PKC) is a family of serine/threonine protein kinases, and its multiple isoforms are expressed in various mammalian tissues. The expressions of PKC α and PKC δ at protein and mRNA level in mouse testis were identified by Western blotting and RT\|PCR. The expression of both PKC isoenzymes in the developing mouse testis was also examined. In testes of mouse at various developmental stages, both the protein and the mRNA of PKC\|α were uniformity; but the PKC\|δ expression occurred in the testes of 3\|week old, perhaps even relatively late in spermatid development. The results suggest that each isoenzyme may have different roles in processing and modulating physiological cellular responses of spermatogenesis.
