Tubulin beta 4B chain is involved in the cell cycle process of mouse spermatogonia through CDK2

Abstract Tubb4b (tubulin beta-4b chain) is essential for spermatogonia cell growth and development as a microtubule network protein, and its regulatory mechanism remains to be studied. To deeply understand the role of Tubb4b in spermatogonia cell proliferation and the cell cycle, we studied the role of spermatogonia in the control and TUBB4B-KO groups in different cell cycle phases by flow cytometry. We detected the crucial protein CDK2 in each stage by qRT–PCR and Western blotting. The results showed that under the induction of serum starvation, thymidine, and nocodazole under different conditions, mouse spermatogonia remained in the G0-G1 phase, S phase, and G2-M phase, respectively, and the change in the spermatogonia cycle in the Tubb4b-KO group was slower than that in the control group. In the study of cycle synchronization, it was found that the mRNA level of Cdk2 in the G2-M phase increased significantly (P < 0.001), accompanied by the accumulation of CDK2 protein. In the Tubb4b-KO group, the mRNA level of Cdk2 increased significantly in the G0-G1 phase (P < 0.05), decreased significantly in the S phase (P < 0.05) and was accompanied by the fluctuation of CDK2 protein. These data suggest that Tubb4b has the potential to play an important role during the spermatogonial cell cycle by affecting the fluctuating expression of CDK2.