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The Changes of Biological Characters and Podocytes Associated Proteins Distribution Profile in Undifferentiated and Differentiated Podocytes
Hua-Jun Jiang, Chun Zhang*, Ying Chang1, Zhong-Hua Zhu, Jian-She Liu, An-Guo Deng
Department of Nephrology, Union Hospital, Tongji Medical College, Huazhong University of Science & Technology, Wuhan 430022, China; 1Institute of Liver Disease, Tongji Hospital, Tongji Medical College, Huazhong University of Science & Technolog
Abstract: Conditionally immortalized murine podocyte cell line， which was established from transgenic H-2Kb-tsA58 mice， was cultured in RPMI 1640 medium at 33 ℃ permissive conditions and shifted to 37 ℃ non-permissive conditions to induce differentiation. Podocytes morphology changes， growth curve， proliferation ability and cell cycle were measured respectively in undifferentiated and differentiated podocytes. The expressions of CD2AP， α-actinin， nephrin and cytoskeletal proteins (i.e. F-actin and tubulin) were detected by laser scanning confocal microscope. The results demonstrated that in differentiated podocytes， cell morphology changed along with secondary foot process formation and reproductive activity descended. Cells in G0/G1 period accumulated and in S， G2/M period decreased. The expressions of CD2AP， nephrin and α-actinin were elevated in differentiated podocytes. The distribution profiles of podocyte associated proteins and cytoskeletal proteins apparently altered. In conclusion， differentiated podocytes biological characters changed and cytoskeleton rearranged. CD2AP， nephrin， α-actinin， F-actin and tubulin may play a crucial role in podocytes differentiation.
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