J Biomed Res.  2013 Jun;14(2):60-64.

Activation of small Rho GTPases by blebbistatin in PC12 cells

Affiliations
  • 1Department of Biochemistry, College of Medicine and Medical Research Institute, Chungbuk National University, Cheongju 361-763, Korea. eyshin@chungbuk.ac.kr

Abstract

Neuronal differentiation is a complex biological process accompanying cytoskeletal reorganization, including neurite outgrowth and growth cone formation. Therefore, neuronal differentiation is critically regulated by actin-related signaling proteins, such as small Rho GTPases, guanine nucleotide exchange factors (GEFs), and myosins. This study will demonstrate the change in activity of three small Rho GTPases, Rac, Cdc42, and Rho A, by treatment with blebbistatin (BBS), a specific inhibitor for myosin, during bFGF-induced neurite outgrowth in PC12 cells. Treatment with BBS induced morphological changes in growth cones and neurites during differentiation. A marked increase in protrusion and filopodia structures in growth cones, the shaft of neuritis, and cell membranes was observed in the cells treated with BBS. Activity of Rho GTPases showed the alterations in response to BBS. Activities of both Rac and Rho A were inhibited by BBS in a time-dependent manner. By contrast, Cdc42 activity was not changed by BBS. These results suggest that inactivation of myosin II by BBS induced morphological changes in neurites and growth cones and distinct regulation of three Rho GTPases during differentiation of PC12 cells.

Keyword

Rho GTPase; Rac; Rho A; Cdc42; blebbistatin

MeSH Terms

Animals
Biological Processes
Cell Membrane
Growth Cones
Guanine Nucleotide Exchange Factors
Heterocyclic Compounds with 4 or More Rings
Myosin Type II
Myosins
Neurites
Neuritis
Neurons
PC12 Cells*
Proteins
Pseudopodia
rho GTP-Binding Proteins*
Guanine Nucleotide Exchange Factors
Heterocyclic Compounds with 4 or More Rings
Myosin Type II
Myosins
Proteins
rho GTP-Binding Proteins
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