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p160 RhoA-binding kinase ROKalpha induces neurite retraction
1Department of Physiological Chemistry, Faculty of Pharmaceutical Sciences, Kyoto University, Sakyo-ku, Kyoto 606, Japan.
The Journal of Biological Chemistry
|February 28, 1998
Summary
p160 RhoA-binding kinase ROKalpha induces neurite retraction downstream of Rho in neuronal cells. This finding identifies a key effector in prostaglandin E receptor signaling pathways.
Area of Science:
- Cell Biology
- Neuroscience
- Molecular Biology
Background:
- Prostaglandin E receptor EP3 activation causes neurite retraction via Rho.
- The specific downstream effector of Rho in this process remained unidentified.
Purpose of the Study:
- To investigate the role of p160 RhoA-binding kinase ROKalpha as a downstream effector of Rho in neurite retraction.
- To examine the morphological effects of ROKalpha on nerve growth factor-differentiated PC12 cells.
Main Methods:
- Microinjection of ROKalpha catalytic domain and kinase-deficient mutant into PC12 cells.
- Pre-injection with C3 exoenzyme to inactivate Rho.
- Microinjection of ROKalpha domains to inhibit EP3 receptor-induced neurite retraction.
Main Results:
- Catalytic domain of ROKalpha rapidly induced neurite retraction, similar to constitutively active RhoV14.
- Kinase-deficient ROKalpha did not induce neurite retraction.
- ROKalpha inhibition blocked EP3 receptor-induced neurite retraction, even when Rho was inactivated.
Conclusions:
- ROKalpha acts downstream of Rho to induce neurite retraction in neuronal cells.
- ROKalpha is a key effector in the EP3 receptor signaling pathway regulating neuronal morphology.