Source Paper
Rho Kinase Inhibition Enhances Axonal Regeneration in the Injured CNS
Alyson E. Fournier, Bayan T. Takizawa, Stephen M. Strittmatter
Journal of Neuroscience • 2003
Corticospinal Tract Lesion and Regeneration
Objective: To assess axonal sprouting and locomotor recovery following corticospinal tract lesions in adult rats treated with Y-27632 or C3 transferase, and to demonstrate that Rho kinase inhibition enhances axonal regeneration in the injured CNS
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Equipment1
not specified • not specified • not specified • not specified
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Protocol Steps
Rho activation measurement in vitro
Perform binding assay to measure Rho activity in PC12 and dorsal root ganglion cell lysates
Note: Detect increased levels of GTP Rho after Nogo-66 stimulation
View evidence from paper
“Using a binding assay to measure Rho activity, we detected increased levels of GTP Rho in PC12 and dorsal root ganglion (DRG) cell lysates after Nogo-66 stimulation”
Test Rho inactivation with C3 transferase in vitro
Apply C3 transferase to promote neurite outgrowth of chick DRG neurons in vitro
Note: In vitro testing of C3 transferase effectiveness
View evidence from paper
“Rho inactivation with C3 transferase promotes neurite outgrowth of chick DRG neurons in vitro”
Perform corticospinal tract lesion in adult rats
Create corticospinal tract lesions in adult rats for in vivo testing
Note: Lesion model used for both C3 transferase and Y-27632 treatment groups
View evidence from paper
“it fails to promote neurite outgrowth after corticospinal tract (CST) lesions in the adult rat”
Treat lesioned rats with C3 transferase
Administer C3 transferase to lesioned rats using the delivery method specified
Note: C3 transferase failed to promote neurite outgrowth with the delivery method used
View evidence from paper
“with the delivery method used here, it fails to promote neurite outgrowth after corticospinal tract (CST) lesions in the adult rat”
Treat lesioned rats with Y-27632
Administer Y-27632 to lesioned rats to inhibit p160ROCK and promote axonal sprouting
Note: Y-27632 treatment enhances sprouting of CST fibers in vivo
View evidence from paper
“Y-27632 enhances sprouting of CST fibers in vivo and accelerates locomotor recovery after CST lesions in adult rats”
Assess locomotor recovery
Evaluate locomotor recovery in treated and control rats following CST lesions
Note: Y-27632 treatment accelerates locomotor recovery
View evidence from paper
“Y-27632 enhances sprouting of CST fibers in vivo and accelerates locomotor recovery after CST lesions in adult rats”