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Chromatin Immunoprecipitation from Dorsal Root Ganglia Tissue following Axonal Injury
Published on: July 20, 2011
Dynamic changes of p27(kip1) and Skp2 expression in injured rat sciatic nerve
Ai-Guo Shen1, Shu-Xian Shi, Meng-Ling Chen
1The Jiangsu Province Key Laboratory of Neuroregeneration, Nantong University, Nantong, P.R. China.
Abstract:
S phase kinase-associated protein 2 (Skp2), an F-box protein, is required for the ubiquitination and consequent degradation of p27(kip1). Previous reports have showed that p27(kip1 )played important roles in cell cycle regulation and neurogenesis in the developing central nervous system. But the distribution and function of p27(kip1 )and Skp2 in nervous system lesion and regeneration remains unclear. In this study, we observed that they were expressed mainly in both Schwann cells and axons in adult rat sciatic nerve. Sciatic nerve crush and transection resulted in a significant up-regulation of Skp2 and a down-regulation of p27(kip1). By immunochemistry, we found that in the distal stumps of transected nerve from the end to the edge, the appearance of Skp2 in the edge is coincided with the decrease in p27(kip1) levels. Changes of them were inversely correlated. Results obtained by coimmunoprecipitation and double labeling further showed their interaction in the regenerating process. Thus, these results indicate that p27(kip1 )and Skp2 likely play an important role in peripheral nerve injury and regeneration.
Insights
Peripheral nerve injury involves S phase kinase-associated protein 2 (Skp2) and p27(kip1). Skp2 levels increase while p27(kip1) levels decrease, indicating their role in nerve regeneration.
Area of Science:
- Neuroscience
- Cell Biology
Background:
- S phase kinase-associated protein 2 (Skp2) targets p27(kip1) for degradation.
- p27(kip1) is crucial for cell cycle regulation and neurogenesis.
- The roles of Skp2 and p27(kip1) in peripheral nerve injury and regeneration are not well understood.
Purpose of the Study:
- To investigate the distribution and function of p27(kip1) and Skp2 in adult rat sciatic nerve following injury.
- To elucidate the relationship between Skp2 and p27(kip1) during peripheral nerve regeneration.
Main Methods:
- Sciatic nerve crush and transection models in adult rats.
- Immunohistochemistry to detect protein expression and localization.
- Co-immunoprecipitation and double labeling to assess protein interactions.
Main Results:
- Skp2 and p27(kip1) are expressed in Schwann cells and axons of the adult rat sciatic nerve.
- Nerve injury (crush and transection) led to increased Skp2 and decreased p27(kip1) expression.
- Skp2 upregulation inversely correlated with p27(kip1) downregulation in regenerating nerve stumps.
- Skp2 and p27(kip1) interact during the nerve regeneration process.
Conclusions:
- Skp2 and p27(kip1) are dynamically regulated following peripheral nerve injury.
- Their inverse correlation and interaction suggest a significant role in peripheral nerve regeneration.
