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Isolation of Distinct Cell Populations from the Developing Cerebellum by Microdissection
Published on: September 21, 2014
Impaired neurite outgrowth of src-minus cerebellar neurons on the cell adhesion molecule L1
M A Ignelzi1, D R Miller, P Soriano
1Department of Biochemistry and Biophysics, University of North Carolina School of Medicine, Chapel Hill 27599-7260.
Abstract:
The nonreceptor tyrosine protein kinases pp60c-src, p59fyn, and pp62c-yes are localized in growth cones of developing neurons, but their function is undefined. To determine whether these tyrosine kinases were capable of regulating substrate-dependent axon growth, cultures of cerebellar neurons from wild-type, src-, fyn-, and yes- mice were analyzed for neurite outgrowth on the neural cell adhesion molecule L1 or the extracellular matrix protein laminin. The rate of neurite extension on L1 was reduced in src-, but not in fyn- or yes- neurons. Neurite extension on laminin was unaltered in src-, fyn-, or yes- neurons, indicating that pp60c-src, p59fyn, or pp62c-yes is not likely to participate in integrin-dependent axon growth. These results demonstrate that pp60c-src is a component of the intracellular signaling pathway in L1-mediated axonal growth and suggest that Src-related nonreceptor tyrosine kinases may have distinct, nonredundant functions in the nervous system.
Insights
The study found that the tyrosine kinase pp60c-src is crucial for axon growth mediated by the L1 molecule in developing neurons. Other related kinases, fyn and yes, do not play this role, suggesting distinct functions in the nervous system.
Area of Science:
- Neuroscience
- Cell Biology
- Molecular Biology
Background:
- Nonreceptor tyrosine protein kinases, including pp60c-src, p59fyn, and pp62c-yes, are present in neuronal growth cones.
- The specific roles of these kinases in neuronal development and axon growth remain largely unknown.
Purpose of the Study:
- To investigate the function of Src-related nonreceptor tyrosine kinases in substrate-dependent axon growth.
- To determine if pp60c-src, p59fyn, and pp62c-yes regulate neurite outgrowth on specific extracellular molecules.
Main Methods:
- Cerebellar neurons from wild-type, src-, fyn-, and yes- knockout mice were cultured.
- Neurite outgrowth was analyzed on the neural cell adhesion molecule L1 and the extracellular matrix protein laminin.
Main Results:
- Neurite extension on L1 was significantly reduced in src- deficient neurons, while fyn- and yes- deficient neurons showed no difference.
- Neurite extension on laminin was not affected in neurons lacking src, fyn, or yes.
- These findings indicate pp60c-src is involved in L1-mediated axon growth but not integrin-dependent growth on laminin.
Conclusions:
- pp60c-src is a key component of the intracellular signaling pathway for L1-mediated axonal growth.
- Src-related nonreceptor tyrosine kinases likely possess distinct and non-redundant functions within the nervous system.
