Differential expression of protein kinases in cultured primary neurons derived from the cerebral cortex, hippocampus, and sympathetic ganglia

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Abstract
Protein kinases play pivotal roles in the development of the nervous system. They are involved in almost every stage of neuronal development, from initial proliferation and differentiation of progenitor cells to pathfinding of neurities and formation of synapses. Activation of protein kinases is also critical for neuronal cell survival. To gain further insights into kinases in neural development, we studied the expression patterns of protein kinases in three cultured primary neurons by degenerate primer-based reverse transcription-polymerase chain reaction (PCR) and DNA sequencing, taking advantage of all known kinases containing a conserved catalytic domain. Our data demonstrated that the expression patterns of kinases in various cultured neurons are not only different from those of non-neural tissues, but also distinct among neurons derived from discrete origins. For example, FGF receptor 1 is predominantly expressed in hippocampal neurons. As this approach may be biased during PCR and cloning steps, an RNase protection assay was employed to verify the expression levels of six kinases in cultured neurons. Results from the RNase protection assay did generally confirm those obtained by the PCR-based method. However, quantitative nature of the latter was dependent on numbers of clones analyzed, and discrepancy of expression levels of kinases detected by the two methods was sometimes observed.