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Using Chemical Inhibitors to Study the Role of the Cyclin Dependent Kinase 5 (CDK5) Protein in the Nerve Growth Factor Induced Neuronal Differentiation of PC12 Cells
Author(s) -
Perez Elizabeth,
Aguanno Ann
Publication year - 2006
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.20.5.a969-c
Subject(s) - cyclin dependent kinase 5 , nerve growth factor , microbiology and biotechnology , biology , morphogenesis , cellular differentiation , phenotype , kinase , growth factor , cyclin dependent kinase , protein kinase a , cyclin dependent kinase 2 , cell cycle , cell , genetics , receptor , gene
Cyclin‐dependent kinase 5 (CDK5) is classified as a member of the cyclin‐dependent kinase family based on sequence homology. Unlike most members of this family, however, CDK5 does not participate in the progression of the cell cycle. CDK5, although ubiquitously expressed, exhibits an activity profile restricted to the nervous system due to the neuron‐specific location of its activating partners, p35 and p39. This data, coupled with its known substrates (e.g. Tau), has strongly suggested that CDK5 is essential for neuronal migration, morphogenesis and survival. PC12 cells are a widely used model for the neuronal differentiation process due to their elaboration of a neuronal‐like phenotype in response to a number of growth factors (e.g. Nerve Growth Factor (NGF), Fibroblast Growth Factor). Here we report an alteration in the NGF‐induced neuronal differentiation process of PC12 cells through the chemical inhibition of CDK5 with either olomoucine and 6‐(γ, γ, dimethylallyloamino)purine. Specifically, we present a simple but highly reproducible assay for quantifying the morphology of the growth factor‐induced neuronal differentiation of these cells and any alterations in this phenotype that result from the inhibition of CDK5. Our data shows that specific, morphological attributes, easily identified via light microscopy, can be quantified and subjected to rigorous statistical testing using this approach. Moreover, our data demonstrates that CDK5 plays a role in the establishment of the NGF‐induced neuronal phenotype in PC12 cells and further reveals when CDK5 participates in this multi‐step, growth factor‐induced differentiation process.