Cdk5 regulates axonal transport and phosphorylation of neurofilaments in cultured neurons.
Phosphorylation has long been considered to regulate neurofilament (NF) interaction and axonal transport, and, in turn, to influence axonal stability and their maturation to large-caliber axons. Cdk5, a serine/threonine kinase homologous to the mitotic cyclin-dependent kinases, phosphorylates NF subunits in intact cells. In this study, we used two different haptenized NF subunits and manipulated cdk5 activity by microinjection, transfection and pharmacological inhibition to monitor the effect of Cdk5-p35 on NF dynamics and transport. We demonstrate that overexpression of cdk5 increases NF phosphorylation and inhibits NF axonal transport, whereas inhibition both reduces NF phosphorylation and enhances NF axonal transport in cultured chicken dorsal-root-ganglion neurons. Large phosphorylated-NF 'bundles' were prominent in perikarya following cdk5 overexpression. These findings suggest that Cdk5-p35 activity regulates normal NF distribution and that overexpression of Cdk5-p35 induces perikaryal accumulation of phosphorylated-NFs similar to those observed under pathological conditions.
Shea
Thomas B
TB
Center for Cellular Neurobiology and Neurodegeneration Research, Department of Biological Sciences, University of Massachusetts, Lowell, One University Avenue, Lowell, MA 01854, USA. thomas_shea@uml.edu
Yabe
Jason T
JT
Ortiz
Daniela
D
Pimenta
Aurea
A
Loomis
Patti
P
Goldman
Robert D
RD
Amin
Niranjana
N
Pant
Harish C
HC
D000818:Animals; D001369:Axons / Q000378:metabolism*; D001693:Biological Transport, Active / Q000502:physiology; D002478:Cells, Cultured; D002642:Chick Embryo; D051360:Cyclin-Dependent Kinase 5; D018844:Cyclin-Dependent Kinases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D007382:Intermediate Filaments / Q000378:metabolism*; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D016900:Neurofilament Proteins / Q000302:isolation & purification / Q000378:metabolism*; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D010766:Phosphorylation; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014162:Transfection
D009419:Nerve Tissue Proteins; D016900:Neurofilament Proteins; D011994:Recombinant Proteins; C089495:neuronal Cdk5 activator (p25-p35); D051360:Cyclin-Dependent Kinase 5; D018844:Cyclin-Dependent Kinases