In:
The Journal of Neuroscience, Society for Neuroscience, Vol. 21, No. 14 ( 2001-07-15), p. 5191-5202
Abstract:
Neuronal differentiation involves Rac and Cdc42 GTPases. α-Chimaerin, a Rac/Cdc42 regulator, occurs as α1- and alternatively spliced Src homology 2 (SH2) domain-containing α2-isoforms. α2-chimaerin mRNA was highly expressed in the rat embryonic nervous system, especially in early postmitotic neurons. α1-chimaerin mRNA was undetectable before embryonic day 16.5. Adult α2-chimaerin mRNA was restricted to neurons within specific brain regions, with highest expression in the entorhinal cortex. α2-chimaerin protein localized to neuronal perikarya, dendrites, and axons. The overall pattern of α2-chimaerin mRNA expression resembles that of cyclin-dependent kinase regulator p35 (CDK5/p35) which participates in neuronal differentiation and with which chimaerin interacts. To determine whether α2-chimaerin may have a role in neuronal differentiation and the relevance of the SH2 domain, the morphological effects of both chimaerin isoforms were investigated in N1E-115 neuroblastoma cells. When plated on poly-lysine, transient α2-chimaerin but not α1-chimaerin transfectants formed neurites. Permanent α2-chimaerin transfectants generated neurites whether or not they were stimulated by serum starvation, and many cells were enlarged. Permanent α1-chimaerin transfectants displayed numerous microspikes and contained F-actin clusters, a Cdc42-phenotype, but generated few neurites. In neuroblastoma cells, α2-chimaerin was predominantly soluble with some being membrane-associated, whereas α1-chimaerin was absent from the cytosol, being membrane- and cytoskeleton-associated, paralleling their subcellular distribution in brain. Transient transfection with α2-chimaerin mutated in the SH2 domain (N94H) generated an α1-chimaerin-like phenotype, protein partitioned in the particulate fraction, and in NGF-stimulated pheochromocytoma cell line 12 (PC12) cells, neurite formation was inhibited. These results indicate a role for α2-chimaerin in morphological differentiation for which its SH2 domain is vital.
Type of Medium:
Online Resource
ISSN:
0270-6474
,
1529-2401
DOI:
10.1523/JNEUROSCI.21-14-05191.2001
Language:
English
Publisher:
Society for Neuroscience
Publication Date:
2001
detail.hit.zdb_id:
1475274-8
SSG:
12
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