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J. Biol. Chem., Vol. 275, Issue 44, 34407-34414, November 3, 2000
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From the Departments of Given the roles proposed for gap junctional
intercellular communication in neuronal differentiation and
growth control, we examined the effects of connexin43 (Cx43) expression
in a neuroblastoma cell line. A vesicular stomatitis virus G protein
(VSVG)-pseudotyped retrovector was engineered to co-express the green
fluorescent protein (GFP) and Cx43 in the communication-deficient
neuro-2a (N2a) cell line. The 293 GPG packaging cell line was
used to produce VSVG-pseudotyped retrovectors coding for GFP, Cx43, or
chimeric Cx43·GFP fusion protein. The titer of viral supernatant, as
measured by flow cytometry for GFP fluorescence, was approximately
2.0 × 107 colony form units (CFU)/ml and was
free of replication-competent retroviruses. After a 7-day treatment
with retinoic acid (20 µM), N2a transformants (N2a-Cx43
and N2a-Cx43·GFP) maintained the expression of Cx43 and Cx43·GFP.
Expression of both constructs resulted in functional coupling, as
evidenced by electrophysiological and dye-injection analysis.
Suppression of cell growth correlated with expression of both Cx43 or
Cx43·GFP and retinoic acid treatment. Based on morphology and
immunocytochemistry for neurofilament, no difference was observed in
the differentiation of N2a cells compared with cells expressing Cx43
constructs. In conclusion, constitutive expression of Cx43 in N2a cells
does not alter retinoic acid-induced neuronal differentiation but does
enhance growth inhibition.
Neuronal Differentiation and Growth Control of Neuro-2a Cells
After Retroviral Gene Delivery of Connexin43*
,,, and
Anatomy and Cell Biology and
§ Medical Biophysics, Child Health Research Institute,
University of Western Ontario, London, Ontario, Canada N6A 5C1 and the
¶ McGill Centre for Translational Research in Cancer, Lady
Davis Institute, McGill University, Montreal, QC, Canada H3T 1E2
*
This work was supported by the Medical Research Council of
Canada.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed: Dept. of Anatomy
and Cell Biology, Medical Sciences Building, University of Western Ontario, London, ON, Canada N6A 5C1. Tel.: 519-661-4067; Fax: 519-661-3936; E-mail: cnaus@julian.uwo.ca.
Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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