Format

Send to:

Choose Destination
See comment in PubMed Commons below
Dev Biol. 2000 Feb 15;218(2):199-205.

Vaccinia as a tool for functional analysis in regenerating limbs: ectopic expression of Shh.

Author information

  • 1Developmental Biology Center and Department of Developmental and Cell Biology, University of California at Irvine, Irvine, California 92697, USA.

Abstract

Axolotls, with their extensive abilities to regenerate as adults, provide a useful model in which to study the mechanisms of regeneration in a vertebrate, in hopes of understanding why other vertebrates cannot regenerate. Although the expression of many genes has been described in regeneration, techniques for functional analysis have so far been limited. In this paper we demonstrate a new method for efficient overexpression of foreign genes in axolotls. Using vaccinia virus expressing beta-galactosidase microinjected into regenerating limbs, we show that vaccinia can infect both dividing and nondividing limb cells. The site of infection remains discrete and there is no secondary spread of infection to nearby cells. beta-Gal is expressed at high levels in blastema cells for about a week and in differentiated cells for longer. Blastemas that have been injected with vaccinia at different stages regenerate normally. As a test of the utility of vaccinia for functional analysis in regeneration, we constructed a virus expressing Shh and injected it into the anterior of regenerating limbs. Ectopic Shh expression caused extra digits, carpals, and tarsals in the hands and feet of regenerating limbs, suggesting that despite differences in the timing of expression and the eventual pattern, the function of Shh appears to be similar to that in the developing limbs of other vertebrates. Our results demonstrate that vaccinia virus is an excellent vector for ectopically expressing genes for secreted proteins and is a useful tool to study the function of signaling molecules during the process of regeneration in urodeles.

Copyright 2000 Academic Press.

PMID:
10656763
[PubMed - indexed for MEDLINE]
Free full text
PubMed Commons home

PubMed Commons

0 comments
How to join PubMed Commons

    Supplemental Content

    Full text links

    Icon for Elsevier Science
    Loading ...
    Write to the Help Desk