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Targeted cell‐ablation in Xenopus embryos using the conditional, toxic viral protein M2(H37A)
Author(s) -
Smith Stuart J.,
Kotecha Surendra,
Towers Norma,
Mohun Timothy J.
Publication year - 2007
Publication title -
developmental dynamics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.634
H-Index - 141
eISSN - 1097-0177
pISSN - 1058-8388
DOI - 10.1002/dvdy.21233
Subject(s) - biology , xenopus , embryo , cell fate determination , blastula , microbiology and biotechnology , programmed cell death , blastomere , embryonic stem cell , cell , wnt signaling pathway , embryogenesis , gene , genetics , apoptosis , gastrulation , transcription factor , signal transduction
Abstract Harnessing toxic proteins to destroy selective cells in an embryo is an attractive method for exploring details of cell fate and cell–cell interdependency. However, no existing “suicide gene” system has proved suitable for aquatic vertebrates. We use the M2(H37A) toxic ion channel of the influenza‐A virus to induce cell‐ablations in Xenopus laevis . M2(H37A) RNA injected into blastomeres of early stage embryos causes death of their progeny by late‐blastula stages. Moreover, M2(H37A) toxicity can be controlled using the M2 inhibitor rimantadine. We have tested the ablation system using transgenesis to target M2(H37A) expression to selected cells in the embryo. Using the myocardial MLC2 promoter, M2(H37A)‐mediated cell death causes dramatic loss of cardiac structure and function by stage 39. With the LURP1 promoter, we induce cell‐ablations of macrophages. These experiments demonstrate the effectiveness of M2(H37A)‐ablation in Xenopus and its utility in monitoring the progression of developmental abnormalities during targeted cell death experiments. Developmental Dynamics 236:2159–2171, 2007. © 2007 Wiley‐Liss, Inc.