Targeted cell-ablation in Xenopus embryos using the conditional, toxic viral protein M2(H37A).
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.
Smith
Stuart J
SJ
Division of Developmental Biology, National Institute for Medical Research, The Ridgeway, Mill Hill, London, United Kingdom.
Kotecha
Surendra
S
Towers
Norma
N
Mohun
Timothy J
TJ
D000818:Animals; D016923:Cell Death / Q000187:drug effects*; D004625:Embryo, Nonmammalian / Q000166:cytology* / Q000187:drug effects*; D018014:Gene Transfer Techniques; D009980:Influenza A virus / Q000737:chemistry; D007473:Ion Channels / Q000633:toxicity; D008722:Methods; D014118:Toxins, Biological / Q000235:genetics / Q000494:pharmacology*; D014764:Viral Proteins / Q000633:toxicity*; D014982:Xenopus laevis
D007473:Ion Channels; D014118:Toxins, Biological; D014764:Viral Proteins