In vivo plasmid DNA electroporation resulted in transfection of satellite cells and lasting transgene expression in regenerated muscle fibers.
In vivo plasmid DNA electroporation resulted in elevated and lasting transgene expression in skeletal muscles. But the nature of the cells that contributed to sustained gene expression remains unknown. We followed the fate of plasmid DNA delivered with electroporation and systematically investigated the time course and location of transgene expression in muscle tissues both with GFP and luciferase. Furthermore, satellite cell activation after electroporation was confirmed by RT-PCR and immunohistochemistry analysis. The activated satellite cells were shown to be able to uptake the injected plasmid DNA and express transgene products as regenerated myocytes. We found that cells with longer gene expression durations were mostly regenerated muscle fibers. In contrast, expression in pre-existing muscle fibers was rather transient. We also presented in this study that immune response to transgene products might hamper the lasting gene expression. Based on these observations, we proposed that the underlying mechanism for prolonged transgene expression in the muscles after electroporation is related to the activation and transfection of myogenic satellite cells which subsequently developed into regenerated muscle fibers.
Peng
Baowei
B
School of Life Sciences and Biotechnology, Shanghai Jiaotong University, Shanghai, PR China.
Zhao
Yonggang
Y
Lu
Huili
H
Pang
Wengka
W
Xu
Yuhong
Y
D000818:Animals; D015496:CD4-Positive T-Lymphocytes / Q000378:metabolism; D018414:CD8-Positive T-Lymphocytes / Q000378:metabolism; D004247:DNA / Q000235:genetics*; D018274:Electroporation / Q000379:methods*; D005260:Female; D015870:Gene Expression / Q000235:genetics*; D051379:Mice; D018485:Muscle Fibers, Skeletal / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics*; D051381:Rats; D012038:Regeneration; D032496:Satellite Cells, Skeletal Muscle / Q000378:metabolism*; D014162:Transfection; D019076:Transgenes / Q000235:genetics*
D004247:DNA