In vivo transfection and expression of exogenous genes within proximal tubules of rats.
An in vivo technique for expressing exogenous DNA within the renal cortical proximal tubules of rats was was developed and validated. Cationic liposomes were complexed with either water or 25 microg of plasmid DNA containing the lac Z gene, and injected into the left renal artery of anesthetized rats. Under transcriptional control of the chicken beta-actin, SV40, and CMV promoters, beta-galactosidase expression was consistently observed in rats that received plasmid DNA, while no blue staining was observed in kidneys of control rats. Using an injection volume of 400 microl, lac Z expression was confined to the proximal tubules, with no lac Z expression noted within serial sections of the liver, adrenal, lung, spleen, or heart. Because lac Z expression was observed for up to 20 days without apparent systemic or renal insult, this technique may be a valuable tool for evaluating physiological and pathophysiological changes associated with gene expression within this isolated site.
Marley
W S
WS
Department of Physiology, Colorado State University, Fort Collins 80523, USA.
Wilke
L
L
Bowen
R A
RA
D000199:Actins / Q000235:genetics; D000818:Animals; D003587:Cytomegalovirus / Q000235:genetics; D015870:Gene Expression; D017930:Genes, Reporter; D007687:Kidney Tubules, Proximal / Q000378:metabolism*; D007763:Lac Operon; D008297:Male; D011401:Promoter Regions, Genetic; D051381:Rats; D017207:Rats, Sprague-Dawley; D013539:Simian virus 40 / Q000235:genetics; D014162:Transfection; D001616:beta-Galactosidase / Q000235:genetics
D000199:Actins; D001616:beta-Galactosidase