Transduction of bovine adrenal chromaffin cells using a recombinant adenovirus expressing GFP.
Insertion of genetic material into bovine chromaffin cells employing various techniques have produced low to moderate transduction rates. Recent technology using adenoviral gene transfer has become one of the most powerful methods for introducing genes into mammalian cells. We examined whether a recombinant adenovirus could provide a convenient vector to transfer genes of interest for mechanistic studies on chromaffin cells. Our results show that 100% transduction of chromaffin cells was accomplished within 18 h with a recombinant adenovirus as revealed by the expression of green fluorescent protein (GFP) in chromaffin cells. Transduction was dependent on time and viral titer but independent of cell age and density in culture. There was no effect of the recombinant adenovirus on the secretory function [3H]-norepinephrine ([3H]-NE release) of the chromaffin cells. The results demonstrate that the recombinant adenovirus provides an effective process for the complete transfection of bovine chromaffin cells with a selected gene.
Li
Xinying
X
Department of Pharmacology, University of Nebraska Medical Center, 986260 Nebraska Medical Center, Omaha, NE 68198-6260, USA.
Drakulich
Desinee A
DA
Zhang
Peijin
P
Shen
Meilan
M
Weber
Gregory A
GA
Ikezu
Tsuneya
T
Hexum
Terry D
TD
D000256:Adenoviridae / Q000235:genetics*; D000313:Adrenal Medulla / Q000378:metabolism; D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D019439:Chromaffin Cells / Q000378:metabolism*; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000378:metabolism*; D009638:Norepinephrine / Q000378:metabolism; D011995:Recombination, Genetic; D014161:Transduction, Genetic / Q000379:methods*; D014162:Transfection / Q000379:methods
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D009638:Norepinephrine