Transfection genique directe dans le rein de rat in vivo

Translated title of the contribution: Direct gene transfer in the rat kidney in vivo

D. Stephan, B. Gasser, H. San, M. Schubnel, G. J. Nabel, E. G. Nabel

Research output: Contribution to journalArticle

Abstract

Gene delivery to the kidney has both experimental and therapeutic potential in hypertension, although the delivery methods, distribution of transgene and subsequent inflammatory response, have been poorly characterized. In adult male Sprague-Dawley rats (200 g, n = 26), the left iliac artery was catheterized and a small catheter (Microbore Tygon S-54-HL) was advanced to the origin of the left renal artery. Loops were tied transiently around the aorta and below the renal arterial bifurcation. After flushing the kidney, the renal vein was tied and 500 μL of transfection solution was instilled. After 15 min all the loops were released, the catheter was removed and the left iliac artery ligated. Both replication- defective adenovirus (ADV) constructions used were based on an Ad5 derivative with a partial E3 deletion. Virus ADV-chloramphenicol acetyl transferase (CAT) and ADV-human placental alkaline phosphatase (hpAP), 10 8, 3 x 10 8, 10 9 and 10 10 plaques forming units/mL (pfu/mL), were used respectively to compare the degree of transfection (CAT) and to localize the transgene in the kidney (hpAP), 48 h after transfection. Controls were infused with vehicle. ADV-CAT 10 10 pfu/mL induced a gene expression, respectively, 1.4 (NS), 12 (p <0.001) and 28 (p <0.001) fold greater than the 10 9, 3 x 10 8 and 10 8 pfu/mL formulations. HpAP staining was located in the juxta-medullary part of the cortex, predominantly in the interstitium. Genetically-modified cells were indentified as endothelial cells, mainly in peritubular capillaries but also in efferent arterioles and hilar arteries. Highly efficient gene transfer achieved with ADV-hpAP 10 10 pfu/mL was associated with focal necrosis of the proximal convoluted tubules. No changes were observed with the other vital concentrations. Gene delivery, mediated by a replication- defective ADV, to one rat kidney via the renal artery, induced a dose- dependent gene expression located in endothelial cells in peritubular capillaries. Toxicity was observed only with the highest vital concentration.

Original languageFrench
Pages (from-to)1127-1130
Number of pages4
JournalArchives des Maladies du Coeur et des Vaisseaux
Volume90
Issue number8
StatePublished - 1997
Externally publishedYes

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Adenoviridae
Kidney
Transfection
Human Adenoviruses
Iliac Artery
Genes
Renal Artery
Chloramphenicol
Transferases
Transgenes
Catheters
Endothelial Cells
Gene Expression
Renal Veins
Arterioles
Sprague Dawley Rats
Aorta
Necrosis
Arteries
Staining and Labeling

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine

Cite this

Stephan, D., Gasser, B., San, H., Schubnel, M., Nabel, G. J., & Nabel, E. G. (1997). Transfection genique directe dans le rein de rat in vivo. Archives des Maladies du Coeur et des Vaisseaux, 90(8), 1127-1130.

Transfection genique directe dans le rein de rat in vivo. / Stephan, D.; Gasser, B.; San, H.; Schubnel, M.; Nabel, G. J.; Nabel, E. G.

In: Archives des Maladies du Coeur et des Vaisseaux, Vol. 90, No. 8, 1997, p. 1127-1130.

Research output: Contribution to journalArticle

Stephan, D, Gasser, B, San, H, Schubnel, M, Nabel, GJ & Nabel, EG 1997, 'Transfection genique directe dans le rein de rat in vivo', Archives des Maladies du Coeur et des Vaisseaux, vol. 90, no. 8, pp. 1127-1130.
Stephan D, Gasser B, San H, Schubnel M, Nabel GJ, Nabel EG. Transfection genique directe dans le rein de rat in vivo. Archives des Maladies du Coeur et des Vaisseaux. 1997;90(8):1127-1130.
Stephan, D. ; Gasser, B. ; San, H. ; Schubnel, M. ; Nabel, G. J. ; Nabel, E. G. / Transfection genique directe dans le rein de rat in vivo. In: Archives des Maladies du Coeur et des Vaisseaux. 1997 ; Vol. 90, No. 8. pp. 1127-1130.
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