Gene Transfer of Prostaglandin Synthase Maintains Patency of the Newborn Lamb Arterial Duct
Autor: | Yasufumi Kaneda, Hamid Massaeli, Syed H.E. Zaidi, Tilman Humpl, James Y. Coe, Lee N. Benson, Jennifer L. Russell, Marlene Rabinovitch |
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Rok vydání: | 2005 |
Předmět: |
animal diseases
Blotting Western Prostaglandin Gene transfer Arterial duct Transfection Dinoprostone Andrology chemistry.chemical_compound parasitic diseases medicine Animals DNA Primers Sheep Base Sequence ATP synthase biology Genetic transfer Ductus Arteriosus Anatomy respiratory system medicine.anatomical_structure Animals Newborn chemistry Echocardiography Prostaglandin-Endoperoxide Synthases Mutation Pediatrics Perinatology and Child Health Circulatory system biology.protein lipids (amino acids peptides and proteins) Blood vessel Artery |
Zdroj: | Pediatric Research. 58:976-980 |
ISSN: | 1530-0447 0031-3998 |
Popis: | In congenital heart disease with left- or right-sided obstruction, prostaglandin E (PGE)1 or PGE2 is infused to maintain ductus arteriosus (DA) patency. We hypothesized that transfection of the DA with PGE synthase would lead to a greater production of PGE2 in situ and, hence, patency of the DA. The cDNA for human prostaglandin synthase was sequenced and ligated into a eukaryotic expression vector. The negative control was created by ligating the cDNA encoding the bacterial protein chloramphenicol acetyltransferase into the same plasmid. Transfection (600 microg DNA) was achieved in lambs within the first 24 h of life using the hemagglutinating virus of Japan (HVJ)-liposome transfection method with a custom-made, basket-weave-perforated catheter. Echocardiography was performed to assess DA patency until the time of sacrifice. To confirm expression of the transgene, PGE2 concentration was measured in organ culture of the DA by immunoassay and by Western immunoblotting of homogenized DA tissue. Patency of the DA was demonstrated by color Doppler in all the lambs (7/7) in which the PGE synthase was delivered, whereas functional closure was seen in the control group (6/6). The PGE2 concentration in the culture medium of the explanted DA in the treatment group was 3-fold higher than that of the control groups. Western immunoblotting confirmed the presence of PGE synthase in the treatment group. Gene transfer of PGE synthase to the DA is feasible and will maintain patency for at least 1 wk. |
Databáze: | OpenAIRE |
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