Expression of luciferase in selected organs following delivery of naked and formulated DNA to rainbow trout (Oncorhynchus mykiss) by different routes of administration.
In the present work, the expression of luciferase in selected organs following administration of DNA delivered as naked, liposome-formulated or chitosan-formulated by different routes of administration (intramuscular, intraperitoneal and intravenous injection, immersion and anal intubation) was studied in rainbow trout (Oncorhynchus mykiss). The different formulations and routes of administration both influenced in which organs luciferase was expressed and the magnitude of expression. The highest expression levels of luciferase in the head kidney and liver were found after an intraperitoneal injection of lipoplex 2. In the spleen, the highest levels were detected after injection of naked DNA (intraperitonal or intramuscular) and lipoplex 2 (intraperitoneal). Following intravenous injection, naked DNA gave higher expression levels in the organs than the formulated plasmids and immersion and anal intubation were not effective routes of delivery as no expression of luciferase could be detected in any of the organs tested. Additionally, PCR using a primer specific for a 600 bp region of the luciferase gene pcDNA3-luc was used to assess the distribution of the plasmid itself after intramuscular and intraperitoneal injection. Positive amplification was obtained in spleen, head kidney, liver and muscle at the injection site following injection of formulated plasmids, while only muscle tissue from the injection site was positive when naked DNA was used.
Romøren
Kristine
K
Department of Pharmaceutics, School of Pharmacy, University of Oslo, Blindern P.O. Box 1068, Oslo 0316, Norway. kristine.romoren@farmasi.uio.no
Thu
Beate J
BJ
Evensen
Øystein
Ø
D000277:Adjuvants, Pharmaceutic / Q000378:metabolism; D000818:Animals; D002686:Chitin / Q000031:analogs & derivatives* / Q000276:immunology; D048271:Chitosan; D017931:DNA Primers; D007101:Immersion; D007273:Injections, Intramuscular; D007274:Injections, Intraperitoneal; D007441:Intubation, Gastrointestinal; D008081:Liposomes; D008156:Luciferases / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D017686:Oncorhynchus mykiss / Q000276:immunology*; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D014018:Tissue Distribution; D019444:Vaccines, DNA / Q000008:administration & dosage* / Q000276:immunology* / Q000378:metabolism
D000277:Adjuvants, Pharmaceutic; D017931:DNA Primers; D008081:Liposomes; D019444:Vaccines, DNA; D002686:Chitin; D048271:Chitosan; D008156:Luciferases