title: Noninvasive optical diagnostics of enhanced green fluorescent protein expression in skeletal muscle for comparison of electroporation and sonoporation efficiencies.
abstract: We highlight the options available for noninvasive optical diagnostics of reporter gene expression in mouse tibialis cranialis muscle. An in vivo multispectral imaging technique combined with fluorescence spectroscopy point measurements has been used for the transcutaneous detection of enhanced green fluorescent protein (EGFP) expression, providing information on location and duration of EGFP expression and allowing quantification of EGFP expression levels. For EGFP coding plasmid (pEGFP-Nuc Vector, 10  μg/50  ml 10  μg/50  ml ) transfection, we used electroporation or ultrasound enhanced microbubble cavitation [sonoporation (SP)]. The transcutaneous EGFP fluorescence in live mice was monitored over a period of one year using the described parameters: area of EGFP positive fibers, integral intensity, and mean intensity of EGFP fluorescence. The most efficient transfection of EGFP coding plasmid was achieved, when one high voltage and four low voltage electric pulses were applied. This protocol resulted in the highest short-term and long-term EGFP expression. Other electric pulse protocols as well as SP resulted in lower fluorescence intensities of EGFP in the transfected area. We conclude that noninvasive multispectral imaging technique combined with fluorescence spectroscopy point measurements is a suitable method to estimate the dynamics and efficiency of reporter gene transfection in vivo.
authors.affiliation: Vytautas Magnus University, Biophysical Research Group, Faculty of Natural Sciences, Vileikos 8, Kaunas LT-44404, LithuaniabVytautas Magnus University, Department of Biochemistry, Faculty of Natural Sciences, Vileikos 8, Kaunas LT-44404, Lithuania.
authors.forename: Mindaugas
authors.initials: M
authors.lastname: Tamošiūnas
authors.affiliation: Institute of Electronics and Computer Science, 14 Dzerbenes Street, Riga LV-1006, Latvia.
authors.forename: Roberts
authors.initials: R
authors.lastname: Kadikis
authors.affiliation: University of Latvia, Institute of Atomic Physics and Spectroscopy, 19 Rainis Boulevard, Riga LV-1586, Latvia.
authors.forename: Inga
authors.initials: I
authors.lastname: Saknite
authors.affiliation: Lithuanian Sports University, Institute of Sports Sciences and Innovation, Sporto 6, LT-44221 Kaunas, Lithuania.
authors.forename: Juozas
authors.initials: J
authors.lastname: Baltušnikas
authors.affiliation: Lithuanian Sports University, Institute of Sports Sciences and Innovation, Sporto 6, LT-44221 Kaunas, Lithuania.
authors.forename: Audrius
authors.initials: A
authors.lastname: Kilikevičius
authors.affiliation: University of Latvia, Institute of Atomic Physics and Spectroscopy, 19 Rainis Boulevard, Riga LV-1586, Latvia.
authors.forename: Alexey
authors.initials: A
authors.lastname: Lihachev
authors.affiliation: Latvian Biomedical Research and Study Centre, Ratsupites iela 1, Riga LV-1067, Latvia.
authors.forename: Ramona
authors.initials: R
authors.lastname: Petrovska
authors.affiliation: University of Latvia, Institute of Atomic Physics and Spectroscopy, 19 Rainis Boulevard, Riga LV-1586, Latvia.
authors.forename: Dainis
authors.initials: D
authors.lastname: Jakovels
authors.affiliation: Vytautas Magnus University, Biophysical Research Group, Faculty of Natural Sciences, Vileikos 8, Kaunas LT-44404, Lithuania.
authors.forename: Saulius
authors.initials: S
authors.lastname: Šatkauskas
mesh_terms: D000818:Animals; D018274:Electroporation / Q000379:methods*; D004867:Equipment Design; D005260:Female; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D006651:Histocytochemistry; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018482:Muscle, Skeletal / Q000378:metabolism*; D061848:Optical Imaging / Q000379:methods*; D013010:Sonication / Q000379:methods*; D014162:Transfection / Q000379:methods
chemical_list: C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins