Optimizing in vivo gene transfer into mouse corpus cavernosum by use of surface electroporation.
10.4111/kju.2015.56.3.197
- Author:
Kang Moon SONG
1
;
Min Ji CHOI
;
Mi Hye KWON
;
Kalyan GHATAK
;
Soo Hwan PARK
;
Dong Soo RYU
;
Ji Kan RYU
;
Jun Kyu SUH
Author Information
1. National Research Center for Sexual Medicine and Department of Urology, Inha University School of Medicine, Incheon, Korea. jksuh@inha.ac.kr
- Publication Type:Original Article ; Research Support, Non-U.S. Gov't
- Keywords:
Electroporation;
Erectile dysfunction;
Gene therapy;
Penis
- MeSH:
Animals;
Diabetes Mellitus, Experimental/complications;
Electroporation/*methods;
Erectile Dysfunction/*therapy;
Gene Expression;
Gene Transfer Techniques;
Genes, Reporter;
Genetic Therapy/*methods;
Luciferases/metabolism;
Male;
Mice;
Mice, Inbred C57BL;
Penile Erection/physiology;
Penis/*physiopathology;
Transfection
- From:Korean Journal of Urology
2015;56(3):197-204
- CountryRepublic of Korea
- Language:English
-
Abstract:
PURPOSE: Electroporation is known to enhance the efficiency of gene transfer through a transient increase in cell membrane permeability. The aim of this study was to determine the optimal conditions for in vivo electroporation-mediated gene delivery into mouse corpus cavernosum. MATERIALS AND METHODS: Diabetes was induced in C57BL/6 mice by intraperitoneal injections of streptozotocin. After intracavernous injection of pCMV-Luc (100 microg/40 microL), different electroporation settings (5-50 V, 8-16 pulses with a duration of 40-100 ms) were applied to the penis to establish the optimal conditions for electroporation. Gene expression was evaluated by luciferase assay. We also assessed the undesired consequences of electroporation by visual inspection and hematoxylin-eosin staining of penile tissue. RESULTS: Electroporation profoundly induced gene expression in the corpus cavernosum tissue of normal mice in a voltage-dependent manner. We observed electrical burn scars in the penis of normal mice who received electroporation with eight 40-ms pulses at a voltage of 50 V and sixteen 40-ms pulses, eight 100-ms pulses, and sixteen 100-ms pulses at a voltage of 30 V. No detectable burn scars were noted in normal mice stimulated with eight 40-ms pulses at a voltage of 30 V. Electroporation also significantly induced gene expression in diabetic mice stimulated with 40-ms pulse at a voltage of 30 V without injury to the penis. CONCLUSIONS: We have established the optimal electroporation conditions for maximizing gene transfer into the corpus cavernosum of mice while avoiding damage to the erectile tissue. The electroporation-mediated gene delivery technique will be a valuable tool for gene therapy in the field of erectile dysfunction.
