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In vivo transfer of a neuronal nitric oxide synthase expression vector into the rat bladder by electroporation

Abstract

the isolated bladder and immunohistochemical staining used to detect nNOS; NO x released from isolated bladder strips was also assessed using microdialysis and high-performance liquid chromatography. RESULTSFrom the luciferase assay, 45 V, 1 Hz, 50 ms and eight pulses were selected as the optimum conditions for electroporation. Bladder specimens with GFP genes injected by electroporation showed bright and numerous sites of GFP expression in the smooth muscle layer. In rats with the nNOS gene injected by electroporation there was marked nNOS immunoreactivity, and NO x released from bladder strips was significantly greater than in the control groups. CONCLUSIONSThese results suggest that electroporation is a useful technique for in vivo gene transfer into rat bladder smooth muscles, and that the nNOS gene transferred by this procedure functionally expresses and contributes to NO production.

Nonviral gene transfer to skeletal, smooth, and cardiac muscle in living animals

“…Electroporation-mediated transfer of genes to bladder smooth muscle in rats has also been reported (53). Genes for luciferase, green fluorescent protein (GFP), and neuronal nitric oxide synthase (nNOS) were transferred to the bladder by injecting 50 l of the DNA into the subserosal region of the exposed bladder wall after removal of urine from the bladder.…”

Duration and level of transgene expression after gene electrotransfer to skin in mice

“…[1][2][3] A vast number of studies have encompassed gene electrotransfer to muscles, [4][5][6][7][8] but the technique is also efficient in, for example, cornea, 9 lungs, 10 liver 11,12 kidney, 13 bladder, 14 testis, 15 skin 16,17 and tumor. [18][19][20] One very interesting perspective with gene electrotransfer is that the choice of tissue can determine the duration of expression of the transgene.…”

Herpes simplex virus vector-mediated gene delivery for the treatment of lower urinary tract pain

“…To bypass this barrier, various treatments have been employed; however, their damage to the urothelium would be detrimental to the already damaged urothelium present in IC/PBS patients, and thus would not be of clinical use to improve transduction. Other groups have exposed the bladder by a lower midline incision followed by the direct injection of plasmid DNA expressing a luciferase, eGFP or lacZ reporter gene combined with electroporation to achieve gene delivery, where the voltage, number, duration and frequency of the electric pulses were optimized to provide efficient delivery primarily to cells of the smooth muscle layer 124–126. Moreover, the groups did not detect any difference in KCl-induced detrusor muscle activity following electroporation, suggesting that this approach did not damage the urothelium.…”

“…Otani et al 125. showed that the transduction of bladder by this method with a plasmid encoding the muscarinic M3 receptor did result in detrusor muscle changes that may potentially be therapeutic, whereas Iwashita et al 124. showed that nNOS gene transfer led to an increase in NO production within the bladder smooth muscle layer.…”