Research for the presence and activation mechanism of nitric oxide

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1. The presence of nitric oxide synthase (NOS) in the smooth muscle of genitourinary organs1) Pharmacological analysis :1. Prostate ; Human and canine prostates revealed inhibitory activity via non-adrenergic, non-cholinergic (NANC) nerves using isometric contraction and electrical stimulation techniques.2. Urethra ; Female canine urethra revealed inhibitory activity via NANC nerves using isometric contraction and electrical stimulation techniques. The magnitude of inhibitory activity of the urethra was significantly greater in the proximal region than the distal region, suggesting that NOS nerve may have contribution for urine passage in the proximal urethra, but not in the distal urethra.3. Ureter and urinary bladder ; Human ureter and urinary bladder detrusor revealed inhibitory activity via NANC nerves using isometric contraction and electrical stimulation techniques.2) Biochemical analysis : HPLC with ^3H-arginine and ^3H-citrulline showed that human prostate actually produced NO.3) Immunohistochemical analysis : By using immunohistchemical methods with neuronal NOS (nNOS) antibody, human and canine prostates, human female and female dog urethra, and human ureter revelaed nNOS-positive neuronal structures.4) Molecular biological analysis : Messenger RNA (mRNA) was extracted from human female urethral tissue, and was reversely transcripted using AMV reverse transcriptase. Neuronal NOS DNA was amplified from cDNA of human female urethral tissue by polymerase chain reaction, and two different sizes of the amplified DNA were obtained.2. Localization of NOS in female urethra : Immunohistochemical analysis of localization of nNOS-positive structure revealed the predominance in the proximal region of female dog, but the predominance in the distal region. Conclusion : Prostate and lower urinary tract smoothe muscle, especially urethra, of human and dog possess innervation of nerves with nNOS activity, and inhibitory NANC activity possibly via these nNOS nerves.