Research for the presence and activation mechanism of nitric oxide

1. 一氧化氮合酶（NOS）在泌尿生殖器官平滑肌中的存在前列腺癌;通过等长收缩和电刺激技术，人类和犬前列腺通过非肾上腺素能、非胆碱能（NANC）神经显示抑制活性。尿道;通过等长收缩和电刺激技术，雌性犬尿道通过NANC神经显示抑制活性。尿道近端抑制活性的幅度明显大于远端，提示NOS神经可能对尿道近端尿路有贡献，而对远端尿路无贡献。输尿管和膀胱；利用等长收缩和电刺激技术，人类输尿管和膀胱逼尿肌通过NANC神经显示出抑制活性。2)生化分析：用^ 3h -精氨酸和^ 3h -瓜氨酸高效液相色谱法显示人前列腺确实产生NO.3)免疫组化分析：用神经元NOS （nNOS）抗体免疫组化方法，人、犬前列腺、人雌、犬尿道、人输尿管均发现nNOS阳性的神经元结构。4)分子生物学分析：从人类女性尿道组织中提取信使RNA (mRNA)，利用AMV逆转录酶进行逆转录。用聚合酶链反应从人女性尿道组织cDNA中扩增神经元NOS DNA，得到两种不同大小的扩增DNA。雌性尿道NOS的定位：免疫组化分析显示，雌性犬的NOS阳性结构定位以近端区为主，远端区为主。结论：人和狗的前列腺和下尿路平滑肌，特别是尿道中存在nNOS活性神经的支配，其抑制NANC活性可能是通过这些nNOS神经。

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.