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1)药理分析：1.前列腺：人和狗的前列腺通过非肾上腺素能、非胆碱能(NANC)神经等长收缩和电刺激技术显示出抑制活性。利用等长收缩和电刺激技术，雌性犬的尿路通过NANC神经表现出抑制活动。近端对尿路的抑制作用明显大于远端，提示一氧化氮合酶神经可能在近端对尿路有贡献，而在远端对尿路无影响。2)生化分析：经3 H-精氨酸和3 H-瓜氨酸的高效液相色谱检测表明，人前列腺实际上产生了3)免疫组织化学分析：3)免疫组织化学分析：用神经元型一氧化氮合酶(NNOS)抗体，人和狗的前列腺、人的雌性和雌性犬的尿道以及人的输尿管显示出nNOS阳性的神经元结构。4)分子生物学分析：从人女性的尿路组织中提取信使RNA(信使RNA)，并用AMV逆转录酶进行逆转录。方法：1.采用聚合酶链式反应从人女性尿路组织中扩增出神经元型一氧化氮合酶DNA，扩增出两种不同大小的DNA。雌性犬尿路中一氧化氮合酶的定位：免疫组织化学分析显示，雌性犬尿路中神经元型一氧化氮合酶阳性结构以近端为主，远端以远端为主。结论：人和狗的前列腺和下尿路的肌肉，尤其是尿路具有nNOS神经的神经支配，nNOS神经可能通过这些神经抑制了NANC的活动。

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.