Quantification and localization of mRNA levels of endothelin receptor subtypes and preproEndothelin-1 in human urethra

人尿道内皮素受体亚型和前内皮素原-1 mRNA 水平的定量和定位

• 批准号: 12671520
• 负责人: MORIYAMA Nobuo
• 金额: $ 2.18万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12671520/
• 关键词: human urethra; endothelin (ET) receptor; ET receptor mRNA; competitive RT-PCR; in situ hybridization; エンドセリン受容体; エンドセリン受容体mRNA

The vasoconstrictive peptide endothelin-1 (ET-1) is a important peptide in contraction of urethra and uninary bladder. The aim of the present study was to analyze the gene expression of these peptide subtype in human urethra. In this present study I used the competitive reverse transcriptase polymerase chain reaction (RT-PCR) technology and quantified endothelin A (ET_A) and B (ET_B) receptor subtype mRNAs and preproET-1 mRNA levels using the technique reported by Asberg et al.(Scand J Clin Lab Invest 58:299-306, 1998). Furthermore I used the in situ hybridization method to show the localization of these mRNA in human urethra. The probes for competitive RT-PCR and in situ hybridization (labeled with Alexa fluor 488) are mentioned below.【table】The whole urethral specimens obtained in total cysturethrectomy for the female and male patients with bladder tumor were divided for 3-4 pieces. In all specimens, mRNA levels of ET_A, ET_B receptor subtype mRNAs and preproET-1 mRNA levels were 3 x 10^5〜10^6 competitor contents (copy) and almost same amounts. The localization of above probes was in the smooth muscle cells. There is no difference in labeled areas for the probes.

血管收缩肽内皮素-1 （ET-1）是尿道和膀胱收缩的重要肽。本研究的目的是分析这些肽亚型在人类尿道中的基因表达。在本研究中，我使用竞争性逆转录酶聚合酶链式反应（RT-PCR）技术，并使用Asberg等人报道的技术定量内皮素A （ET_A）和B （ET_B）受体亚型mRNA和preproET-1 mRNA水平（Scand J clinical Lab Invest 58:299-306, 1998）。此外，我使用原位杂交方法显示这些mRNA在人类尿道中的定位。竞争性RT-PCR探针和原位杂交探针（用Alexa flu488标记）如下所述。【表】男女膀胱肿瘤患者全膀胱切除术获得的全尿道标本分为3-4片。在所有标本中，ET_A、ET_B受体亚型mRNA和preproET-1 mRNA水平均为3 × 10^5 ~ 10^6竞争对手含量（拷贝），且数量基本相同。上述探针均定位于平滑肌细胞。探针的标记区域没有区别。

项目成果

Usui, T., Hara, M., Satoh, H., Moriyama, N., et al.: "Molecular basis of ocular abnormalities associated with proximal renal tubular acidosis"J. Clin. Invest.. 108. 107-115 (2001)

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Shindo，T.，Kuriharal，H.，Kuno，K.，横山，H.，和田，T.，栗原，Y.，今井，T.，王，Y.，绪方，M.，西松，H.，

Shindo, T., Kurihara1, H., Kuno, K., Yokoyama, H., Wada, T., Kurihara, Y., Imai, T., Wang, Y.: "ADAMTS-1 : a metalloproteinase-disintegrin essential for normal growth, fertility, and organ morphology and function"J. Clin. Invest.. 105. 1345-1352 (2000)

Shindo, T.、Kurihara1, H.、Kuno, K.、Yokoyama, H.、Wada, T.、Kurihara, Y.、Imai, T.、Wang, Y.：“ADAMTS-1：金属蛋白酶解整合素必需的

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