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

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尿道中的基因表达。本研究采用竞争性逆转录聚合酶链反应（RT-PCR）技术，采用Asberg等报道的方法定量测定内皮素A（ET_A）和B（ET_B）受体亚型mRNA和前体ET-1 mRNA水平。（Scand J Clin Lab Invest 58：299-306，1998）。并利用原位杂交方法显示了这些mRNA在人尿道中的定位。用于竞争性RT-PCR和原位杂交的探针（用Alexa fluor 488标记）如下所述。[表]对男女膀胱肿瘤患者行全膀胱尿道切除术，取全尿道标本，分为3-4块。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竞争物含量（拷贝），且几乎相同。上述探针均定位于平滑肌细胞内。探针的标记面积无差异。

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.