Nebigil C G, Garnovskaya M N, Spurney R F, Raymond J R
Department of Medicine, Duke University, Durham, North Carolina.
Am J Physiol. 1995 Jan;268(1 Pt 2):F122-7. doi: 10.1152/ajprenal.1995.268.1.F122.
Previous studies have demonstrated the presence of mitogenic serotonin [i.e., 5-hydroxytryptamine (5-HT)] receptors on glomerular mesangial cells and have linked those receptors to a complicated array of intracellular and autocrine/paracrine signaling pathways [T. Knauss and H. E. Abboud. Am. J. Physiol. 251 (Renal Fluid Electrolyte Physiol. 20): F844-F850, 1986; and N. Takuwa, M. Ganz, Y. Takuwa, R. B. Sterzel, and H. Rasmussen. Am. J. Physiol. 257 (Renal Fluid Electrolyte Physiol. 26): F431-F439, 1989]. Those studies suggested that the mesangial subtype of 5-HT receptor is a member of the 5-HT2 receptor family, which consists of three known members, designated as subtypes A, B, and C. The purpose of the current study was to identify the subtype of 5-HT2 receptor present on mesangial cells. Northern blot showed detectable mRNA for a putative 5-HT2A receptor, but not for 5-HT1A or 5-HT2C receptors. Reverse transcription-polymerase chain reaction (RT-PCR) with degenerate oligonucleotides derived from the putative third and sixth transmembrane domains of cloned 5-HT2 receptors yielded a 580-nucleotide (nt) fragment. RT-PCR with primers highly specific for the 5-HT2A receptor and designed to amplify > 95% of its coding block yielded a product of 1,320 nt. Nested PCR reactions yielded products of the predicted sizes for the 5-HT2A receptor. Partial sequence information was obtained, and the sequence corresponded exactly (627/627 nt) to that published for the cloned rat brain 5-HT2A receptor. These studies identify the mesangial cell mitogenic 5-HT receptor as a 5-HT2A receptor subtype.
以往的研究已证实肾小球系膜细胞上存在促有丝分裂的血清素[即5-羟色胺（5-HT）]受体，并将这些受体与一系列复杂的细胞内及自分泌/旁分泌信号通路联系起来[T. 克瑙斯和H. E. 阿博德。《美国生理学杂志》251卷（肾液体电解质生理学20）：F844 - F850，1986年；以及N. 卓和、M. 甘茨、Y. 卓和、R. B. 施特尔策尔和H. 拉斯穆森。《美国生理学杂志》257卷（肾液体电解质生理学26）：F431 - F439，1989年]。这些研究表明5-HT受体的系膜细胞亚型是5-HT2受体家族的成员，该家族由三个已知成员组成，分别命名为A、B和C亚型。本研究的目的是鉴定系膜细胞上存在的5-HT2受体亚型。Northern印迹显示可检测到推测的5-HT2A受体的mRNA，但未检测到5-HT1A或5-HT2C受体的mRNA。用源自克隆的5-HT2受体推测的第三和第六跨膜结构域的简并寡核苷酸进行逆转录-聚合酶链反应（RT-PCR），产生了一个580个核苷酸（nt）的片段。用对5-HT2A受体高度特异且设计用于扩增其>95%编码区的引物进行RT-PCR，产生了一个1320 nt的产物。巢式PCR反应产生了5-HT2A受体预测大小的产物。获得了部分序列信息，该序列与已发表的克隆大鼠脑5-HT2A受体的序列完全一致（627/627 nt）。这些研究将系膜细胞促有丝分裂的5-HT受体鉴定为5-HT2A受体亚型。
