Dean A D, Vehaskari V M, Greenwald J E
Department of Medicine, Washington University School of Medicine, St. Louis, Missouri 63110.
Am J Physiol. 1994 Mar;266(3 Pt 2):F491-6. doi: 10.1152/ajprenal.1994.266.3.F491.
We have previously demonstrated the synthesis of atrial natriuretic factor (ANF) in the distal cortical nephron of the rat kidney. We now report the synthesis of C-type natriuretic peptide (CNP) in the rat, mouse, and human nephron. CNP mRNA was initially detected in the rat and mouse kidney, as well as in three transformed cell lines isolated from the proximal and distal nephron, using reverse transcription-polymerase chain reaction (RT-PCR). Confirmation of the kidney PCR product was performed by nucleotide sequence analysis and Southern hybridization. Northern hybridization of rat brain CNP cDNA to human kidney polyadenylated RNA detected a 1.4-kb gene transcript. Rat nephron segments were microdissected and subjected to RT-PCR to localize CNP mRNA. CNP mRNA was detected in the proximal convoluted tubule, cortical collecting duct, medullary thick limbs, and inner medullary collecting ducts. CNP, as detected by immunohistochemistry, was found to colocalize with CNP mRNA.
我们先前已证明大鼠肾脏远曲皮质肾单位可合成心房利钠因子(ANF)。我们现在报告在大鼠、小鼠和人类肾单位中C型利钠肽(CNP)的合成情况。使用逆转录-聚合酶链反应(RT-PCR),最初在大鼠和小鼠肾脏以及从近端和远端肾单位分离出的三种转化细胞系中检测到CNP mRNA。通过核苷酸序列分析和Southern杂交对肾脏PCR产物进行确认。用大鼠脑CNP cDNA与人类肾脏多聚腺苷酸化RNA进行Northern杂交,检测到一个1.4 kb的基因转录本。对大鼠肾单位节段进行显微切割,并进行RT-PCR以定位CNP mRNA。在近端曲管、皮质集合管、髓袢升支粗段和内髓集合管中检测到CNP mRNA。通过免疫组织化学检测发现,CNP与CNP mRNA共定位。