Yamaguchi S, Tamura K, Nyui N, Hibi K, Ishigami T, Kihara M, Yabana M, Sesoko S, Ishii M, Umemura S
Second Department of Internal Medicine, Yokohama City University School of Medicine, Japan.
Hypertens Res. 1998 Sep;21(3):155-61. doi: 10.1291/hypres.21.155.
We studied the localization of angiotensinogen mRNA in rat nephron segments and the differences in angiotensinogen mRNA levels between male Sprague-Dawley rats at 6 and 12 wk of age using reverse transcription and polymerase chain reaction (RT-PCR). Each nephron segment of the rat kidney was microdissected. Total RNA was prepared and used in the following RT-PCR assay. The PCR products were size-fractionated by agarose gel electrophoresis, visualized with ethidium bromide staining, and identified by Southern blot analysis. The relative amounts of products were determined by densitometry. Strong bands corresponding to angiotensinogen mRNA were detected from proximal convoluted and straight tubules, and weaker bands were found in glomeruli. The signals in all tissues in 12-wk-old rats were weaker than those in 6-wk-old rats. Since local angiotensinogen is the unique substrate of the tissue renin-angiotensin system and exerts an autocrine-paracrine influence on renal function, the changes in tubular angiotensinogen may be related to physiological and morphological changes in the rat kidney during development.
我们使用逆转录聚合酶链反应（RT-PCR）研究了血管紧张素原mRNA在大鼠肾单位各节段中的定位，以及6周龄和12周龄雄性斯普拉格-道利大鼠之间血管紧张素原mRNA水平的差异。对大鼠肾脏的每个肾单位节段进行显微切割。制备总RNA并用于以下RT-PCR检测。PCR产物通过琼脂糖凝胶电泳进行大小分级，用溴化乙锭染色可视化，并通过Southern印迹分析进行鉴定。产物的相对量通过光密度测定法确定。在近端曲管和直小管中检测到与血管紧张素原mRNA相对应的强条带，在肾小球中发现较弱的条带。12周龄大鼠所有组织中的信号均弱于6周龄大鼠。由于局部血管紧张素原是组织肾素-血管紧张素系统的唯一底物，并对肾功能发挥自分泌-旁分泌作用，肾小管血管紧张素原的变化可能与大鼠肾脏发育过程中的生理和形态变化有关。
