Yoshida Takumi, Müller Eva, Stears Robin, Shirota Satsuki, Tsuchiya Ken, Akiba Takashi, Gullans Steven R
Department of Blood Purification, Kidney Center, Tokyo Women's Medical University, Tokyo, Japan.
Biochem Biophys Res Commun. 2004 Sep 10;322(1):250-7. doi: 10.1016/j.bbrc.2004.07.104.
To distinguish biological molecular processes of osmotic stress occurring in inner medulla, we utilized microarrays to monitor expression profiles. RNAs from three segments (cortex, outer medulla, and inner medulla) of mouse kidney were isolated and applied to microarrays. We found 35 genes expressed highly in inner medulla. Next, microarrays for the RNAs from mouse medullary collecting duct cell line (mIMCD) cells and osmotically adapted mIMCD cells (HT cells) were performed (designed as resistant to 1270mOsm/H(2)O). Of 35 genes highly expressed in inner medulla, 6 genes such as; B-cell translocation gene protein (BTG), myc-basic motif homologue, gelsolin, cell surface glycoprotein, laminin beta2, and tubulo-interstitial nephritis antigen, were also expressed highly in HT cells. Using real-time PCR, we confirmed the expression of six genes. Additionally acute osmotic stress induced the BTG. By comparing the inner medulla to a mIMCD3, we identified genes which respond to acute and chronic hyperosmotic stress.
为了区分发生在内髓质的渗透应激的生物分子过程,我们利用微阵列来监测表达谱。从小鼠肾脏的三个部分(皮质、外髓质和内髓质)分离RNA并应用于微阵列。我们发现35个基因在内髓质中高表达。接下来,对来自小鼠髓质集合管细胞系(mIMCD)细胞和渗透适应的mIMCD细胞(HT细胞)的RNA进行微阵列分析(设计为对1270mOsm/H₂O有抗性)。在内髓质中高表达的35个基因中,6个基因,如B细胞易位基因蛋白(BTG)、myc碱性基序同源物、凝溶胶蛋白、细胞表面糖蛋白、层粘连蛋白β2和肾小管间质性肾炎抗原,在HT细胞中也高表达。我们使用实时PCR证实了这6个基因的表达。此外,急性渗透应激诱导了BTG。通过将内髓质与mIMCD3进行比较,我们鉴定出了对急性和慢性高渗应激有反应的基因。