Vanderbilt O'Brien Center, Department of Medicine, Vanderbilt University School of Medicine, Nashville, Tennessee, USA.
J Histochem Cytochem. 2010 May;58(5):413-20. doi: 10.1369/jhc.2009.954966. Epub 2010 Jan 19.
Tissue microarray (TMA) is a new high-throughput method that enables simultaneous analysis of the profiles of protein expression in multiple tissue samples. TMA technology has not previously been adapted for physiological and pathophysiological studies of rodent kidneys. We have evaluated the validity and reliability of using TMA to assess protein expression in mouse and rat kidneys. A representative TMA block that we have produced included: (1) mouse and rat kidney cortex, outer medulla, and inner medulla fixed with different fixatives; (2) rat kidneys at different stages of development fixed with different fixatives; (3) mouse and rat kidneys with different physiological or pathophysiological treatments; and (4) built-in controls. As examples of the utility, immunostaining for cyclooxygenase-2, renin, Tamm Horsfall protein, aquaporin-2, connective tissue growth factor, and synaptopodin was carried out with kidney TMA slides. Quantitative analysis of cyclooxygense-2 expression in kidneys confirms that individual cores provide meaningful representations comparable to whole-kidney sections. These studies show that kidney TMA technique is a promising and useful tool for investigating the expression profiles of proteins of interest in rodent kidneys under different physiological and pathophysiological conditions.
组织微阵列(TMA)是一种高通量方法,可实现对多个组织样本中蛋白质表达谱的同时分析。TMA 技术以前尚未应用于啮齿动物肾脏的生理和病理生理学研究。我们评估了使用 TMA 评估小鼠和大鼠肾脏中蛋白质表达的有效性和可靠性。我们制作的代表性 TMA 组织块包括:(1)用不同固定剂固定的小鼠和大鼠肾脏皮质、外髓质和内髓质;(2)用不同固定剂固定的不同发育阶段的大鼠肾脏;(3)用不同生理或病理生理处理的小鼠和大鼠肾脏;和(4)内置对照。作为实用性的示例,用肾脏 TMA 载玻片进行了环氧化酶-2、肾素、Tamm Horsfall 蛋白、水通道蛋白-2、结缔组织生长因子和突触蛋白聚糖的免疫染色。对肾脏中环氧化酶-2 表达的定量分析证实,单个核心提供了有意义的代表性,与整个肾脏切片相当。这些研究表明,肾脏 TMA 技术是一种有前途且有用的工具,可用于研究不同生理和病理生理条件下啮齿动物肾脏中感兴趣的蛋白质的表达谱。