Selective gene expression analysis of muscular and vascular components in hearts using laser microdissection method.
Selective gene expression analysis of muscular and vascular components in hearts using laser microdissection method.
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DOI：
10.1155/2012/863410
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发表时间：
2012
影响因子：
1.3
通讯作者：
Imaizumi T
中科院分区：
文献类型：
--
作者：
Ikeda A;Kai H;Kajimoto H;Yasuoka S;Kage M;Imaizumi T
背景。心脏由各种细胞组成。然而，单独分析单个成分的基因表达是不可行的。激光显微解剖（LMD）方法是一种从显微区域收集靶细胞的新技术，已被用于恶性肿瘤的治疗。我们试图建立一种从心脏切片中选择性收集肌肉和血管区域的方法，并与该方法比较标记基因的表达。方法与结果。取Wistar-Kyoto大鼠（WKY）和卒中易发自发性高血压大鼠（shrp - sp） 24周龄时的冷冻左心室切片。使用LMD方法，在显微镜引导下选择性地收集肌肉和血管区域。Real-time RT-PCR分析显示，心肌细胞标志物脑型利钠肽（brain-type natriuretic peptide， BNP）在肌肉样品中有表达，而在血管样品中无表达，而平滑肌细胞标志物α-平滑肌肌动蛋白仅在血管样品中有表达。在肌肉组织中，SHR-SP对BNP的上调显著高于WKY （P < 0.05）。结论。LMD方法使我们能够从心肌切片中分别收集肌肉和血管样本，并选择性地评估单个组织成分的mRNA表达。
Background. The heart consists of various kinds of cell components. However, it has not been feasible to separately analyze the gene expression of individual components. The laser microdissection (LMD) method, a new technology to collect target cells from the microscopic regions, has been used for malignancies. We sought to establish a method to selectively collect the muscular and vascular regions from the heart sections and to compare the marker gene expressions with this method. Methods and Results. Frozen left ventricle sections were obtained from Wistar-Kyoto rats (WKY) and stroke-prone spontaneously hypertensive rats (SHR-SP) at 24 weeks of age. Using the LMD method, the muscular and vascular regions were selectively collected under microscopic guidance. Real-time RT-PCR analysis showed that brain-type natriuretic peptide (BNP), a marker of cardiac myocytes, was expressed in the muscular samples, but not in the vascular samples, whereas α-smooth muscle actin, a marker of smooth muscle cells, was detected only in the vascular samples. Moreover, SHR-SP had significantly greater BNP upregulation than WKY (P < 0.05) in the muscular samples. Conclusions. The LMD method enabled us to separately collect the muscular and vascular samples from myocardial sections and to selectively evaluate mRNA expressions of the individual tissue component.