Fischer Anthony J, Goss Kelli L, Scheetz Todd E, Wohlford-Lenane Christine L, Snyder Jeanne M, McCray Paul B
Department of Pediatrics, University of Iowa, Iowa City, IA 52242, USA.
Am J Respir Cell Mol Biol. 2009 Feb;40(2):189-99. doi: 10.1165/rcmb.2008-0240OC. Epub 2008 Aug 14.
Human conducting airways contain two anatomically distinct epithelial cell compartments: surface epithelium and submucosal glands (SMG). Surface epithelial cells interface directly with the environment and function in pathogen detection, fluid and electrolyte transport, and mucus elevation. SMG secrete antimicrobial molecules and most of the airway surface fluid. Despite the unique functional roles of surface epithelia and SMG, little is known about the differences in gene expression and cellular metabolism that orchestrate the specialized functions of these epithelial compartments. To approach this problem, we performed large-scale transcript profiling using epithelial cell samples obtained by laser capture microdissection (LCM) of human bronchus specimens. We found that SMG expressed high levels of many transcripts encoding known or putative innate immune factors, including lactoferrin, zinc alpha-2 glycoprotein, and proline-rich protein 4. By contrast, surface epithelial cells expressed high levels of genes involved in basic nutrient catabolism, xenobiotic clearance, and ciliated structure assembly. Selected confirmation of differentially expressed genes in surface and SMG epithelia demonstrated the predictive power of this approach in identifying genes with localized tissue expression. To characterize metabolic differences between surface epithelial cells and SMG, immunostaining for a mitochondrial marker (isocitrate dehydrogenase) was performed. Because greater staining was observed in the surface compartment, we predict that these cells use significantly more energy than SMG cells. This study illustrates the power of LCM in defining the roles of specific anatomic features in airway biology and may be useful in examining how disease states alter transcriptional programs in the conducting airways.
表面上皮和黏膜下腺(SMG)。表面上皮细胞直接与外界环境接触,在病原体检测、液体和电解质运输以及黏液提升方面发挥作用。黏膜下腺分泌抗菌分子和大部分气道表面液体。尽管表面上皮和黏膜下腺具有独特的功能作用,但对于协调这些上皮区室特殊功能的基因表达和细胞代谢差异却知之甚少。为了解决这个问题,我们使用通过对人支气管标本进行激光捕获显微切割(LCM)获得的上皮细胞样本进行了大规模转录谱分析。我们发现黏膜下腺表达了许多编码已知或推定的固有免疫因子的转录本,包括乳铁蛋白、锌α-2糖蛋白和富含脯氨酸的蛋白4。相比之下,表面上皮细胞表达了参与基本营养物质分解代谢、外源性物质清除和纤毛结构组装的高水平基因。对表面和黏膜下腺上皮中差异表达基因的选定验证证明了这种方法在鉴定具有局部组织表达的基因方面的预测能力。为了表征表面上皮细胞和黏膜下腺之间的代谢差异,对线粒体标记物(异柠檬酸脱氢酶)进行了免疫染色。由于在表面区室观察到更强的染色,我们预测这些细胞比黏膜下腺细胞消耗的能量要多得多。这项研究说明了激光捕获显微切割在定义气道生物学中特定解剖特征的作用方面的能力,并且可能有助于研究疾病状态如何改变传导气道中的转录程序。