Mauser Henry D, Caroza Janessa, Homez Shane Nicole, Arnett Alyssa S, Cutts William D, Lam Daryl W, Thornton Justin, Adams Walter, Kim Brandon J
Department of Biological Sciences, University of Alabama, Tuscaloosa, Alabama, USA.
Department of Biological Sciences, San Jose State University, San Jose, California, USA.
Cell Biochem Funct. 2025 Jun;43(6):e70093. doi: 10.1002/cbf.70093.
Tight junction complexes are crucial features of brain endothelial cells, as they restrict the paracellular route across the blood-brain barrier. Tight junction disruption has been observed in conjunction with numerous diseases of the CNS. In such cases, the organization or integrity of cell-cell junctions may be analyzed with a variety of automated computer programs that quantitatively assess junction images. Here, we directly compare two previously developed python-based programs-JAnaP and IJOQ- for the semi- or fully automated analysis of tight junctions in human stem cell-derived brain-like endothelial cells. Cells were infected with S. pneumoniae and S. agalactiae to initiate junction disruption, and occludin and ZO-1 were analyzed in mock and infected groups via JAnaP and IJOQ. JAnaP and IJOQ both yielded comparable results for the quantification of tight junction disruption in brain endothelial cells. While JAnaP rendered data at the cellular level and gave more information regarding junction phenotype, IJOQ significantly reduced user time and eliminated potential user bias. Our results suggest that JAnaP and IJOQ are both appropriate for quantifying tight junction integrity in brain endothelial cells, and both may offer distinct advantages depending on their context of use.
紧密连接复合体是脑内皮细胞的关键特征，因为它们限制了跨血脑屏障的细胞旁途径。紧密连接破坏已在多种中枢神经系统疾病中被观察到。在这种情况下，可以使用各种自动计算机程序来分析细胞间连接的组织或完整性，这些程序可以定量评估连接图像。在这里，我们直接比较两个先前开发的基于Python的程序——JAnaP和IJOQ——用于对人干细胞衍生的脑样内皮细胞中的紧密连接进行半自动或全自动分析。用肺炎链球菌和无乳链球菌感染细胞以引发连接破坏，并通过JAnaP和IJOQ在未感染组和感染组中分析闭合蛋白和紧密连接蛋白1。JAnaP和IJOQ在定量脑内皮细胞紧密连接破坏方面都产生了可比的结果。虽然JAnaP在细胞水平上提供数据并给出了更多关于连接表型的信息，但IJOQ显著减少了用户时间并消除了潜在的用户偏差。我们的结果表明，JAnaP和IJOQ都适用于定量脑内皮细胞中的紧密连接完整性，并且根据其使用背景两者可能都具有独特的优势。
