Biotech Research & Innovation Center, University of Copenhagen, Copenhagen, Denmark.
Agilent Technologies Denmark ApS, Glostrup, Denmark.
J Histochem Cytochem. 2023 Sep;71(9):495-508. doi: 10.1369/00221554231194119. Epub 2023 Aug 18.
Recently there have been reports that identify two transient receptor potential channels in cell-matrix junctions known as focal adhesions. These are the calcium channel TRP canonical 7 and the calcium-activated monovalent ion channel, TRP melastatin (TRPM) 4. Here, we report on the occurrence of TRPM4 in focal adhesions of fibroblasts. Of three commercial antibodies recognizing this channel, only one yielded focal adhesion staining, while the other two did not. The epitope recognized by the focal adhesion-localizing antibody was mapped to the extreme C-terminus of the TRPM4 protein. The other two antibodies bind to N-terminal regions of the TRPM4 proteins. Deletion of the gene by CRISPR/cas9 techniques confirmed that this channel is a focal adhesion component, while expression of full-length TRPM4 proteins suggested that processing may occur to yield a form that localizes to focal adhesions. Given the reports that this channel may influence migratory behavior of cells and is linked to cardiovascular disease, TRPM4 functions in adhesion should be explored in greater depth. (J Histochem Cytochem 71: 495-508, 2023).
最近有报道称，在细胞-基质连接处发现了两种瞬时受体电位通道，即称为黏着斑的焦点黏附。这些是钙通道 TRP 经典 7 和钙激活单价离子通道 TRP melastatin（TRPM）4。在这里，我们报告了 TRPM4 在成纤维细胞黏着斑中的存在。在识别该通道的三种商业抗体中，只有一种产生了黏着斑染色，而另外两种则没有。被黏着斑定位抗体识别的表位被映射到 TRPM4 蛋白的极端 C 末端。其他两种抗体结合 TRPM4 蛋白的 N 末端区域。通过 CRISPR/cas9 技术删除 基因证实该通道是 黏着斑的组成部分，而全长 TRPM4 蛋白的表达表明可能会发生加工以产生一种定位于黏着斑的形式。鉴于该通道可能影响细胞的迁移行为并与心血管疾病有关，TRPM4 在黏附中的功能应更深入地研究。（J Histochem Cytochem 71: 495-508, 2023）。
