Abstract：
Connexins are essential gap junction proteins that play pivotal roles in intercellular communication in various organs of mammals. Connexin-43 (Cx43) is expressed in various components of the immune system, and there is extensive evidence of its participation in inflammation responses. The involvement of Cx43 in macrophage functionality involves the purinergic signaling pathway. Macrophages contribute to defenses against inflammatory reactions such as bacterial sepsis and peritonitis. Several assays can identify the presence and activity of Cx43 in macrophages. Real-time polymerase chain reaction (PCR) can measure the relative mRNA expression of Cx43, whereas western blotting can detect protein expression levels. Using immunofluorescence assays, it is possible to analyze the expression and observe the localization of Cx43 in cells or tissues. Moreover, connexin-mediated gap junction intercellular communication can be evaluated using functional assays such as microinjection of fluorescent dyes or scrape loading-dye transfer. The use of selective inhibitors contributes to this understanding and reinforces the role of connexins in various processes. Here, we discuss these methods to evaluate Cx43 and macrophage gap junctions.
摘要：
连接蛋白是必需的间隙连接蛋白，在哺乳动物各种器官的细胞间通讯中起关键作用。连接蛋白-43（Cx43）在免疫系统的各种成分中表达，并且有大量证据表明其参与炎症反应。Cx43参与巨噬细胞功能涉及嘌呤能信号通路。巨噬细胞有助于防御炎症反应,如细菌性败血症和腹膜炎。几种测定可以鉴定巨噬细胞中Cx43的存在和活性。实时聚合酶链反应（PCR）可以测量Cx43的相对mRNA表达，而Western印迹可以检测蛋白质表达水平。使用免疫荧光分析，分析Cx43在细胞或组织中的表达并观察其定位是可能的。此外，连接蛋白介导的间隙连接细胞间通讯可以使用功能测定来评估，例如荧光染料的显微注射或刮擦负载-染料转移。选择性抑制剂的使用有助于这种理解并加强连接蛋白在各种过程中的作用。这里,我们讨论了这些方法来评估Cx43和巨噬细胞间隙连接。
