Antibodies targeting extracellular domain of connexins for studies of hemichannels

Hemichannels are transmembrane channels composed of either a connexin or pannexin hexamer. The docking of the extracellular domains of connexin hemichannels contributed by neighboring cells forms a gap junction channel that joins the cytoplasm of adjacent cells. Connexins are expressed ubiquitously in different organs, but some subtypes are expressed exclusively in certain tissues and tumors. Both gap junction channels and hemichannels participate in diverse physiological and pathological responses. However, the lack of specific reagents that inhibit only gap junction channels or hemichannels is a challenge that makes it different to discern the specific roles of either channel. Fortunately, the available information regarding the connexin sequence, secondary and tertiary structure, and their biochemical and physiological properties permits the development of strategies to block exclusively the hemichannel activity exclusively, with no effect on gap junction activity. This task is accomplished through the use of specifics antibodies that target the extracellular sites of desired connexin subtype. However, the underlying mechanism of how antibodies targeting extracellular connexin epitopes actually inhibit hemichannels remains unknown. Although these antibodies are being used for detecting and blocking of hemichannels in normal and tumor cells, they can also be potentially used for tissue-specific treatment and drug delivery in clinical applications. In this article, we will first review the literature concerning the structure of connexins and the unique properties of extracellular loop domains of the connexins. Furthermore, we will discuss briefly the development of connexin (Cx) 43(E2) antibody, a specific antibody which detects the second extracellular loop of Cx43 and specifically prevents the opening of Cx43 hemichannels. We will then summarize the reported studies of specific reagents used for the inhibition of connexin hemichannels including antibodies developed against extracellular loop domains.This article is part of the Special Issue Section entitled ‘Current Pharmacology of Gap Junction Channels and Hemichannels’.

► Overview of structures of connexins, gap junctions and hemichannels.
► A major challenge in development of reagent that block hemichannels, but not gap junctions.
► Review antibodies against Cx43 extracellular loop domains and applications.
► Process of generating a specific antibody against 2nd extracellular loop of Cx43 and application.
► Research advances and future perspectives of development and application of blocking antibody.