Expression of Functional Cell-Cell Channels from Cloned Rat Liver Gap Junction Complementary DNA

• DOI: 10.1126/science.3035715
• Publication Date: 2006-10-05T20:28:58Z
• Authors: G. Dahl; T. Miller; D. Paul; R. Voellmy; R. Werner
• Keywords: 0303 health sciences; Transcription, Genetic; Xenopus; Membrane Proteins; Nucleic Acid Hybridization; DNA; Connexins; Rats; 03 medical and health sciences; Intercellular Junctions; Liver; Oocytes; Animals; RNA, Messenger; Cloning, Molecular; Promoter Regions, Genetic; Heat-Shock Proteins

An oocyte expression system was used to test the relation between a complementary DNA (cDNA) clone encoding the liver gap junction protein and cell-cell channels. Total liver polyadenylated messenger RNA injected into oocytes induced cell-cell channels between paired oocytes. This induction was blocked by simultaneous injection of antisense RNA transcribed from the gap junction cDNA. Messenger RNA selected by hybridization to the cDNA clone and translated in oocyte pairs yielded a higher junctional conductance than unselected liver messenger RNA. Cell-cell channels between oocytes were also formed when the cloned cDNA was expressed under the control of a heat-shock promoter. A concentration-dependent induction of channels was observed in response to injection with in vitro transcribed gap junction messenger RNA. Thus, the liver gap junction cDNA encodes a protein that is essential for the formation of functional cell-cell channels.