{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,16]],"date-time":"2026-10-16T22:16:51Z","timestamp":1792189011458,"version":"4.4.0"},"reference-count":69,"publisher":"Wiley","issue":"8","license":[{"start":{"date-parts":[[2017,2,14]],"date-time":"2017-02-14T00:00:00Z","timestamp":1487030400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["NG 4\/10\u20101"],"award-info":[{"award-number":["NG 4\/10\u20101"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["The Journal of Physiology"],"published-print":{"date-parts":[[2017,4,15]]},"abstract":"<jats:sec><jats:title>Key points<\/jats:title><jats:p><jats:list list-type=\"bullet\"><jats:list-item><jats:p>Gap junction channels are essential for the formation and regulation of physiological units in tissues by allowing the lateral cell\u2010to\u2010cell diffusion of ions, metabolites and second messengers.<\/jats:p><\/jats:list-item><jats:list-item><jats:p>Stimulation of the adenosine receptor subtype A<jats:sub>2B<\/jats:sub>increases the gap junction coupling in the human blood\u2013brain barrier endothelial cell line hCMEC\/D3.<\/jats:p><\/jats:list-item><jats:list-item><jats:p>Although the increased gap junction coupling is cAMP\u2010dependent, neither the protein kinase A nor the exchange protein directly activated by cAMP were involved in this increase.<\/jats:p><\/jats:list-item><jats:list-item><jats:p>We found that cAMP activates cyclic nucleotide\u2010gated (CNG) channels and thereby induces a Ca<jats:sup>2+<\/jats:sup>influx, which leads to the increase in gap junction coupling.<\/jats:p><\/jats:list-item><jats:list-item><jats:p>The report identifies CNG channels as a possible physiological link between adenosine receptors and the regulation of gap junction channels in endothelial cells of the blood\u2013brain barrier.<\/jats:p><\/jats:list-item><\/jats:list><\/jats:p><\/jats:sec><jats:sec><jats:title>Abstract<\/jats:title><jats:p>The human cerebral microvascular endothelial cell line hCMEC\/D3 was used to characterize the physiological link between adenosine receptors and the gap junction coupling in endothelial cells of the blood\u2013brain barrier. Expressed adenosine receptor subtypes and connexin (Cx) isoforms were identified by RT\u2010PCR. Scrape loading\/dye transfer was used to evaluate the impact of the A<jats:sub>2A<\/jats:sub>and A<jats:sub>2B<\/jats:sub>adenosine receptor subtype agonist 2\u2010phenylaminoadenosine (2\u2010PAA) on the gap junction coupling. We found that 2\u2010PAA stimulated cAMP synthesis and enhanced gap junction coupling in a concentration\u2010dependent manner. This enhancement was accompanied by an increase in gap junction plaques formed by Cx43. Inhibition of protein kinase A did not affect the 2\u2010PAA\u2010related enhancement of gap junction coupling. In contrast, the cyclic nucleotide\u2010gated (CNG) channel inhibitor<jats:sc>l<\/jats:sc>\u2010<jats:italic>cis<\/jats:italic>\u2010diltiazem, as well as the chelation of intracellular Ca<jats:sup>2+<\/jats:sup>with BAPTA, or the absence of external Ca<jats:sup>2+<\/jats:sup>, suppressed the 2\u2010PAA\u2010related enhancement of gap junction coupling. Moreover, we observed a 2\u2010PAA\u2010dependent activation of CNG channels by a combination of electrophysiology and pharmacology. In conclusion, the stimulation of adenosine receptors in hCMEC\/D3 cells induces a Ca<jats:sup>2+<\/jats:sup>influx by opening CNG channels in a cAMP\u2010dependent manner. Ca<jats:sup>2+<\/jats:sup>in turn induces the formation of new gap junction plaques and a consecutive sustained enhancement of gap junction coupling. The report identifies CNG channels as a physiological link that integrates gap junction coupling into the adenosine receptor\u2010dependent signalling of endothelial cells of the blood\u2013brain barrier.<\/jats:p><\/jats:sec>","DOI":"10.1113\/jp273150","type":"journal-article","created":{"date-parts":[[2017,1,11]],"date-time":"2017-01-11T12:28:12Z","timestamp":1484137692000},"page":"2497-2517","source":"Crossref","is-referenced-by-count":22,"title":["Adenosine receptors regulate gap junction coupling of the human cerebral microvascular endothelial cells hCMEC\/D3 by Ca<sup>2+<\/sup>influx through cyclic nucleotide\u2010gated channels"],"prefix":"10.1113","volume":"595","author":[{"given":"Almke","family":"Bader","sequence":"first","affiliation":[{"name":"Institute of Biophysics Leibniz University Hannover Hannover Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Willem","family":"Bintig","sequence":"additional","affiliation":[{"name":"Institute of Biochemistry 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