LRRCA8A and ANO1 contribute to serum-induced VRAC in a Ca2+-dependent manners
The volume-regulated anion channel (VRAC) plays a central role in maintaining cell volume in response to osmotic stress. Leucine-rich repeat-containing 8A (LRRC8A) was recently identified as an essential component of VRAC although other Cl− channels were also suggested to contribute to VRAC. VRAC is...
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Elsevier
2020-07-01
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| Series: | Journal of Pharmacological Sciences |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S1347861320300384 |
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| author | Huiran Zhang Yani Liu Hongchao Men Fan Zhang Hailin Zhang |
| author_facet | Huiran Zhang Yani Liu Hongchao Men Fan Zhang Hailin Zhang |
| author_sort | Huiran Zhang |
| collection | DOAJ |
| description | The volume-regulated anion channel (VRAC) plays a central role in maintaining cell volume in response to osmotic stress. Leucine-rich repeat-containing 8A (LRRC8A) was recently identified as an essential component of VRAC although other Cl− channels were also suggested to contribute to VRAC. VRAC is activated when a cell is challenged with a hypotonic environment or even in isotonic conditions challenged with different stimuli. It is not clear how VRAC is activated and whether activation of VRAC in hypotonic and isotonic conditions share the same mechanism. In this present study, we investigated relative contribution of LRRC8A and anoctamin 1(ANO1) to VRAC currents activated by fetal bovine serum (FBS) in isotonic condition, and studied the role of intracellular Ca2+ in this activation. We used CRISPR/Cas9 gene editing approach, electrophysiology, and pharmacology approaches to show that VRAC currents induced by FBS is mostly mediated by LRRC8A in HEK293 cells, but also with significant contribution from ANO1. FBS induces Ca2+ transients and these Ca2+ signals are required for the activation of VRAC by serum. These findings will help to further understand the mechanism in activation of VRAC. |
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| institution | Directory Open Access Journal |
| issn | 1347-8613 |
| language | English |
| last_indexed | 2024-12-22T13:05:14Z |
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| spelling | doaj.art-cde4fc99bc274060b139d6cdf87a47a12022-12-21T18:24:54ZengElsevierJournal of Pharmacological Sciences1347-86132020-07-011433176181LRRCA8A and ANO1 contribute to serum-induced VRAC in a Ca2+-dependent mannersHuiran Zhang0Yani Liu1Hongchao Men2Fan Zhang3Hailin Zhang4Department of Pharmacology, Hebei Medical University, The Key Laboratory of Neural and Vascular Biology, Ministry of Education, Shijiazhuang, Hebei, China; The Key Laboratory of New Drug Pharmacology and Toxicology, Hebei Province, Shijiazhuang, Hebei, China; Department of Biopharmacy, Hebei Medical University, Shijiazhuang, Hebei, ChinaDepartment of Pharmacology, Hebei Medical University, The Key Laboratory of Neural and Vascular Biology, Ministry of Education, Shijiazhuang, Hebei, China; Department of Pharmacology, School of Pharmacy, Qingdao University, Qingdao, Shandong, ChinaDepartment of Pharmacology, Hebei Medical University, The Key Laboratory of Neural and Vascular Biology, Ministry of Education, Shijiazhuang, Hebei, ChinaDepartment of Pharmacology, Hebei Medical University, The Key Laboratory of Neural and Vascular Biology, Ministry of Education, Shijiazhuang, Hebei, ChinaDepartment of Pharmacology, Hebei Medical University, The Key Laboratory of Neural and Vascular Biology, Ministry of Education, Shijiazhuang, Hebei, China; The Key Laboratory of New Drug Pharmacology and Toxicology, Hebei Province, Shijiazhuang, Hebei, China; Corresponding author. Department of Pharmacology, Hebei Medical University, Shijiazhuang, 050017, China.The volume-regulated anion channel (VRAC) plays a central role in maintaining cell volume in response to osmotic stress. Leucine-rich repeat-containing 8A (LRRC8A) was recently identified as an essential component of VRAC although other Cl− channels were also suggested to contribute to VRAC. VRAC is activated when a cell is challenged with a hypotonic environment or even in isotonic conditions challenged with different stimuli. It is not clear how VRAC is activated and whether activation of VRAC in hypotonic and isotonic conditions share the same mechanism. In this present study, we investigated relative contribution of LRRC8A and anoctamin 1(ANO1) to VRAC currents activated by fetal bovine serum (FBS) in isotonic condition, and studied the role of intracellular Ca2+ in this activation. We used CRISPR/Cas9 gene editing approach, electrophysiology, and pharmacology approaches to show that VRAC currents induced by FBS is mostly mediated by LRRC8A in HEK293 cells, but also with significant contribution from ANO1. FBS induces Ca2+ transients and these Ca2+ signals are required for the activation of VRAC by serum. These findings will help to further understand the mechanism in activation of VRAC.http://www.sciencedirect.com/science/article/pii/S1347861320300384VRACSerumANO1LRRC8ACa2+ |
| spellingShingle | Huiran Zhang Yani Liu Hongchao Men Fan Zhang Hailin Zhang LRRCA8A and ANO1 contribute to serum-induced VRAC in a Ca2+-dependent manners Journal of Pharmacological Sciences VRAC Serum ANO1 LRRC8A Ca2+ |
| title | LRRCA8A and ANO1 contribute to serum-induced VRAC in a Ca2+-dependent manners |
| title_full | LRRCA8A and ANO1 contribute to serum-induced VRAC in a Ca2+-dependent manners |
| title_fullStr | LRRCA8A and ANO1 contribute to serum-induced VRAC in a Ca2+-dependent manners |
| title_full_unstemmed | LRRCA8A and ANO1 contribute to serum-induced VRAC in a Ca2+-dependent manners |
| title_short | LRRCA8A and ANO1 contribute to serum-induced VRAC in a Ca2+-dependent manners |
| title_sort | lrrca8a and ano1 contribute to serum induced vrac in a ca2 dependent manners |
| topic | VRAC Serum ANO1 LRRC8A Ca2+ |
| url | http://www.sciencedirect.com/science/article/pii/S1347861320300384 |
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