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The Calcium‐Activated Chloride Channel, Anoctamin‐1, is a Critical Regulator of Cell Proliferation Mediated via p44\42 MAPK Phosphorylation and Calcium Signaling
Author(s) -
Franklin Brandon M,
Osborn Jeffrey L
Publication year - 2016
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.30.1_supplement.1224.45
Subject(s) - microbiology and biotechnology , dids , chloride channel , chemistry , mapk/erk pathway , cell growth , calcium , calcium in biology , signal transduction , biology , biochemistry , intracellular , organic chemistry , membrane
Soft tissue repair is a critical function of living organisms which requires restoration of anatomical structure and function following tissue damage. A better understanding of the cellular and molecular mechanisms that control critical cellular processes, such as proliferation, is crucial to the development of clinical therapies in regenerative medicine. We have recently shown that chloride channel inhibition, via 4,4′‐Diisothiocyano‐2,2′‐stilbenedisulfonic acid (DIDS), 4,4′‐Dinitro‐stilbene‐2,2′‐disulfonic acid (DNDS), T16a(inh)‐A01 or Cl − free environment effectively inhibits or delays tail regeneration in larval Ambystoma mexicanum salamanders. Proliferation was assessed via EdU incorporation assay, which revealed that this effect was manifested through regulation of mesenchymal cell proliferation, with all 3 pharmacological treatments reducing proliferation rate in the mesenchyme (Control: 33.5±3.2%; DIDS: 18.13±2.25%; DNDS: 19.64±3.79%; T16a(inh)‐A01: 19.65±1.85% EdU+ cells). There is no effect on proliferation in either neural or epidermal tissues. Western blot analysis demonstrated that phosphorylation of p44/42 MAPK (Erk1/2) is reduced when the calcium activated chloride channel, Tmem16a, is inhibited with T16a(inh)‐A01 in 72 hpa regenerates. These results indicate that the calcium activated chloride channel, anoctamin‐1, regulates proliferation via p44\42 MAPK. A transgenic HEK 293 cell line that overexpresses the anoctamin‐1 channel is being used to further elucidate the channel mechanisms involved in Cl‐channel mediated cell proliferation. Fluorescent calcium indicators will provide semi‐quantitative evidence regarding the influence of anoctamin‐1 on intracellular calcium signaling. Support or Funding Information Supported by Biomedical Sciences Research Group, LLC.