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Arterial smooth muscle cells express segment a‐deficient TMEM16A channels (1077.7)
Author(s) -
Burris Sarah,
Jangsangthong Wanchana,
Leo M. Dennis,
Narayanan Damodaran,
Jaggar Jonathan H.
Publication year - 2014
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.28.1_supplement.1077.7
Subject(s) - patch clamp , alternative splicing , microbiology and biotechnology , exon , splice , transmembrane protein , biology , electrophysiology , contractility , chemistry , gene , endocrinology , biochemistry , neuroscience , receptor
Transmembrane protein A (TMEM16A) Cl‐ channels are expressed in arterial smooth muscle cells (SMCs) where they regulate physiological functions, including contractility and proliferation. TMEM16A alternative splice variation, which can occur at segments a, b, c, and d can alter channel biophysical properties. Identifying TMEM16A splice variants expressed in arterial SMCs should better define mechanisms by which these channels control vascular function. Our data, obtained using RT‐PCR, indicate that arterial SMCs express TMEM16A channels deficient in segment a, which removes 116 amino acids of the proximal N‐terminus. In contrast, arterial SMC TMEM16A channels contained segments b (exon 6b), c (exon13), and d (exon 15). Western blotting and whole‐cell patch clamp electrophysiology experiments demonstrated that vectors encoding segment a‐deficient TMEM16A channels (bcd) produced less protein and generated smaller whole‐cell currents than those expressing full‐length (abcd) channels in HEK293 cells. Lower TMEM16Abcd protein and currents did not occur due to attenuated transcription, elevated protein degradation or diminished surface trafficking. We are performing further experiments to test the hypothesis that segment a‐deficiency inhibits TMEM16A translation in arterial smooth muscle cells. NIH/NHLBI Grant Funding Source : NIH/NHLBI