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Renal epithelial electrophysiological response to cobalt chloride induced hypoxia (888.4)
Author(s) -
Nag Subhra,
Resnick Andrew
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.888.4
Subject(s) - epithelial sodium channel , hypoxia (environmental) , sodium , amiloride , chemistry , endocrinology , medicine , polycystic kidney disease , viability assay , kidney , microbiology and biotechnology , biology , cell , biochemistry , oxygen , organic chemistry
Regulation of amiloride‐sensitive sodium current i.e., sodium channel (ENaC) current was found to be dependent on intact cilium in a kidney epithelial cell culture model. We hypothesize that cystic disease, for example Polycystic Kidney Disease (PKD), results in localized hypoxia due to creation of stagnant fluid‐filled cysts, interfering with normal salt and water transport. In the current study, we used a cortical collecting duct cell line (mCCD 1296 (d)) of an Immortomouse. We simulated hypoxic conditions with 100 µM Cobalt Chloride (CoCl2) and observed sodium transport under static and dynamic cell differentiation conditions. CoCl2 simulates hypoxia conditions by stabilizing the hypoxia inducible transcription factor 1α (HIF1α). We noticed a drop in the sodium current at time points of 24, 48, 72, and 96 hours in the presence of CoCl2. According to our hypothesis, the sodium current is expected to decrease in shorter time points of hypoxia whereas longer hypoxia induces cellular apoptosis totally nullifying the sodium current. We record sodium current in response to hypoxia at shorter time intervals within 48 hours as well as longer time periods upto 8 days. We use MTT cell viability assay to determine the time points at which CoCl2 induced hypoxia or cytotoxic CoCl2 start causing cell death. Correlating the MTT assay results with sodium current results will help us finding the direct correlation between hypoxia and sodium transport. This electrophysiological study will be complimented with immunocytochemistry to image cilium length alterations, and ciliary‐associated proteins, STAT‐6 and Phosphorylated‐STAT6 localizations. Thus, this study will provide better understanding of how hypoxia, sodium ion transport, effect of fluid flow and physiological and molecular cellular responses are interrelated. Grant Funding Source : Supported by the National Institute of Diabetes and Digestive and Kidney Diseases 1R15DK092716