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Recording Ion Channels in Isolated, Split-Opened Tubules

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Ion Channels

Part of the book series: Methods in Molecular Biology ((MIMB,volume 998))

Abstract

Ion channels play key roles in physiology. They function as protein transducers able to transform stimuli and chemical gradients into electrical signals. They also are critical for cell signaling and play a particularly important role in epithelial transport acting as gateways for the movement of electrolytes across epithelial cell membranes. Experimental limitations, though, have hampered the recording of ion channel activity in many types of tissue. This has slowed progress in understanding the cellular and physiological function of these channels with most function inferred from in vitro systems and cell culture models. In many cases, such inferences have clouded rather than clarified the picture. Here, we describe a contemporary method for isolating and patch-clamping renal tubules for ex vivo analysis of ion channel function in native tissue. Focus is placed on quantifying the activity of the epithelial Na+ channel (ENaC) in the aldosterone-­sensitive distal nephron (ASDN). This isolated, split-open tubule preparation enables recording of renal ion channels in the close-to-native environment under the control of native cell signaling pathways and receptors. When combined with complementary measurements of organ and system function, and contemporary molecular genetics and pharmacology used to manipulate function and regulation, patch-clamping renal channels in the isolated, split-open tubule enables understanding to emerge about the physiological function of these key proteins from the molecule to the whole animal.

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Acknowledgments

Research from the author’s laboratories is supported by the NIH grants R01 DK59594, R01 DK087460, R01 DK070571 (to J.D.S.), and R01HL108880 (to A.S.).

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Authors and Affiliations

  1. Department of Physiology, University of Texas Health Science Center at San Antonio, San Antonio, TX, USA

    Elena Mironova, Vladislav Bugay & James D. Stockand

  2. Department of Integrative Biology and Pharmacology, University of Texas Health Science Center at Houston, Houston, TX, USA

    Oleh Pochynyuk

  3. Department of Physiology, Medical College of Wisconsin, Milwaukee, WI, USA

    Alexander Staruschenko

Authors
  1. Elena Mironova
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  2. Vladislav Bugay
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  3. Oleh Pochynyuk
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  4. Alexander Staruschenko
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  5. James D. Stockand
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Editors and Affiliations

  1. School of Biomedical Sciences, University of Leeds, Garstang Bld. 6.31d, Leeds, LS2 9JT, United Kingdom

    Nikita Gamper

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Mironova, E., Bugay, V., Pochynyuk, O., Staruschenko, A., Stockand, J.D. (2013). Recording Ion Channels in Isolated, Split-Opened Tubules. In: Gamper, N. (eds) Ion Channels. Methods in Molecular Biology, vol 998. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-351-0_27

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  • DOI: https://doi.org/10.1007/978-1-62703-351-0_27

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-350-3

  • Online ISBN: 978-1-62703-351-0

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