A nonenzymatic preparation of epithelial basolateral membrane for patch clamp

F. Wehner, L. Garretson, K. Dawson, Y. Segal, L. Reuss

Research output: Contribution to journalArticlepeer-review

5 Scopus citations

Abstract

A preparation has been developed that permits patch clamping of the basolateral membrane of Necturus gallbladder epithelial cells with a high success rate. The epithelium is separated from the underlying tissues mechanically, without enzymatic treatment. Its apical surface is attached to a plastic cover slip, and the basolateral surface, facing up, is cleaned with a suction pipette under microscopic observation. With this cleaning procedure, the success rate in obtaining gigaohm seals increases from < 1% to ~ 10% of the attempts. The cells appear to retain their structural and functional integrity, as evidenced by electron-microscopic appearance and magnitude of cell membrane voltages. Major advantages of the preparation are that the basolateral membrane domain is preserved and that enzymatic treatment, which could potentially alter membrane proteins, is not necessary.

Original languageEnglish (US)
Pages (from-to)C1159-C1164
JournalAmerican Journal of Physiology - Cell Physiology
Volume258
Issue number6 27-6
DOIs
StatePublished - 1990

Keywords

  • electron microscopy
  • ion channels
  • potassium channel

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