Nuclear patch-clamp recording from inositol 1,4,5-trisphosphate receptors

Methods Cell Biol. 2010;99:199-224. doi: 10.1016/B978-0-12-374841-6.00008-6.

Abstract

Inositol 1,4,5-trisphosphate receptors (IP(3)R) are ubiquitous intracellular Ca(2+) channels. They are regulated by IP(3) and Ca(2+) and can thereby both initiate local Ca(2+) release events and regeneratively propagate Ca(2+) signals evoked by receptors that stimulate IP(3) formation. Local signaling by small numbers of IP(3)R underpins the utility of IP(3)-evoked Ca(2+) signals as a ubiquitous signaling pathway. The physiological impact of Ca(2+) release by very small numbers of IP(3)R underscores the necessity to understand the behavior of IP(3)R at the single-channel level. In addition, and in common with analyses of every other ion channel, single-channel analyses have the potential to define the steps linking IP(3) binding to channel opening. Patch-clamp recording, by resolving the openings and closings of single channels with exquisite temporal resolution, is the most powerful technique for analysis of single-channel events. It has contributed enormously to the understanding of gating and desensitization/inactivation of numerous ion channels. However, most IP(3)R reside within intracellular membranes, where they are inaccessible to conventional patch-clamp recording methods. Here, we describe the application of nuclear patch-clamp methods to single-channel analyses of native and recombinant IP(3)R.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cations
  • Cell Line
  • Cell Nucleus / metabolism*
  • Chickens
  • Gene Knockout Techniques
  • Inositol 1,4,5-Trisphosphate Receptors / metabolism*
  • Ion Channel Gating
  • Patch-Clamp Techniques / methods*

Substances

  • Cations
  • Inositol 1,4,5-Trisphosphate Receptors