Patch cramming: monitoring intracellular messengers in intact cells with membrane patches containing detector ion channels.
This paper introduces "patch cramming," a new procedure that utilizes an ion channel gated directly by an intracellular messenger molecule as a probe for detecting changes in the concentration of that molecule in an intact cell. A patch pipette containing the channel in a membrane patch is inserted into a recipient cell where the channel locally "senses" the intracellular messenger. In this study patches containing Ca2(+)-dependent K+ channels were inserted into Helix neurons, where they were activated by Ca2+ influx during trains of action potentials. Channels gated directly by other messengers, including cyclic nucleotides and IP3, have also been identified. Hence, by using detector channels with appropriate specificity, it may be possible to detect local intracellular fluctuations of these molecules.
Graduate Department of Biochemistry, Brandeis University, Waltham, Massachusetts 02254.
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RH
Kramer
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002462:Cell Membrane / Q000502:physiology*; D004594:Electrophysiology / Q000295:instrumentation / Q000379:methods*; D005724:Ganglia / Q000502:physiology*; D006372:Helix, Snails; D066298:In Vitro Techniques; D007473:Ion Channels / Q000502:physiology*; D015221:Potassium Channels / Q000187:drug effects / Q000502:physiology*
D007473:Ion Channels; D015221:Potassium Channels; D002118:Calcium