Ion Channels by Bernardo Rudy (Eds.)

By Bernardo Rudy (Eds.)

This booklet describes sleek electrophysiological innovations for recording and research of ion channels in a variety of platforms together with cells in tradition, Xenopus oocytes, mind slices, mobile organelles, and microorganisms. Many chapters take care of the prevention and correction of electrophysiological artifacts. additionally integrated are tools for the purification and expression of ion channel genes and proteins

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1] PATCHCLAMPTECHNIQUES 9 25 mvl~ S ms 500 pA [ 5 ms Fie. 3. Series of superimposed current records from whole-cell recording in response to depolarizing stimuli to -20, - 1 0 , 0, and + 10 mV from a holding potential o f - 7 0 mV, using a bovine chromalfin cell at room temperature. 2 M~). phodiesterase, and cyclic GMP was present in the excised patch, in addition to the sodium channel present in the membrane. Whole-Cell Recording Following initial seal formation, it is also possible to rupture deliberately the patch of membrane trapped within the pipette by application of strong suction or a voltage pulse of several hundred millivolts.

Lindau and E. Neher, PfluegersArch. 411, 137 (1988). 14 ELECTROPHYSlOLOGICAL TECHNIQUES [2] dent calcium channels can be activated following the binding o f a ligand to receptors on the cell surface. Furthermore, both receptor-linked GTPbinding proteins and second-messenger systems within the cytosol can affect the activity o f ion channels. Thus, complex regulatory pathways linking surface receptors, the metabolism of the cell and ion channels can result in long-lasting changes in the behavior of the cell.

P. 3. Plenum, New York and London, 1983. ] PATCH CLAMP SETUP 29 Vo, which is proportional to the input current, that is, Vo = -iRf. When used in a patch voltage clamp, the patch pipette is attached to the inverting ( - ) input of the operational amplifier, and the circuit is modified so that command voltages, Vc, are applied to the noninverting (+) input; as a result of feedback the command voltage will also be imposed on the inverting input, that is, at the top of the patch pipette. It should be noted that most commercial patch damps utilize a discrete JFET input stage in conjunction with a commercial operational amplifier to create the headstage amplifier.

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