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A method for making solution changes in the sub-millisecond range at the tip of a patch pipette

  • Excitable Tissues and Central Nervous Physiology
  • Instruments and Techniques
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Abstract

A method is described here for making multiple fast external solution changes at the tip of a patch pipette. The time for the change, 0.2 ms, has been established by measuring changes in liquid junction potential at the tip of an open patch pipette. This technique of producing an abrupt change in solution allows agonist/receptor reactions to be studied under non-equilibrium conditions. We have applied this technique to the nicotinic receptors in outside-out patches from skeletal muscle cell line C2 (Jaffe and Saxel 1979) and from bovine adrenal chromaffin cells. The application of step changes in acetylcholine concentration produces current traces with a characteristic shape, which may be compared with the predictions of established models for the activation and desensitisation of the nicotinic receptor. The results of making single steps and also short pulses in acetylcholine concentration are demonstrated. The direct comparison of two different cholinergic agonists is demonstrated.

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References

  • Akaike N, Inoue M, Krishtal OA (1986) “Concentration clamp” study of gamma-aminobutyric-acid-induced chloride current kinetics in frog sensory neurones. J Physiol 379:171–185

    Google Scholar 

  • Brett RS, Dilger JP, Adams PR, Lancaster B (1986) A method for the rapid exchange of solutions bathing excised membrane patches. Biophys J 50:987–991

    Google Scholar 

  • Boll W, Lux HD (1985) Action of organic antagonists on neuronal calcium currents. Neurosci Lett 56:335–339

    Google Scholar 

  • Colquhoun D, Hawkes AG (1977) Relaxation and fluctuation of membrane currents that flow through drug operated channels. Proc R Soc Lond B 199:15–52

    Google Scholar 

  • Colquhoun D, Sakmann B (1985) Fast events in single-channel currents activated by acetylcholine and its analogues at the frog muscle endplate. J Physiol 369:501–557

    Google Scholar 

  • Dudel J, Franke CH, Hatt H (1988) Glutamate activated membrane channels in crustacean muscle fibres. Neurotoxicology 88:405–418

    Google Scholar 

  • Dudel J, Franke C, Hatt H, Ramsey RL, Usherwood PNR (1988) Rapid activation and desensitisation by glutamate of excitatory, cation selective channels in locust muscle. Neurosci Lett 88:33–38

    Google Scholar 

  • Franke CH, Hatt H, Dudel J (1987) Liquid filament switch for ultra-fast exchanges of solutions at excised patches of synaptic membrane of crayfish muscle. Neurosci Lett 77:199–204

    Google Scholar 

  • Hamill OP, Marty A, Neher E, Sakmann B, Sigworth FJ (1981) Improved patch-clamp techniques for high resolution current recording from cells and cell free membranes. Pflügers Arch 391:85–100

    Google Scholar 

  • Hattori K, Akaike OA, Oomura Y, Kuraoka S (1984) Internal perfusion studies demonstrating GABA-induced chloride responses in frog primary afferent neurones. Am J Physiol 246:C259–265

    Google Scholar 

  • Hodgkin AL, Horowicz P (1960) The effect of sudden changes of ionic concentration on the membrane of single muscle fibres. J Physiol Lond 153:370–385

    Google Scholar 

  • Huck S, Lux HD (1987) Patch-clamp study of ion channels activated by GABA and glycine in cultured cerebellar neurones of the mouse. Neurosci Lett 79(1–2):103–107

    Google Scholar 

  • Jaffe D, Saxel O (1979) Serial passaging and differentiation of myogenic cells isolated from dystrophic mouse muscle. Nature 270:725–727

    Google Scholar 

  • Kakei M, Ashcroft FM (1987) A microflow superfusion system for use with excised patches. Pflügers Arch 409:337–341

    Google Scholar 

  • Knight DE, Baker PF (1983) Stimulus secretion coupling in isolated bovine adrenal medullary cells. Q J Exp Physiol 68:123–143

    Google Scholar 

  • Konnerth A, Lux HD, Morad M (1987) Proton-induced transformation of calcium channel in chick dorsal root ganglion cells. J Physiol (Lond) 386:571–601

    Google Scholar 

  • Kostyuk PG, Krishtal OA, Pidoplichko VI (1981) Intracellular perfusion. J Neurosci Methods 4(3):201–210

    Google Scholar 

  • Krishtal OA, Marchenko SM, Pidoplichko VI (1983) Receptor for ATP in the membrane of mammalian sensory neurones. Neurosci Lett 35:42–45

    Google Scholar 

  • Maconochie DJ (1988) Solution changes in 0.2 ms at the tip of a patch pipette. Proc Physiol Soc 399:8P

  • Ogden DC, Colquhoun D (1985) Ion channel block by acetylcholine, carbachol and subderylcholine at the frog neuromuscular junction. Proc R Soc Lond [Biol] 225:329–355

    Google Scholar 

  • Olesen S-P, Clapham DE, Davies PF (1988) Haemodynamic shear stress activates a potassium current in vascular endothelial cells. Nature 331:168–170

    Google Scholar 

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Maconochie, D.J., Knight, D.E. A method for making solution changes in the sub-millisecond range at the tip of a patch pipette. Pflugers Arch. 414, 589–596 (1989). https://doi.org/10.1007/BF00580996

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  • DOI: https://doi.org/10.1007/BF00580996

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