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Patch Clamp and Perfusion Techniques for Studying Ion Channels Expressed in Xenopus oocytes
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Biologia
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JoVE Journal Biologia
Patch Clamp and Perfusion Techniques for Studying Ion Channels Expressed in Xenopus oocytes

Patch Clamp and Perfusion Techniques for Studying Ion Channels Expressed in Xenopus oocytes

DOI:
10.3791/2269-v

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10:19 min

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January 10, 2011

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, , ,
1Department of Energy, Environmental & Chemical Engineering,Washington University in St. Louis, 2Department of Biomedical Engineering,Washington University in St. Louis, 3Department of Biomedical Engineering and Cardiac Bioelectricity and Arrhythmia Center,Washington University in St. Louis

Capítulos

  • 00:05Título
  • 00:45Introduction
  • 01:10Injecting mRNA into Oocytes
  • 01:51Preparing the Perfusion System
  • 04:26Preparing the Ag/AgCl Wire and the Data Acquisition Equipment
  • 05:33Stripping the Vitelline Membrane from the Oocyte
  • 06:14Preparing the Patch Pipettes
  • 07:06Obtaining an Inside-Out Patch
  • 08:08Patch Clamping
  • 08:41Recording of Wild-Type BK Channels on and Oocyte Membrane Patch
  • 09:57Conclusion

Summary

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Ionic current of BK channels is recorded using patch clamp techniques. BK channels are expressed in Xenopus oocytes by injecting messenger RNA. The intracellular solution during patch clamp recordings is controlled by a perfusion system.

Tags

Patch ClampPerfusion TechniquesIon ChannelsXenopus OocytesBK ChannelsActivationStructure-function RelationshipNeurotransmitter ReceptorsSmooth Muscle ContractionHair Cell Frequency TuningNeurotransmitter ReleaseMembrane DepolarizationIntracellular Ca2+ And Mg2+Messenger RNA InjectionIncubationInside-out ConfigurationSingle Or Multiple ChannelsRecording CurrentsControlled Voltage

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