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

Junqiu Yang; Kelli Delaloye; Urvi S. Lee; Jianmin Cui

doi:10.3791/2269

Journal

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Biology

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Grampo patch e Técnicas de Perfusão para estudar canais iônicos Expresso em Xenopus Oócitos

JoVE Journal
Biology

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JoVE Journal Biology

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

Grampo patch e Técnicas de Perfusão para estudar canais iônicos Expresso em Xenopus Oócitos

DOI:

10.3791/2269-v

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10:19 min

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January 10, 2011

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Junqiu Yang, Kelli Delaloye, Urvi S. Lee, Jianmin Cui

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

Chapters

00:05Title
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

Automatic Translation

Corrente iônica de canais BK é gravado usando técnicas de patch clamp. Canais BK são expressos em Oócitos Xenopus Através da injeção de RNA mensageiro. A solução intracelular durante gravações de patch clamp é controlado por um sistema de perfusão.