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Implementering Patch Clamp og live Fluorescens mikroskopi til Monitor funktionelle egenskaber af frisk isolerede PKD epitel
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JoVE Journal Biology
Implementing Patch Clamp and Live Fluorescence Microscopy to Monitor Functional Properties of Freshly Isolated PKD Epithelium

Implementering Patch Clamp og live Fluorescens mikroskopi til Monitor funktionelle egenskaber af frisk isolerede PKD epitel

DOI:
10.3791/53035-v

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08:46 min

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September 01, 2015

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, , , , ,
1Department of Physiology,Medical College of Wisconsin, 2Department of Integrative Biology & Pharmacology,University of Texas Health Science Center at Houston

Chapters

  • 00:05Title
  • 02:14Isolation of Renal Cysts and Connecting Tubules/Collecting Duct Segments
  • 05:02Single Channel Patch-clamp Electrophysiology
  • 05:32Ratiometric Epifluorescence Measurement of Intracellular Calcium Concentration in the Epithelial Cells
  • 06:55Results: Effect of Benzamil Treatment and ATP in the Cystic Cells of PCK Rats
  • 08:08Conclusion

Summary

Automatic Translation

Automatic Translation

Ionkanaler udtrykt i tubulær epitel spiller en væsentlig rolle i patologien af ​​polycystisk nyresygdom. Her beskriver vi forsøgsprotokoller bruges til at udføre patch-clamp analyse og intracellulært calcium niveau målinger i cystisk epitel frisk isoleret fra gnaver nyrerne.

Tags

Patch ClampLive Fluorescence MicroscopyPolycystic Kidney DiseasePKDARPKDIon ChannelsCalcium SignalingEpithelial MonolayerEx Vivo AnalysisPCK RatSprague Dawley RatNative SettingEpithelial IsolationImmunostainingPrimary CultureBiochemical Assays

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