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Implementing Patch Clamp and Live Fluorescence Microscopy to Monitor Functional Properties of Freshly Isolated PKD Epithelium
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JoVE Journal Biology
Implementing Patch Clamp and Live Fluorescence Microscopy to Monitor Functional Properties of Freshly Isolated PKD Epithelium

Implementing Patch Clamp and Live Fluorescence Microscopy to Monitor Functional Properties of Freshly Isolated PKD Epithelium

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

Ion channels expressed in renal tubular epithelium play a significant role in the pathology of polycystic kidney disease. Here we describe experimental protocols used to perform patch-clamp analysis and intracellular calcium level measurements in cystic epithelium freshly isolated from rodent kidneys.

Tags

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

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