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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

Article DOI: 10.3791/53035-v • 08:46 min • September 1st, 2015
September 1st, 2015 •
1, 1, 1, 1, 2, 1
1Department of Physiology, Medical College of Wisconsin, 2Department of Integrative Biology & Pharmacology, University of Texas Health Science Center at Houston

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résumé
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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.

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Patch Clamp Live Fluorescence Microscopy Functional Properties Freshly Isolated PKD Epithelium Cyst Initiation Polycystic Kidney Disease Tubular Cell Proliferation Luminal Fluid Accumulation Extracellular Matrix Formation Ion Channels Intracellular Calcium Signaling Real-time Observation Patch-clamp Technique Intracellular Ca2+ Level Epithelial Monolayers Renal Cysts PCK Rats Autosomal Recessive Polycystic Kidney Disease (ARPKD) Ex Vivo Analysis Non-dilated Tubules Single Channel Activity Intracellular Ca2+ Dynamics Enzymatic Processing Native Setting High Resolution Images Antibodies Staining Dyes Preparation
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