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Isolating and Imaging Live, Intact Pacemaker Regions of Mouse Renal Pelvis by Vibratome Sectioning.

Authors
  • Grainger, Nathan1
  • Sanders, Kenton M2
  • Drumm, Bernard T3
  • 1 Department of Physiology & Cell Biology, University of Nevada, Reno School of Medicine; Department of Physiology & Membrane Biology, University of California School of Medicine.
  • 2 Department of Physiology & Cell Biology, University of Nevada, Reno School of Medicine.
  • 3 Department of Physiology & Cell Biology, University of Nevada, Reno School of Medicine; Department of Life & Health Sciences, Dundalk Institute of Technology; [email protected]
Type
Published Article
Journal
Journal of Visualized Experiments
Publisher
MyJoVE Corporation
Publication Date
Apr 30, 2021
Issue
170
Identifiers
DOI: 10.3791/62040
PMID: 33999021
Source
Medline
Language
English
License
Unknown

Abstract

The renal pelvis (RP) is a funnel-shaped, smooth muscle structure that facilitates normal urine transport from the kidney to the ureter by regular, propulsive contractions. Regular RP contractions rely on pacemaker activity, which originates from the most proximal region of the RP at the pelvis-kidney junction (PKJ). Due to the difficulty in accessing and preserving intact preparations of the PKJ, most investigations on RP pacemaking have focused on single-cell electrophysiology and Ca2+ imaging experiments. Although important revelations on RP pacemaking have emerged from such work, these experiments have several intrinsic limitations, including the inability to accurately determine cellular identity in mixed suspensions and the lack of in situ imaging of RP pacemaker activity. These factors have resulted in a limited understanding of the mechanisms that underlie normal rhythmic RP contractions. In this paper, a protocol is described to prepare intact segments of mouse PKJ using a vibratome sectioning technique. By combining this approach with mice expressing cell-specific reporters and genetically encoded Ca2+ indicators, investigators may be able to more accurately study the specific cell types and mechanisms responsible for peristaltic RP contractions that are vital for normal urine transport.

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