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J Am Soc Nephrol. 2015 Nov;26(11):2731-40. doi: 10.1681/ASN.2014070705. Epub 2015 Mar 18.

Visualization of Calcium Dynamics in Kidney Proximal Tubules.

Author information

1
Institute of Enzymology, Research Centre for Natural Sciences, Hungarian Academy of Sciences, Budapest, Hungary;
2
First Department of Pediatrics.
3
Department of Biophysics and Radiation Biology, MTA-SE Molecular Biophysics Research Group, and.
4
MTA-SE, Pediatrics and Nephrology Research Group, Budapest, Hungary;
5
NARIC-ABI, Gödöllö, Hungary;
6
ImmunoGenes Kft., Budakeszi, Hungary; and.
7
Department of Experimental Pharmacology, National Institute of Oncology, Budapest, Hungary.
8
Second Department of Pathology, Semmelweis University, Budapest, Hungary;
9
Institute of Enzymology, Research Centre for Natural Sciences, Hungarian Academy of Sciences, Budapest, Hungary; apati@biomembrane.hu sarkadi@biomembrane.hu.
10
Institute of Enzymology, Research Centre for Natural Sciences, Hungarian Academy of Sciences, Budapest, Hungary; Department of Biophysics and Radiation Biology, MTA-SE Molecular Biophysics Research Group, and apati@biomembrane.hu sarkadi@biomembrane.hu.

Abstract

Intrarenal changes in cytoplasmic calcium levels have a key role in determining pathologic and pharmacologic responses in major kidney diseases. However, cell-specific delivery of calcium-sensitive probes in vivo remains problematic. We generated a transgenic rat stably expressing the green fluorescent protein-calmodulin-based genetically encoded calcium indicator (GCaMP2) predominantly in the kidney proximal tubules. The transposon-based method used allowed the generation of homozygous transgenic rats containing one copy of the transgene per allele with a defined insertion pattern, without genetic or phenotypic alterations. We applied in vitro confocal and in vivo two-photon microscopy to examine basal calcium levels and ligand- and drug-induced alterations in these levels in proximal tubular epithelial cells. Notably, renal ischemia induced a transient increase in cellular calcium, and reperfusion resulted in a secondary calcium load, which was significantly decreased by systemic administration of specific blockers of the angiotensin receptor and the Na-Ca exchanger. The parallel examination of in vivo cellular calcium dynamics and renal circulation by fluorescent probes opens new possibilities for physiologic and pharmacologic investigations.

KEYWORDS:

calcium; hypoxia; ischemia-reperfusion; renal ischemia; renal proximal tubule cell

PMID:
25788535
PMCID:
PMC4625667
DOI:
10.1681/ASN.2014070705
[Indexed for MEDLINE]
Free PMC Article

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