PUBLICATION

Role of zebrafish ClC-K/barttin channels in apical kidney chloride reabsorption

Authors
Pérez-Rius, C., Castellanos, A., Gaitán-Peñas, H., Navarro, A., Artuch, R., Barrallo-Gimeno, A., Estévez, R.
ID
ZDB-PUB-190610-4
Date
2019
Source
The Journal of physiology   597(15): 3969-3983 (Journal)
Registered Authors
Keywords
none
MeSH Terms
  • Animals
  • Chlorides/metabolism
  • HEK293 Cells
  • Humans
  • Kidney/metabolism*
  • Mutation
  • Protein Transport
  • Renal Reabsorption*
  • Zebrafish
(all 9)
PubMed
31177533 Full text @ J. Physiol.
Abstract
Key points We have characterized the zebrafish clc-k and barttin proteins, demonstrating that they form a protein complex mediating chloride flux in a similar manner to their mammalian counterparts. As in mammals, in zebrafish, clc-k and barttin are basically expressed in the kidney. Contrary to what is found in mammals, in zebrafish both proteins show an apical localization in the kidney. We have generated the first knockout in zebrafish of a CLC protein. Lack of clc-k in zebrafish resulted in embryonic lethality, possibly caused by a reduction in total chloride content. As a consequence, there is an up-regulation other chloride channels and other regulatory mechanism such as renin or the uro-guanylin receptor in the kidney. barttin is mislocalized in vivo when clc-k is not present, indicating that there is a mutual dependence of the protein expression and localization between barttin and clc-k proteins.
Abstract ClC-K/barttin channels are very important for salt transport in the kidney. This function can be clearly seen as mutations in CLCNKB or BSND cause Bartter's syndrome type III and IV, respectively. Working with the freshwater teleost zebrafish, we characterized the genes homologous to the mammalian chloride channel ClC-K and its obligate subunit barttin and we obtained and studied clc-k knockout zebrafish. The zebrafish clc-k/barttin proteins are very similar to their mammalian counterparts, and both proteins are necessary to mediate chloride currents. Localization studies indicated that both proteins are exclusively expressed in the apical membranes of zebrafish kidney tubules. Knockout of clc-k resulted in embryonic lethality. These animals showed barttin mislocalization and a reduction in whole-body chloride concentration, as well as up-regulation of the expression of other chloride channels, renin and also an increase in the kidney expression of the uroguanylin receptor. Our results indicate that apical kidney chloride reabsorption through clc-k/barttin channels is crucial for chloride homeostasis in zebrafish as happens in humans. The zebrafish model could be used as a new in vivo system to study ClC-K function. This article is protected by copyright. All rights reserved.
Genes / Markers
Marker Marker Type Name
bsndGENEbarttin CLCNK type accessory subunit beta
cftrGENECF transmembrane conductance regulator
clcn2aGENEchloride channel, voltage-sensitive 2a
clcn2bGENEchloride channel, voltage-sensitive 2b
clcn2cGENEchloride channel 2c
clcnkGENEchloride channel K
dhcr24GENE24-dehydrocholesterol reductase
dsc2lGENEdesmocollin 2 like
dsg2.1GENEdesmoglein 2, tandem duplicate 1
epha2bGENEeph receptor A2 b
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Figures
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Expression
Phenotype
Fish Conditions Stage Phenotype Figure
WThypotonicDay 5
WThypertonicDay 5
clcnksa18332/sa18332standard conditionsDays 7-13
clcnksa18332/sa18332standard conditionsDays 7-13
clcnksa18332/sa18332standard conditionsDays 7-13
clcnksa18332/sa18332standard conditionsDays 7-13
clcnksa18332/sa18332standard conditionsDays 7-13
clcnksa18332/sa18332standard conditionsDays 7-13
clcnksa18332/sa18332standard conditionsDays 7-13
clcnksa18332/sa18332standard conditionsDays 7-13
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Mutations / Transgenics
Allele Construct Type Affected Genomic Region
nkgSAlzGFFD875BGtTransgenic Insertion
nkuasgfp1aTgTransgenic Insertion
    sa18332
      		Point Mutation
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      Human Disease / Model
      No data available
      Sequence Targeting Reagents
      No data available
      Fish
      Fish
      clcnksa18332/sa18332
      clcnksa18332/sa18332; gucy2cankgSAlzGFFD875BGt; nkuasgfp1aTg
      WT
      1 - 3 of 3
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      Antibodies
      Orthology
      Engineered Foreign Genes
      Marker Marker Type Name
      EGFPEFGEGFP
      GAL4FFEFGGAL4FF
      1 - 2 of 2
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      Mapping
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      Citation
      Pérez-Rius, C., Castellanos, A., Gaitán-Peñas, H., Navarro, A., Artuch, R., Barrallo-Gimeno, A., Estévez, R. (2019) Role of zebrafish ClC-K/barttin channels in apical kidney chloride reabsorption. The Journal of physiology. 597(15):3969-3983.