Discrete control of TRPV4 channel function in the distal nephron by protein kinases a and C

Mykola Mamenko, Oleg L. Zaika, Nabila Boukelmoune, Jonathan Berrout, Roger G. O'Neil, Oleh Pochynyuk

Research output: Contribution to journalArticle

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Abstract

We have recently documented that the Ca2+-permeable TRPV4 channel, which is abundantly expressed in distal nephron cells, mediates cellular Ca2+ responses to elevated luminal flow. In this study, we combined Fura-2-based [Ca2+]i imaging with immunofluorescence microscopy in isolated split-opened distal nephrons of C57BL/6 mice to probe the molecular determinants of TRPV4 activity and subcellular distribution. We found that activation of the PKC pathway with phorbol 12-myristate 13-acetate significantly increased [Ca2+] i responses to flow without affecting the subcellular distribution of TRPV4. Inhibition of PKC with bisindolylmaleimide I diminished cellular responses to elevated flow. In contrast, activation of the PKA pathway with forskolin did not affect TRPV4-mediated [Ca2+]i responses to flow but markedly shifted the subcellular distribution of the channel toward the apical membrane. These actions were blocked with the specific PKA inhibitor H-89. Concomitant activation of the PKA and PKC cascades additively enhanced the amplitude of flow-induced [Ca2+]i responses and greatly increased basal [Ca2+]i levels, indicating constitutive TRPV4 activation. This effect was precluded by the selective TRPV4 antagonist HC-067047. Therefore, the functional status of the TRPV4 channel in the distal nephron is regulated by two distinct signaling pathways. Although the PKA-dependent cascade promotes TRPV4 trafficking and translocation to the apical membrane, the PKC-dependent pathway increases the activity of the channel on the plasma membrane.

Original languageEnglish (US)
Pages (from-to)20306-20314
Number of pages9
JournalJournal of Biological Chemistry
Volume288
Issue number28
DOIs
StatePublished - Jul 12 2013

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Nephrons
Protein Kinases
Protein Kinase C
Chemical activation
Molecular Probes
Membranes
Fura-2
Colforsin
Inbred C57BL Mouse
Fluorescence Microscopy
Acetates
Cell membranes
Cell Membrane
Microscopic examination
Imaging techniques

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

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Discrete control of TRPV4 channel function in the distal nephron by protein kinases a and C. / Mamenko, Mykola; Zaika, Oleg L.; Boukelmoune, Nabila; Berrout, Jonathan; O'Neil, Roger G.; Pochynyuk, Oleh.

In: Journal of Biological Chemistry, Vol. 288, No. 28, 12.07.2013, p. 20306-20314.

Research output: Contribution to journalArticle

Mamenko, M, Zaika, OL, Boukelmoune, N, Berrout, J, O'Neil, RG & Pochynyuk, O 2013, 'Discrete control of TRPV4 channel function in the distal nephron by protein kinases a and C', Journal of Biological Chemistry, vol. 288, no. 28, pp. 20306-20314. https://doi.org/10.1074/jbc.M113.466797
Mamenko, Mykola ; Zaika, Oleg L. ; Boukelmoune, Nabila ; Berrout, Jonathan ; O'Neil, Roger G. ; Pochynyuk, Oleh. / Discrete control of TRPV4 channel function in the distal nephron by protein kinases a and C. In: Journal of Biological Chemistry. 2013 ; Vol. 288, No. 28. pp. 20306-20314.
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