Alteration of active Na-K transport on protein kinase C activation in cultured ciliary epithelium

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Abstract

Purpose. Experiments were conducted to test whether protein kinase C activation causes changes in active sodium-potassium transport in an established SV-10 transformed line (ODM2) of cultured human nonpigmented ciliary epithelial cells. Methods. Rubidium-86 (86Rb) uptake was measured and the data used to determine the rate of potassium entry into the cells. Results. Protein kinase C activator, phorbol dibutyrate (PDBu), caused a stimulation of ouabain-sensitive 86Rb uptake. Inhibition of protein kinase C by 1-(5-isoquinolinylsulfonyl) 2-methylpiperazine (H-7), or down-regulation of protein kinase C activation by prolonged exposure of PDBu, decreased the PDBu response. These results suggest that protein kinase C plays a role in Na-K pump activation. The Na+/H+ exchanger inhibitor, amiloride, also reduced the stimulation of the ouabain-sensitive 86Rb uptake by PDBu. 86Rb efflux was not altered by protein kinase C activation. At the same time that PDBu increased the ouabain-sensitive 86Rb uptake, it also decreased the ouabain-insensitive 86Rb uptake. The ouabain-insensitive 86Rb uptake component could be inhibited by bumetanide, suggesting that protein kinase C activation decreases the activity of a Na/K/2Cl cotransporter. Conclusions. These findings suggest that activation of protein kinase C may stimulate Na,K-ATPase activity mainly by a mechanism involving increased Na+ influx mediated by the Na+/H+ exchanger.

Original languageEnglish (US)
Pages (from-to)539-546
Number of pages8
JournalInvestigative Ophthalmology and Visual Science
Volume34
Issue number3
StatePublished - 1993
Externally publishedYes

Fingerprint

Protein Kinase C
Carrier Proteins
Epithelium
Ouabain
1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine
Sodium-Hydrogen Antiporter
Potassium
Rubidium
Bumetanide
Amiloride
Down-Regulation
Epithelial Cells
Sodium
phorbol

Keywords

  • Rb uptake
  • bumetanide
  • human ciliary epithelium
  • Na,K-ATPase
  • phorbol dibutyrate
  • protein kinase C

ASJC Scopus subject areas

  • Ophthalmology

Cite this

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title = "Alteration of active Na-K transport on protein kinase C activation in cultured ciliary epithelium",
abstract = "Purpose. Experiments were conducted to test whether protein kinase C activation causes changes in active sodium-potassium transport in an established SV-10 transformed line (ODM2) of cultured human nonpigmented ciliary epithelial cells. Methods. Rubidium-86 (86Rb) uptake was measured and the data used to determine the rate of potassium entry into the cells. Results. Protein kinase C activator, phorbol dibutyrate (PDBu), caused a stimulation of ouabain-sensitive 86Rb uptake. Inhibition of protein kinase C by 1-(5-isoquinolinylsulfonyl) 2-methylpiperazine (H-7), or down-regulation of protein kinase C activation by prolonged exposure of PDBu, decreased the PDBu response. These results suggest that protein kinase C plays a role in Na-K pump activation. The Na+/H+ exchanger inhibitor, amiloride, also reduced the stimulation of the ouabain-sensitive 86Rb uptake by PDBu. 86Rb efflux was not altered by protein kinase C activation. At the same time that PDBu increased the ouabain-sensitive 86Rb uptake, it also decreased the ouabain-insensitive 86Rb uptake. The ouabain-insensitive 86Rb uptake component could be inhibited by bumetanide, suggesting that protein kinase C activation decreases the activity of a Na/K/2Cl cotransporter. Conclusions. These findings suggest that activation of protein kinase C may stimulate Na,K-ATPase activity mainly by a mechanism involving increased Na+ influx mediated by the Na+/H+ exchanger.",
keywords = "Rb uptake, bumetanide, human ciliary epithelium, Na,K-ATPase, phorbol dibutyrate, protein kinase C",
author = "T. Mito and Delamere, {Nicholas A}",
year = "1993",
language = "English (US)",
volume = "34",
pages = "539--546",
journal = "Investigative Ophthalmology and Visual Science",
issn = "0146-0404",
publisher = "Association for Research in Vision and Ophthalmology Inc.",
number = "3",

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TY - JOUR

T1 - Alteration of active Na-K transport on protein kinase C activation in cultured ciliary epithelium

AU - Mito, T.

AU - Delamere, Nicholas A

PY - 1993

Y1 - 1993

N2 - Purpose. Experiments were conducted to test whether protein kinase C activation causes changes in active sodium-potassium transport in an established SV-10 transformed line (ODM2) of cultured human nonpigmented ciliary epithelial cells. Methods. Rubidium-86 (86Rb) uptake was measured and the data used to determine the rate of potassium entry into the cells. Results. Protein kinase C activator, phorbol dibutyrate (PDBu), caused a stimulation of ouabain-sensitive 86Rb uptake. Inhibition of protein kinase C by 1-(5-isoquinolinylsulfonyl) 2-methylpiperazine (H-7), or down-regulation of protein kinase C activation by prolonged exposure of PDBu, decreased the PDBu response. These results suggest that protein kinase C plays a role in Na-K pump activation. The Na+/H+ exchanger inhibitor, amiloride, also reduced the stimulation of the ouabain-sensitive 86Rb uptake by PDBu. 86Rb efflux was not altered by protein kinase C activation. At the same time that PDBu increased the ouabain-sensitive 86Rb uptake, it also decreased the ouabain-insensitive 86Rb uptake. The ouabain-insensitive 86Rb uptake component could be inhibited by bumetanide, suggesting that protein kinase C activation decreases the activity of a Na/K/2Cl cotransporter. Conclusions. These findings suggest that activation of protein kinase C may stimulate Na,K-ATPase activity mainly by a mechanism involving increased Na+ influx mediated by the Na+/H+ exchanger.

AB - Purpose. Experiments were conducted to test whether protein kinase C activation causes changes in active sodium-potassium transport in an established SV-10 transformed line (ODM2) of cultured human nonpigmented ciliary epithelial cells. Methods. Rubidium-86 (86Rb) uptake was measured and the data used to determine the rate of potassium entry into the cells. Results. Protein kinase C activator, phorbol dibutyrate (PDBu), caused a stimulation of ouabain-sensitive 86Rb uptake. Inhibition of protein kinase C by 1-(5-isoquinolinylsulfonyl) 2-methylpiperazine (H-7), or down-regulation of protein kinase C activation by prolonged exposure of PDBu, decreased the PDBu response. These results suggest that protein kinase C plays a role in Na-K pump activation. The Na+/H+ exchanger inhibitor, amiloride, also reduced the stimulation of the ouabain-sensitive 86Rb uptake by PDBu. 86Rb efflux was not altered by protein kinase C activation. At the same time that PDBu increased the ouabain-sensitive 86Rb uptake, it also decreased the ouabain-insensitive 86Rb uptake. The ouabain-insensitive 86Rb uptake component could be inhibited by bumetanide, suggesting that protein kinase C activation decreases the activity of a Na/K/2Cl cotransporter. Conclusions. These findings suggest that activation of protein kinase C may stimulate Na,K-ATPase activity mainly by a mechanism involving increased Na+ influx mediated by the Na+/H+ exchanger.

KW - Rb uptake

KW - bumetanide

KW - human ciliary epithelium

KW - Na,K-ATPase

KW - phorbol dibutyrate

KW - protein kinase C

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