Intracellular Ca 2+ and PKC activation do not inhibit Na + and water transport in rat CCD

Alexander Rouch, L. Chen, L. H. Kudo, P. D. Bell, B. C. Fowler, B. D. Corbitt, J. A. Schafer

Research output: Contribution to journalArticle

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Abstract

Experiments examined the effects of elevation of intracellular calcium concentration ([Ca 2+ ](i)) or activation of protein kinase C (PKC) on Na + and water transport in the rat cortical collecting duct (CCD). We measured the lumen-to-bath 22 Na + flux (J(l→b)), transepithelial voltage (V(T)), and water permeability (P(f)) in CCD from deoxycorticosterone (DOC)-treated rats. Ionomycin (0.5 and 1 μM) and thapsigargin (1 and 2 μM) were used to increase [Ca 2+ ](i). Phorbol 12-myristate 13-acetate (PMA; 0.3 and 1 μM) and oleoyl-acetyl-glycerol (OAG; 100 μM) were used as activators of PKC. [Ca 2+ ](i) was measured in isolated perfused tubules using the fluorescent dye fura 2. When added to the bathing solution, 220 pM arginine vasopressin (AVP) failed to affect [Ca 2+ ](i), whereas 1 μM ionomycin increased [Ca 2+ ](i) by 103 ± 15% and 2 μM thapsigargin increased [Ca 2+ ](i) by 24 ± 4%. In flux studies, neither ionomycin nor thapsigargin affected J(l→b) or P(f), although ionomycin caused marked morphological changes. Ionomycin also failed to alter either parameter in tubules from non-DOC-treated rats. Neither 100 μM OAG nor 1 μM PMA affected J(l→b) or P(f). OAG at 50 μM had no effect on V(T) or transepithelial resistance, indicating no inhibition of conductive Na + transport. We conclude that increased [Ca 2+ ](i) and PKC activation do not affect J(l→b) or P(f) in the rat CCD. These findings may account for the sustained increase in J(l→b) produced in the rat CCD by AVP.

Original languageEnglish
JournalAmerican Journal of Physiology - Renal Fluid and Electrolyte Physiology
Volume265
Issue number4 34-4
StatePublished - 1 Jan 1993

Fingerprint

Ionomycin
Protein Kinase C
Thapsigargin
Water
Arginine Vasopressin
Desoxycorticosterone
Fura-2
Baths
Fluorescent Dyes
Glycerol
Permeability
Acetates
Calcium

Keywords

  • arginine vasopressin
  • cortical collecting duct
  • intracellular second messengers
  • ionomycin
  • mineralocorticoid
  • oleoyl-acetyl-glycerol
  • phorbol 12-myristate 13- acetate
  • protein kinase C
  • sodium channel
  • thapsigargin
  • vasopressin

Cite this

Rouch, Alexander ; Chen, L. ; Kudo, L. H. ; Bell, P. D. ; Fowler, B. C. ; Corbitt, B. D. ; Schafer, J. A. / Intracellular Ca 2+ and PKC activation do not inhibit Na + and water transport in rat CCD In: American Journal of Physiology - Renal Fluid and Electrolyte Physiology. 1993 ; Vol. 265, No. 4 34-4.
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abstract = "Experiments examined the effects of elevation of intracellular calcium concentration ([Ca 2+ ](i)) or activation of protein kinase C (PKC) on Na + and water transport in the rat cortical collecting duct (CCD). We measured the lumen-to-bath 22 Na + flux (J(l→b)), transepithelial voltage (V(T)), and water permeability (P(f)) in CCD from deoxycorticosterone (DOC)-treated rats. Ionomycin (0.5 and 1 μM) and thapsigargin (1 and 2 μM) were used to increase [Ca 2+ ](i). Phorbol 12-myristate 13-acetate (PMA; 0.3 and 1 μM) and oleoyl-acetyl-glycerol (OAG; 100 μM) were used as activators of PKC. [Ca 2+ ](i) was measured in isolated perfused tubules using the fluorescent dye fura 2. When added to the bathing solution, 220 pM arginine vasopressin (AVP) failed to affect [Ca 2+ ](i), whereas 1 μM ionomycin increased [Ca 2+ ](i) by 103 ± 15{\%} and 2 μM thapsigargin increased [Ca 2+ ](i) by 24 ± 4{\%}. In flux studies, neither ionomycin nor thapsigargin affected J(l→b) or P(f), although ionomycin caused marked morphological changes. Ionomycin also failed to alter either parameter in tubules from non-DOC-treated rats. Neither 100 μM OAG nor 1 μM PMA affected J(l→b) or P(f). OAG at 50 μM had no effect on V(T) or transepithelial resistance, indicating no inhibition of conductive Na + transport. We conclude that increased [Ca 2+ ](i) and PKC activation do not affect J(l→b) or P(f) in the rat CCD. These findings may account for the sustained increase in J(l→b) produced in the rat CCD by AVP.",
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author = "Alexander Rouch and L. Chen and Kudo, {L. H.} and Bell, {P. D.} and Fowler, {B. C.} and Corbitt, {B. D.} and Schafer, {J. A.}",
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Intracellular Ca 2+ and PKC activation do not inhibit Na + and water transport in rat CCD . / Rouch, Alexander; Chen, L.; Kudo, L. H.; Bell, P. D.; Fowler, B. C.; Corbitt, B. D.; Schafer, J. A.

In: American Journal of Physiology - Renal Fluid and Electrolyte Physiology, Vol. 265, No. 4 34-4, 01.01.1993.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Intracellular Ca 2+ and PKC activation do not inhibit Na + and water transport in rat CCD

AU - Rouch, Alexander

AU - Chen, L.

AU - Kudo, L. H.

AU - Bell, P. D.

AU - Fowler, B. C.

AU - Corbitt, B. D.

AU - Schafer, J. A.

PY - 1993/1/1

Y1 - 1993/1/1

N2 - Experiments examined the effects of elevation of intracellular calcium concentration ([Ca 2+ ](i)) or activation of protein kinase C (PKC) on Na + and water transport in the rat cortical collecting duct (CCD). We measured the lumen-to-bath 22 Na + flux (J(l→b)), transepithelial voltage (V(T)), and water permeability (P(f)) in CCD from deoxycorticosterone (DOC)-treated rats. Ionomycin (0.5 and 1 μM) and thapsigargin (1 and 2 μM) were used to increase [Ca 2+ ](i). Phorbol 12-myristate 13-acetate (PMA; 0.3 and 1 μM) and oleoyl-acetyl-glycerol (OAG; 100 μM) were used as activators of PKC. [Ca 2+ ](i) was measured in isolated perfused tubules using the fluorescent dye fura 2. When added to the bathing solution, 220 pM arginine vasopressin (AVP) failed to affect [Ca 2+ ](i), whereas 1 μM ionomycin increased [Ca 2+ ](i) by 103 ± 15% and 2 μM thapsigargin increased [Ca 2+ ](i) by 24 ± 4%. In flux studies, neither ionomycin nor thapsigargin affected J(l→b) or P(f), although ionomycin caused marked morphological changes. Ionomycin also failed to alter either parameter in tubules from non-DOC-treated rats. Neither 100 μM OAG nor 1 μM PMA affected J(l→b) or P(f). OAG at 50 μM had no effect on V(T) or transepithelial resistance, indicating no inhibition of conductive Na + transport. We conclude that increased [Ca 2+ ](i) and PKC activation do not affect J(l→b) or P(f) in the rat CCD. These findings may account for the sustained increase in J(l→b) produced in the rat CCD by AVP.

AB - Experiments examined the effects of elevation of intracellular calcium concentration ([Ca 2+ ](i)) or activation of protein kinase C (PKC) on Na + and water transport in the rat cortical collecting duct (CCD). We measured the lumen-to-bath 22 Na + flux (J(l→b)), transepithelial voltage (V(T)), and water permeability (P(f)) in CCD from deoxycorticosterone (DOC)-treated rats. Ionomycin (0.5 and 1 μM) and thapsigargin (1 and 2 μM) were used to increase [Ca 2+ ](i). Phorbol 12-myristate 13-acetate (PMA; 0.3 and 1 μM) and oleoyl-acetyl-glycerol (OAG; 100 μM) were used as activators of PKC. [Ca 2+ ](i) was measured in isolated perfused tubules using the fluorescent dye fura 2. When added to the bathing solution, 220 pM arginine vasopressin (AVP) failed to affect [Ca 2+ ](i), whereas 1 μM ionomycin increased [Ca 2+ ](i) by 103 ± 15% and 2 μM thapsigargin increased [Ca 2+ ](i) by 24 ± 4%. In flux studies, neither ionomycin nor thapsigargin affected J(l→b) or P(f), although ionomycin caused marked morphological changes. Ionomycin also failed to alter either parameter in tubules from non-DOC-treated rats. Neither 100 μM OAG nor 1 μM PMA affected J(l→b) or P(f). OAG at 50 μM had no effect on V(T) or transepithelial resistance, indicating no inhibition of conductive Na + transport. We conclude that increased [Ca 2+ ](i) and PKC activation do not affect J(l→b) or P(f) in the rat CCD. These findings may account for the sustained increase in J(l→b) produced in the rat CCD by AVP.

KW - arginine vasopressin

KW - cortical collecting duct

KW - intracellular second messengers

KW - ionomycin

KW - mineralocorticoid

KW - oleoyl-acetyl-glycerol

KW - phorbol 12-myristate 13- acetate

KW - protein kinase C

KW - sodium channel

KW - thapsigargin

KW - vasopressin

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M3 - Article

C2 - 8238386

AN - SCOPUS:0027366737

VL - 265

JO - American Journal of Physiology - Renal Fluid and Electrolyte Physiology

JF - American Journal of Physiology - Renal Fluid and Electrolyte Physiology

SN - 0002-9513

IS - 4 34-4

ER -