Enhanced amiloride-sensitive superoxide production in renal medullary thick ascending limb of Dahl salt-sensitive rats

Paul M. O'Connor, Limin Lu, Carlos Schreck, Allen W. Cowley

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

The aims of the present study were to determine whether superoxide (O 2-) production is enhanced in medullary thick ascending limb (mTAL) of Dahl salt-sensitive (SS) rats compared with a salt-resistant consomic control strain (SS.13BN) and to elucidate the cellular pathways responsible for augmented O2- production. Studies were carried out in 7- to 10-wk-old male SS and SS.13BN rats fed either a 0.4% NaCl diet or a 4.0% NaCl diet for 3 days before tissue harvest. Tissue strips containing mTAL were isolated from the left kidney, loaded with the O2--sensitive fluorescent dye dihydroethidium, superfused with modified Hanks' solution, and imaged at X60 magnification on a heated microscope stage. O2- production was stimulated in mTAL by incrementing superfusate NaCl concentration from 154 to 254 to 500 mM. O2- production was enhanced in mTAL of SS rats compared with SS.13BN rats in response to incrementing bath NaCl. Addition of N-methyl-amiloride (100 μM) or inhibition of NAD(P)H oxidase reduced O 2- production in SS mTAL to levels observed in SS.13 BN rats. Both amiloride- and ouabain-sensitive pathways of O 2- production were elevated following 3 days of high (4.0%) NaCl feeding in mTAL of SS and SS.13BN rats. We conclude that mTAL from SS rats exhibit enhanced amiloride-sensitive O2- production. The amiloride-sensitive O2- response in mTAL is independent of active Na+ transport and appears to be mediated by NAD(P)H oxidase. Amiloride-sensitive O2- production is likely to contribute to augmented outer medullary O2- production observed in SS rats during both normal and high NaCl diets.

Original languageEnglish (US)
Pages (from-to)F726-F733
JournalAmerican Journal of Physiology - Renal Physiology
Volume295
Issue number3
DOIs
StatePublished - Sep 1 2008

Fingerprint

Inbred Dahl Rats
Amiloride
Superoxides
Extremities
Inbred BN Rats
Salts
Kidney
NADPH Oxidase
Diet
Active Biological Transport
Ouabain
Baths
Fluorescent Dyes

Keywords

  • Blood pressure
  • Free radicals
  • Hypertension
  • Kidney
  • Reduced nicotinamide adenine dinucleotide phosphatase oxidase

ASJC Scopus subject areas

  • Physiology
  • Urology

Cite this

Enhanced amiloride-sensitive superoxide production in renal medullary thick ascending limb of Dahl salt-sensitive rats. / O'Connor, Paul M.; Lu, Limin; Schreck, Carlos; Cowley, Allen W.

In: American Journal of Physiology - Renal Physiology, Vol. 295, No. 3, 01.09.2008, p. F726-F733.

Research output: Contribution to journalArticle

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abstract = "The aims of the present study were to determine whether superoxide (O 2-) production is enhanced in medullary thick ascending limb (mTAL) of Dahl salt-sensitive (SS) rats compared with a salt-resistant consomic control strain (SS.13BN) and to elucidate the cellular pathways responsible for augmented O2- production. Studies were carried out in 7- to 10-wk-old male SS and SS.13BN rats fed either a 0.4{\%} NaCl diet or a 4.0{\%} NaCl diet for 3 days before tissue harvest. Tissue strips containing mTAL were isolated from the left kidney, loaded with the O2--sensitive fluorescent dye dihydroethidium, superfused with modified Hanks' solution, and imaged at X60 magnification on a heated microscope stage. O2- production was stimulated in mTAL by incrementing superfusate NaCl concentration from 154 to 254 to 500 mM. O2- production was enhanced in mTAL of SS rats compared with SS.13BN rats in response to incrementing bath NaCl. Addition of N-methyl-amiloride (100 μM) or inhibition of NAD(P)H oxidase reduced O 2- production in SS mTAL to levels observed in SS.13 BN rats. Both amiloride- and ouabain-sensitive pathways of O 2- production were elevated following 3 days of high (4.0{\%}) NaCl feeding in mTAL of SS and SS.13BN rats. We conclude that mTAL from SS rats exhibit enhanced amiloride-sensitive O2- production. The amiloride-sensitive O2- response in mTAL is independent of active Na+ transport and appears to be mediated by NAD(P)H oxidase. Amiloride-sensitive O2- production is likely to contribute to augmented outer medullary O2- production observed in SS rats during both normal and high NaCl diets.",
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