Oxidative stress increases SNAT1 expression and stimulates cysteine uptake in freshly isolated rat cardiomyocytes

N King, H Lin, M-S Suleiman

Research output: Contribution to journalArticle (Academic Journal)peer-review

20 Citations (Scopus)

Abstract

Intracellular cysteine availability is an important rate-limiting factor governing glutathione synthesis in the heart. This is also dependent on the magnitude and rate of cysteine uptake into cardiomyocytes, which has been little studied. This study investigated the hypothesis that changes to cysteine transporter expression and activity during oxidative stress influence cardiomyocyte glutathione levels. The uptake of 0-3 mM L: -[(35)S]cysteine into ventricular cardiomyocytes isolated from adult male Wistar rats was measured using oil filtration. Cysteine transporter expression was investigated by conventional and real-time quantitative reverse-transcription polymerase chain reaction and Western blotting. Glutathione levels were measured enzymatically. Oxidative stress was induced via 0-6 h incubation with 0.05 mM H(2)O(2). Cysteine uptake was greatest in sodium-containing media and was inhibited by glutamine, 2-(methylamino)-isobutyric acid (alphaMeAIB), serine or alanine. The K (m) and V (max) of the alphaMeAIB insensitive and sensitive portions were 0.133 +/- 0.01 mM and 468.11 +/- 9.04 pmol/mul cell vol/min, and 0.557 +/- 0.096 mM and 279.87 +/- 16.06 pmol/mul cell vol/min, respectively. Cardiomyocytes expressed ASCT2, SNAT1 and SNAT2 but not ASCT1. Oxidative stress significantly enhanced cysteine uptake, which was attenuated by alphaMeAIB. This was accompanied by significantly enhanced SNAT1 expression, whilst SNAT2 and ASCT2 were unaffected. Incubation with cysteine significantly reduced the oxidative-stress-induced decline in cardiomyocyte glutathione as compared to cells incubated without cysteine or cells incubated with cysteine and alphaMeAIB. In conclusion, under control conditions SNAT transporters aid in the delivery of cysteine for cardiomyocyte GSH synthesis, whilst oxidative stress increases cardiomyocyte cysteine uptake and stimulates cardiomyocyte SNAT1 expression.
Translated title of the contributionOxidative stress increases SNAT1 expression and stimulates cysteine uptake in freshly isolated rat cardiomyocytes
Original languageEnglish
Pages (from-to)517 - 526
Number of pages10
JournalAmino Acids
Volume40
Issue number2
DOIs
Publication statusPublished - Feb 2011

Bibliographical note

Publisher: SpringerLink

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