Cited by 46Mitochondrial Function and PathologyIon channel regulation and functionATP Synthase and ATPases Research

Lack of manifestations of diazoxide/5‐hydroxydecanoate‐sensitive KATP channel in rat brain nonsynaptosomal mitochondria

Brustovetsky, Tatiana, Shalbuyeva, Natalia, Brustovetsky, Nickolay·2005·The Journal of Physiology

Abstract

Pharmacological modulation of the mitochondrial ATP‐sensitive K+ channel (mitoK ATP) sensitive to diazoxide and 5‐hydroxydecanoate (5‐HD) represents an attractive strategy to protect cells against ischaemia/reperfusion‐ and stroke‐related injury. To re‐evaluate a functional role for the mitoK ATP in brain, we used Percoll‐gradient‐purified brain nonsynaptosomal mitochondria in a light absorbance assay, in radioisotope measurements of matrix volume, and in measurements of respiration, membrane potential (ΔΨ) and depolarization‐induced K+ efflux. The changes in mitochondrial morphology were evaluated by transmission electron microscopy (TEM). Polyclonal antibodies raised against certain fragments of known sulphonylurea receptor subunits, SUR1 and SUR2, and against different epitopes of K+ inward rectifier subunits Kir 6.1 and Kir 6.2 of the ATP‐sensitive K+ channel of the plasma membrane (cellK ATP), were employed to detect similar subunits in brain mitochondria. A variety of plausible blockers (ATP, 5‐hydroxydecanoate, glibenclamide, tetraphenylphosphonium cation) and openers (diazoxide, pinacidil, chromakalim, minoxidil, testosterone) of the putative mitoK ATP were applied to show the role of the channel in regulating matrix volume, respiration, and ΔΨ and K+ fluxes across the inner mitochondrial membrane. None of the pharmacological agents applied to brain mitochondria in the various assays pinpointed processes that could be unequivocally associated with mitoK ATP activity. In addition, immunoblotting analysis did not provide explicit evidence for the presence of the mitoK ATP, similar to the cellK ATP, in brain mitochondria. On the other hand, the depolarization‐evoked release of K+ suppressed by ATP could be re‐activated by carboxyatractyloside, an inhibitor of the adenine nucleotide translocase (ANT). Moreover, bongkrekic acid, another inhibitor of the ANT, inhibited K+ efflux similarly to ATP. These observations implicate the ANT in ATP‐sensitive K+ transport in brain mitochondria.

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Crossref · 10.1113/jphysiol.2005.091199

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Authors
Brustovetsky, T., Shalbuyeva, N., & Brustovetsky, N.
Affiliations
  • Indiana University School of Medicine (US)
  • Indiana University Indianapolis (US)
Venue
The Journal of Physiology
Publisher
Wiley
Published
2005-09-28
Volume
568
Issue
1
Pages
47-59
ISSN
  • 0022-3751 (ISSN-L)
  • 1469-7793
OpenAlex
W1597339748
Cited by
46
References
59
Citations by year
2022: 1 · 2021: 1 · 2019: 2 · 2017: 1 · 2016: 1 · 2015: 2 · 2014: 1 · 2013: 4 · 2012: 5
FWCI
2.64
Type
journal article
Language
en
Open access
closed
Locations
  • The Journal of Physiology (publishedVersion)
  • PubMed (publishedVersion)
Funders
National Institute of Neurological Disorders and Stroke (R01NS050131)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Mitochondrial Function and Pathology
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ion channel regulation and function
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › ATP Synthase and ATPases Research
Keywords
mitochondrial ATP-sensitive potassium channel, diazoxide, 5-hydroxydecanoate, brain mitochondria, sulfonylurea receptor, Kir6.1, Kir6.2, mitochondrial membrane potential, adenine nucleotide translocase, carboxyatractyloside, bongkrekic acid, ATP, mitochondrial respiration, ischemia-reperfusion injury, stroke
MeSH terms
Kcnj11 Channel, Mitochondrial ADP, ATP Translocases, Adenosine Triphosphate, Animals, Atractyloside, Brain, Culture Media, Decanoic Acids, Diazoxide, Hydroxy Acids, Male, Mitochondria, Oxygen Consumption, Potassium, Potassium Chloride, Receptors, Drug, Potassium Channels, Rats, Sprague-Dawley, ATP-Binding Cassette Transporters, Potassium Channels, Inwardly Rectifying, Potassium Channel Blockers, Rats, KATP Channels, Sulfonylurea Receptors, In Vitro Techniques

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