Intracellular diffusion restrictions in isolated cardiomyocytes from rainbow trout

<p>Abstract</p> <p>Background</p> <p>Restriction of intracellular diffusion of adenine nucleotides has been studied intensively on adult rat cardiomyocytes. However, their cause and role <it>in vivo </it>is still uncertain. Intracellular membrane structures...

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Main Authors: Birkedal Rikke, Vendelin Marko, Sokolova Niina
Format: Article
Language:English
Published: BMC 2009-12-01
Series:BMC Cell Biology
Online Access:http://www.biomedcentral.com/1471-2121/10/90
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author Birkedal Rikke
Vendelin Marko
Sokolova Niina
author_facet Birkedal Rikke
Vendelin Marko
Sokolova Niina
author_sort Birkedal Rikke
collection DOAJ
description <p>Abstract</p> <p>Background</p> <p>Restriction of intracellular diffusion of adenine nucleotides has been studied intensively on adult rat cardiomyocytes. However, their cause and role <it>in vivo </it>is still uncertain. Intracellular membrane structures have been suggested to play a role. We therefore chose to study cardiomyocytes from rainbow trout (<it>Oncorhynchus mykiss</it>), which are thinner and have fewer intracellular membrane structures than adult rat cardiomyocytes. Previous studies suggest that trout permeabilized cardiac fibers also have diffusion restrictions. However, results from fibers may be affected by incomplete separation of the cells. This is avoided when studying permeabilized, isolated cardiomyocytes. The aim of this study was to verify the existence of diffusion restrictions in trout cardiomyocytes by comparing ADP-kinetics of mitochondrial respiration in permeabilized fibers, permeabilized cardiomyocytes and isolated mitochondria from rainbow trout heart. Experiments were performed at 10, 15 and 20°C in the absence and presence of creatine.</p> <p>Results</p> <p>Trout cardiomyocytes hypercontracted in the solutions used for mammalian cardiomyocytes. We developed a new solution in which they retained their shape and showed stable steady state respiration rates throughout an experiment. The apparent ADP-affinity of permeabilized cardiomyocytes was different from that of fibers. It was higher, independent of temperature and not increased by creatine. However, it was still about ten times lower than in isolated mitochondria.</p> <p>Conclusions</p> <p>The differences between fibers and cardiomyocytes suggest that results from trout heart fibers were affected by incomplete separation of the cells. However, the lower ADP-affinity of cardiomyocytes compared to isolated mitochondria indicate that intracellular diffusion restrictions are still present in trout cardiomyocytes despite their lower density of intracellular membrane structures. The lack of a creatine effect indicates that trout heart lacks mitochondrial creatine kinase tightly coupled to respiration. This argues against diffusion restriction by the outer mitochondrial membrane. These results from rainbow trout cardiomyocytes resemble those from other low-performance hearts such as neonatal rat and rabbit hearts. Thus, it seems that metabolic regulation is related to cardiac performance, and it is likely that rainbow trout can be used as a model animal for further studies of the localization and role of diffusion restrictions in low-performance hearts.</p>
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spelling doaj.art-0a2c3937e072473c87476df0ecfb8e9b2022-12-22T02:50:11ZengBMCBMC Cell Biology1471-21212009-12-011019010.1186/1471-2121-10-90Intracellular diffusion restrictions in isolated cardiomyocytes from rainbow troutBirkedal RikkeVendelin MarkoSokolova Niina<p>Abstract</p> <p>Background</p> <p>Restriction of intracellular diffusion of adenine nucleotides has been studied intensively on adult rat cardiomyocytes. However, their cause and role <it>in vivo </it>is still uncertain. Intracellular membrane structures have been suggested to play a role. We therefore chose to study cardiomyocytes from rainbow trout (<it>Oncorhynchus mykiss</it>), which are thinner and have fewer intracellular membrane structures than adult rat cardiomyocytes. Previous studies suggest that trout permeabilized cardiac fibers also have diffusion restrictions. However, results from fibers may be affected by incomplete separation of the cells. This is avoided when studying permeabilized, isolated cardiomyocytes. The aim of this study was to verify the existence of diffusion restrictions in trout cardiomyocytes by comparing ADP-kinetics of mitochondrial respiration in permeabilized fibers, permeabilized cardiomyocytes and isolated mitochondria from rainbow trout heart. Experiments were performed at 10, 15 and 20°C in the absence and presence of creatine.</p> <p>Results</p> <p>Trout cardiomyocytes hypercontracted in the solutions used for mammalian cardiomyocytes. We developed a new solution in which they retained their shape and showed stable steady state respiration rates throughout an experiment. The apparent ADP-affinity of permeabilized cardiomyocytes was different from that of fibers. It was higher, independent of temperature and not increased by creatine. However, it was still about ten times lower than in isolated mitochondria.</p> <p>Conclusions</p> <p>The differences between fibers and cardiomyocytes suggest that results from trout heart fibers were affected by incomplete separation of the cells. However, the lower ADP-affinity of cardiomyocytes compared to isolated mitochondria indicate that intracellular diffusion restrictions are still present in trout cardiomyocytes despite their lower density of intracellular membrane structures. The lack of a creatine effect indicates that trout heart lacks mitochondrial creatine kinase tightly coupled to respiration. This argues against diffusion restriction by the outer mitochondrial membrane. These results from rainbow trout cardiomyocytes resemble those from other low-performance hearts such as neonatal rat and rabbit hearts. Thus, it seems that metabolic regulation is related to cardiac performance, and it is likely that rainbow trout can be used as a model animal for further studies of the localization and role of diffusion restrictions in low-performance hearts.</p>http://www.biomedcentral.com/1471-2121/10/90
spellingShingle Birkedal Rikke
Vendelin Marko
Sokolova Niina
Intracellular diffusion restrictions in isolated cardiomyocytes from rainbow trout
BMC Cell Biology
title Intracellular diffusion restrictions in isolated cardiomyocytes from rainbow trout
title_full Intracellular diffusion restrictions in isolated cardiomyocytes from rainbow trout
title_fullStr Intracellular diffusion restrictions in isolated cardiomyocytes from rainbow trout
title_full_unstemmed Intracellular diffusion restrictions in isolated cardiomyocytes from rainbow trout
title_short Intracellular diffusion restrictions in isolated cardiomyocytes from rainbow trout
title_sort intracellular diffusion restrictions in isolated cardiomyocytes from rainbow trout
url http://www.biomedcentral.com/1471-2121/10/90
work_keys_str_mv AT birkedalrikke intracellulardiffusionrestrictionsinisolatedcardiomyocytesfromrainbowtrout
AT vendelinmarko intracellulardiffusionrestrictionsinisolatedcardiomyocytesfromrainbowtrout
AT sokolovaniina intracellulardiffusionrestrictionsinisolatedcardiomyocytesfromrainbowtrout