Difference between revisions of "Friedrich 2010 Abstract MiP2010"
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{{Abstract | {{Abstract | ||
|title=Kidney cortex mitochondria are non-functional in a potassium-based media whereas heart mitochondria improve with increasing potassium concentration. | |title=Kidney cortex mitochondria are non-functional in a potassium-based media whereas heart mitochondria improve with increasing potassium concentration. | ||
|authors=Malou F, Wilcox CS, Palm F | |authors=Malou F, Wilcox CS, Palm F | ||
|year=2010 | |year=2010 | ||
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Kidney cortex mitochondria in K<sup>+</sup>-based media are non-functional in [K<sup>+</sup>] ranging from 37-146 mM. Heart mitochondria do not display K+-sensitivity to the same degree, but rather increase respiratory function with increasing [K<sup>+</sup>]. Furthermore, we demonstrated that a tissue specific difference in mitochondria K<sup>+</sup>-channels may explain these differences. The present study therefore demonstrates the importance of choosing a correct ''in vitro media'' to ensure a high quality of mitochondria research. | Kidney cortex mitochondria in K<sup>+</sup>-based media are non-functional in [K<sup>+</sup>] ranging from 37-146 mM. Heart mitochondria do not display K+-sensitivity to the same degree, but rather increase respiratory function with increasing [K<sup>+</sup>]. Furthermore, we demonstrated that a tissue specific difference in mitochondria K<sup>+</sup>-channels may explain these differences. The present study therefore demonstrates the importance of choosing a correct ''in vitro media'' to ensure a high quality of mitochondria research. | ||
}} | }} | ||
{{Labeling | {{Labeling | ||
| | |organism=Rat | ||
| | |tissues=Heart, Kidney | ||
| | |model cell lines=Other cell lines | ||
| | |preparations=Isolated mitochondria | ||
|instruments= | |instruments=Oxygraph-2k | ||
|additional=MiP2010 | |additional=MiP2010 | ||
}} | }} | ||
== Affiliation == | == Affiliation == | ||
1-Dept Med Cell Biol, Uppsala Univ, Sweden. | 1-Dept Med Cell Biol, Uppsala Univ, Sweden. | ||
2-Dept Med, Division Hypertension Nephrology, Georgetown Univ Medical Center, Washington DC, USA. - [email protected] | 2-Dept Med, Division Hypertension Nephrology, Georgetown Univ Medical Center, Washington DC, USA. - [email protected] |
Revision as of 09:39, 20 April 2015
Kidney cortex mitochondria are non-functional in a potassium-based media whereas heart mitochondria improve with increasing potassium concentration. |
Link:
Malou F, Wilcox CS, Palm F (2010)
Event: MiP2010
A medium of containing high levels of potassium chloride (KCl) is commonly used when assessing respiratory function of isolated mitochondria from various tissues. However, the measured intracellular [K+] in kidney proximal tubular cells is about 60 mM and in cardiac myocytes approximately 130 mM. Therefore, the use of a similar media [K+] for all tissues seems unsupported. Here we investigated the effect of different [K+] on respiratory function in mitochondria isolated from kidney cortex and heart of healthy male Sprague-Dawley rats.
Oxygen consumptions and the respiratory control ratios (RCR) were measured using respiratory medias containing [K+] of 15, 37, 81, 111 and 146 mM. In all measurements, the media contained (in mM): 1 EGTA, 20 HEPES, 5 MgCl2, 5 KPO4- and 1 g/l bovine serum albumin. pH was adjusted to 7.4 and the osmolarity to 330 mosm/kg HK2O using a 1:3 ratio of sucrose and mannitol.
The RCR of kidney cortex mitochondria decreased when the [K+] was elevated compared to the media containing 15 mM K+ (5.2Β±0.2 vs. 2.5Β±0.2, 3.7Β±0.2, 3.9Β±0.2, 3.0Β±0.1, respectively). However, RCR of heart mitochondria was lowest at 37 mM (3.9Β±0.3) and was highest at 146 mM K+ (10.1Β±0.45). A two-way ANOVA showed that kidney cortex mitochondria have a different sensitivity towards K+ compared to heart mitochondria (interaction P<0.05, treatment P<0.05, group P<0.05). Glibenclamide (100 Β΅M), an inhibitor of the ATP-sensitive K+ channel, increased RCR in kidney cortex mitochondria at 15 mM K+ (+32%), but significantly more at 146 mM K+ (+47%). Blockade of the voltage-gated K+ channel by 4-aminopyridine (4-AP, 1 mM) together with glibenclamide improved RCR by +73% at 146 mM K+. Neither of the applied K+-channel blockers had any effect on the RCR of heart mitochondria. Mitochondria swelling at increasing [K+] were observed in kidney cortex mitochondria, measured as loss of absorbance at 540 nm.
Kidney cortex mitochondria in K+-based media are non-functional in [K+] ranging from 37-146 mM. Heart mitochondria do not display K+-sensitivity to the same degree, but rather increase respiratory function with increasing [K+]. Furthermore, we demonstrated that a tissue specific difference in mitochondria K+-channels may explain these differences. The present study therefore demonstrates the importance of choosing a correct in vitro media to ensure a high quality of mitochondria research.
Labels:
Organism: Rat
Tissue;cell: Heart, Kidney
Preparation: Isolated mitochondria
HRR: Oxygraph-2k
MiP2010
Affiliation
1-Dept Med Cell Biol, Uppsala Univ, Sweden.
2-Dept Med, Division Hypertension Nephrology, Georgetown Univ Medical Center, Washington DC, USA. - [email protected]