Difference between revisions of "Safiulina 2004 J Neurosci Methods"
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{{Publication | {{Publication | ||
|title=Safiulina D, Kaasik A, Seppet E, Peet N, Zharkovsky A, Seppet | |title=Safiulina D, Kaasik A, Seppet E, Peet N, Zharkovsky A, Seppet EK (2004) Method for in situ detection of the mitochondrial function in neurons. J Neurosci Methods 137:87-95. | ||
|authors=Safiulina D, Kaasik A, Seppet E, Peet N, Zharkovsky A, Seppet | |info=[http://www.ncbi.nlm.nih.gov/pubmed/15196830 PMID: 15196830] | ||
|authors=Safiulina D, Kaasik A, Seppet E, Peet N, Zharkovsky A, Seppet EK | |||
|year=2004 | |year=2004 | ||
|journal=J | |journal=J Neurosci Methods | ||
|abstract=Conventional studies of neuronal mitochondria have been limited to the use of purified preparations of isolated mitochondria, neural cell homogenates, living neurons, or brain slices. However, each technique has several drawbacks. Here, we demonstrate that the neuronal cell’s membrane can be effectively permeabilized by saponin-treatment and that these permeabilized neurons can be used for qualitative and quantitative assessments of oxygen consumption in combination with registration of mitochondrial membrane potential and free Ca<sup>2+</sup> in the matrix. Under these conditions, the mitochondrial function can be studied without removing the mitochondria from their natural milieu thus avoiding the damage of the associated cytoskeleton and outer membrane. At the same time, the method allows the estimation of the mitochondrial function independently of other processes in the cell, and the easy manipulation of the milieu surrounding the mitochondria. Thus, the presented method offers the opportunity to study the neuronal mitochondrial function ''in situ'' and can also be applied to examine the mitochondrial function by other commonly used methods. | |abstract=Conventional studies of neuronal mitochondria have been limited to the use of purified preparations of isolated mitochondria, neural cell homogenates, living neurons, or brain slices. However, each technique has several drawbacks. Here, we demonstrate that the neuronal cell’s membrane can be effectively permeabilized by saponin-treatment and that these permeabilized neurons can be used for qualitative and quantitative assessments of oxygen consumption in combination with registration of mitochondrial membrane potential and free Ca<sup>2+</sup> in the matrix. Under these conditions, the mitochondrial function can be studied without removing the mitochondria from their natural milieu thus avoiding the damage of the associated cytoskeleton and outer membrane. At the same time, the method allows the estimation of the mitochondrial function independently of other processes in the cell, and the easy manipulation of the milieu surrounding the mitochondria. Thus, the presented method offers the opportunity to study the neuronal mitochondrial function ''in situ'' and can also be applied to examine the mitochondrial function by other commonly used methods. | ||
|keywords=Permeabilized neurons, Mitochondrial membrane potential, Mitochondrial calcium, Mitochondrial respiration, Compartmentalization | |keywords=Permeabilized neurons, Mitochondrial membrane potential, Mitochondrial calcium, Mitochondrial respiration, Compartmentalization | ||
| | |mipnetlab=EE Tartu Paju K | ||
}} | }} | ||
{{Labeling | {{Labeling | ||
|tissues= | |tissues=Nervous system | ||
|preparations=Permeabilized | |preparations=Permeabilized cells | ||
|instruments=Oxygraph-2k | |instruments=Oxygraph-2k | ||
}} | }} | ||
Latest revision as of 15:52, 8 June 2015
| Safiulina D, Kaasik A, Seppet E, Peet N, Zharkovsky A, Seppet EK (2004) Method for in situ detection of the mitochondrial function in neurons. J Neurosci Methods 137:87-95. |
Safiulina D, Kaasik A, Seppet E, Peet N, Zharkovsky A, Seppet EK (2004) J Neurosci Methods
Abstract: Conventional studies of neuronal mitochondria have been limited to the use of purified preparations of isolated mitochondria, neural cell homogenates, living neurons, or brain slices. However, each technique has several drawbacks. Here, we demonstrate that the neuronal cell’s membrane can be effectively permeabilized by saponin-treatment and that these permeabilized neurons can be used for qualitative and quantitative assessments of oxygen consumption in combination with registration of mitochondrial membrane potential and free Ca2+ in the matrix. Under these conditions, the mitochondrial function can be studied without removing the mitochondria from their natural milieu thus avoiding the damage of the associated cytoskeleton and outer membrane. At the same time, the method allows the estimation of the mitochondrial function independently of other processes in the cell, and the easy manipulation of the milieu surrounding the mitochondria. Thus, the presented method offers the opportunity to study the neuronal mitochondrial function in situ and can also be applied to examine the mitochondrial function by other commonly used methods. • Keywords: Permeabilized neurons, Mitochondrial membrane potential, Mitochondrial calcium, Mitochondrial respiration, Compartmentalization
• O2k-Network Lab: EE Tartu Paju K
Labels:
Tissue;cell: Nervous system Preparation: Permeabilized cells
HRR: Oxygraph-2k
