Safiulina 2004 J Neurosci Methods: Difference between revisions
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|authors=Safiulina D, Kaasik A, Seppet E, Peet N, Zharkovsky A, Seppet EK | |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 cellโs 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>ย 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 cellโs 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>ย 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 | ||
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|instruments=Oxygraph-2k | |instruments=Oxygraph-2k | ||
|tissues=Neurons; Brain | |tissues=Neurons; Brain | ||
|preparations=Permeabilized | |preparations=Permeabilized cells | ||
}} | }} |
Revision as of 08:42, 5 April 2012
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 cellโs 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. โข Keywords: Permeabilized neurons, Mitochondrial membrane potential, Mitochondrial calcium, Mitochondrial respiration, Compartmentalization
โข O2k-Network Lab: EE_Tartu_Seppet EK
Labels:
Tissue;cell: Neurons; Brain"Neurons; Brain" is not in the list (Heart, Skeletal muscle, Nervous system, Liver, Kidney, Lung;gill, Islet cell;pancreas;thymus, Endothelial;epithelial;mesothelial cell, Blood cells, Fat, ...) of allowed values for the "Tissue and cell" property. Preparation: Permeabilized cells
HRR: Oxygraph-2k