Analysis of neuronal Ca 2+ handling properties by combining perforated patch clamp recordings and the added buffer approach
Ca functions as an important intracellular signal for a wide range of cellular processes. These processes are selectively activated by controlled spatiotemporal dynamics of the free cytosolic Ca . Intracellular Ca dynamics are regulated by numerous cellular parameters. Here, we established a new way...
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| Veröffentlicht in: | Cell calcium (Edinburgh) 2021-07, Vol.97, p.102411 |
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| container_title | Cell calcium (Edinburgh) |
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| creator | Hess, Simon Pouzat, Christophe Paeger, Lars Pippow, Andreas Kloppenburg, Peter |
| description | Ca
functions as an important intracellular signal for a wide range of cellular processes. These processes are selectively activated by controlled spatiotemporal dynamics of the free cytosolic Ca
. Intracellular Ca
dynamics are regulated by numerous cellular parameters. Here, we established a new way to determine neuronal Ca
handling properties by combining the 'added buffer' approach [1] with perforated patch-clamp recordings [2]. Since the added buffer approach typically employs the standard whole-cell configuration for concentration-controlled Ca
indicator loading, it only allows for the reliable estimation of the immobile fraction of intracellular Ca
buffers. Furthermore, crucial components of intracellular signaling pathways are being washed out during prolonged whole-cell recordings, leading to cellular deterioration. By combining the added buffer approach with perforated patch-clamp recordings, these issues are circumvented, allowing the precise quantification of the cellular Ca
handling properties, including immobile as well as mobile Ca
buffers. |
| doi_str_mv | 10.1016/j.ceca.2021.102411 |
| format | Article |
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functions as an important intracellular signal for a wide range of cellular processes. These processes are selectively activated by controlled spatiotemporal dynamics of the free cytosolic Ca
. Intracellular Ca
dynamics are regulated by numerous cellular parameters. Here, we established a new way to determine neuronal Ca
handling properties by combining the 'added buffer' approach [1] with perforated patch-clamp recordings [2]. Since the added buffer approach typically employs the standard whole-cell configuration for concentration-controlled Ca
indicator loading, it only allows for the reliable estimation of the immobile fraction of intracellular Ca
buffers. Furthermore, crucial components of intracellular signaling pathways are being washed out during prolonged whole-cell recordings, leading to cellular deterioration. By combining the added buffer approach with perforated patch-clamp recordings, these issues are circumvented, allowing the precise quantification of the cellular Ca
handling properties, including immobile as well as mobile Ca
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functions as an important intracellular signal for a wide range of cellular processes. These processes are selectively activated by controlled spatiotemporal dynamics of the free cytosolic Ca
. Intracellular Ca
dynamics are regulated by numerous cellular parameters. Here, we established a new way to determine neuronal Ca
handling properties by combining the 'added buffer' approach [1] with perforated patch-clamp recordings [2]. Since the added buffer approach typically employs the standard whole-cell configuration for concentration-controlled Ca
indicator loading, it only allows for the reliable estimation of the immobile fraction of intracellular Ca
buffers. Furthermore, crucial components of intracellular signaling pathways are being washed out during prolonged whole-cell recordings, leading to cellular deterioration. By combining the added buffer approach with perforated patch-clamp recordings, these issues are circumvented, allowing the precise quantification of the cellular Ca
handling properties, including immobile as well as mobile Ca
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functions as an important intracellular signal for a wide range of cellular processes. These processes are selectively activated by controlled spatiotemporal dynamics of the free cytosolic Ca
. Intracellular Ca
dynamics are regulated by numerous cellular parameters. Here, we established a new way to determine neuronal Ca
handling properties by combining the 'added buffer' approach [1] with perforated patch-clamp recordings [2]. Since the added buffer approach typically employs the standard whole-cell configuration for concentration-controlled Ca
indicator loading, it only allows for the reliable estimation of the immobile fraction of intracellular Ca
buffers. Furthermore, crucial components of intracellular signaling pathways are being washed out during prolonged whole-cell recordings, leading to cellular deterioration. By combining the added buffer approach with perforated patch-clamp recordings, these issues are circumvented, allowing the precise quantification of the cellular Ca
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| ispartof | Cell calcium (Edinburgh), 2021-07, Vol.97, p.102411 |
| issn | 0143-4160 1532-1991 |
| language | eng |
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| source | Elsevier ScienceDirect Journals |
| subjects | Applications Life Sciences Neurobiology Neurons and Cognition Statistics |
| title | Analysis of neuronal Ca 2+ handling properties by combining perforated patch clamp recordings and the added buffer approach |
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