Hess, Simon, Pouzat, Christophe, Paeger, Lars, Pippow, Andreas and Kloppenburg, Peter (2021). Analysis of neuronal Ca2+ handling properties by combining perforated patch clamp recordings and the added buffer approach. Cell Calcium, 97. OXFORD: ELSEVIER SCI LTD. ISSN 1532-1991
Full text not available from this repository.Abstract
Ca2+ 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 Ca2+. Intracellular Ca2+ dynamics are regulated by numerous cellular parameters. Here, we established a new way to determine neuronal Ca2+ 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 Ca2+ indicator loading, it only allows for the reliable estimation of the immobile fraction of intracellular Ca2+ 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 Ca2+ handling properties, including immobile as well as mobile Ca2+ buffers.
Item Type: | Journal Article | ||||||||||||||||||||||||
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URN: | urn:nbn:de:hbz:38-589730 | ||||||||||||||||||||||||
DOI: | 10.1016/j.ceca.2021.102411 | ||||||||||||||||||||||||
Journal or Publication Title: | Cell Calcium | ||||||||||||||||||||||||
Volume: | 97 | ||||||||||||||||||||||||
Date: | 2021 | ||||||||||||||||||||||||
Publisher: | ELSEVIER SCI LTD | ||||||||||||||||||||||||
Place of Publication: | OXFORD | ||||||||||||||||||||||||
ISSN: | 1532-1991 | ||||||||||||||||||||||||
Language: | English | ||||||||||||||||||||||||
Faculty: | Unspecified | ||||||||||||||||||||||||
Divisions: | Unspecified | ||||||||||||||||||||||||
Subjects: | no entry | ||||||||||||||||||||||||
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URI: | http://kups.ub.uni-koeln.de/id/eprint/58973 |
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