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

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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
Creators:
CreatorsEmailORCIDORCID Put Code
Hess, SimonUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Pouzat, ChristopheUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Paeger, LarsUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Pippow, AndreasUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Kloppenburg, PeterUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
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
Uncontrolled Keywords:
KeywordsLanguage
ANTIBIOTIC-STEROL INTERACTIONS; ACHOLEPLASMA-LAIDLAWII CELLS; CALCIUM CURRENT RUNDOWN; RAT SUBSTANTIA-NIGRA; LECITHIN LIPOSOMES; BRAIN-SLICES; GRAMICIDIN-A; WHOLE-CELL; DYNAMICS; DOPAMINEMultiple languages
Cell BiologyMultiple languages
URI: http://kups.ub.uni-koeln.de/id/eprint/58973

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