Simultaneous two-photon imaging of intracellular chloride concentration and pH in mouse pyramidal neurons in vivo

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Sato , S S , Artoni , P , Landi , S , Cozzolino , O , Parra , R , Pracucci , E , Trovato , F , Szczurkowska , J , Luin , S , Arosio , D , Beltram , F , Cancedda , L , Kaila , K & Ratto , G M 2017 , ' Simultaneous two-photon imaging of intracellular chloride concentration and pH in mouse pyramidal neurons in vivo ' , Proceedings of the National Academy of Sciences of the United States of America , vol. 114 , no. 41 , pp. E8770-E8779 . https://doi.org/10.1073/pnas.1702861114

Title: Simultaneous two-photon imaging of intracellular chloride concentration and pH in mouse pyramidal neurons in vivo
Author: Sato, Sebastian Sulis; Artoni, Pietro; Landi, Silvia; Cozzolino, Olga; Parra, Riccardo; Pracucci, Enrico; Trovato, Francesco; Szczurkowska, Joanna; Luin, Stefano; Arosio, Daniele; Beltram, Fabio; Cancedda, Laura; Kaila, Kai; Ratto, Gian Michele
Contributor organization: Biosciences
Kai Kaila / Principal Investigator
Neuroscience Center
Physiology and Neuroscience (-2020)
Helsinki Institute of Life Science HiLIFE, Common
Laboratory of Neurobiology
Date: 2017-10-10
Language: eng
Number of pages: 10
Belongs to series: Proceedings of the National Academy of Sciences of the United States of America
ISSN: 0027-8424
DOI: https://doi.org/10.1073/pnas.1702861114
URI: http://hdl.handle.net/10138/305660
Abstract: Intracellular chloride ([Cl-](i)) andpH(pH(i)) are fundamental regulators of neuronal excitability. They exert wide-ranging effects on synaptic signaling and plasticity and on development and disorders of the brain. The ideal technique to elucidate the underlying ionic mechanisms is quantitative and combined two-photon imaging of [Cl-](i) and pH(i), but this has never been performed at the cellular level in vivo. Here, by using a genetically encoded fluorescent sensor that includes a spectroscopic reference (an element insensitive to Cl-and pH), we show that ratiometric imaging is strongly affected by the optical properties of the brain. We have designed a method that fully corrects for this source of error. Parallel measurements of [Cl-](i) and pH(i) at the single-cell level in the mouse cortex showed the in vivo presence of the widely discussed developmental fall in [Cl-](i) and the role of the K-Cl cotransporter KCC2 in this process. Then, we introduce a dynamic two-photon excitation protocol to simultaneously determine the changes of pHi and [Cl-](i) in response to hypercapnia and seizure activity.
Subject: fluorescent indicators
neurodevelopment
neuronal inhibition
bumetanide
NKCC1
SYNAPTIC INHIBITION
HIPPOCAMPAL-NEURONS
COTRANSPORTER KCC2
CA1 NEURONS
GFP
BRAIN
EXCITATION
PLASTICITY
INDICATOR
EPILEPSY
3112 Neurosciences
Peer reviewed: Yes
Rights: other
Usage restriction: openAccess
Self-archived version: publishedVersion


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