Fluorescent ratiometric pH indicator SypHer2: applications in neuroscience and regenerative biology

Matlashov, Mikhail E., Bogdanova, Yulia A., Ermakova, Galina V., Mishina, Natalia M., Ermakova, Yulia G., Nikitin, Evgeny S., Balaban, Pavel M., Okabe, Shigeo, Lukyanov, Sergey, Enikolopov, Grigori, Zaraisky, Andrey G., Belousov, Vsevolod V. (August 2015) Fluorescent ratiometric pH indicator SypHer2: applications in neuroscience and regenerative biology. Biochimica et Biophysica Acta (BBA) - General Subjects, 1850 (11). pp. 2318-2328. ISSN 0304-4165

URL: http://www.ncbi.nlm.nih.gov/pubmed/26259819
DOI: 10.1016/j.bbagen.2015.08.002

Abstract

AbstractBackground SypHer is a genetically encoded fluorescent pH-indicator with a ratiometric readout, suitable for measuring fast intracellular pH shifts. However, a relatively low brightness of the indicator limits its use. Methods Here we designed a new version of pH-sensor - SypHer-2, that has up to three times brighter fluorescence signal in cultured mammalian cells compared to the SypHer. Results Using the new indicator we registered activity-associated pH oscillations in neuronal cell culture. We observed prominent temporal neuronal cytoplasm acidification that occurs in parallel with calcium entry. Furthermore, we monitored pH in presynaptic and postsynaptic termini by targeting SypHer-2 directly to these compartments and revealed marked differences in pH dynamics between synaptic boutons and dendritic spines. Finally, we were able to reveal for the first time the intracellular pH drop which occurs within an extended region of the amputated tail of the Xenopus laevis tadpole before it begins to regenerate. Conclusions SypHer2 is suitable for quantitative monitoring of pH in biological systems of different scales, from small cellular subcompartments to animal tissues in vivo. General significance The new pH-sensor will help to investigate pH-dependent processes in both in vitro and in vivo studies.

Item Type: Paper
Subjects: Investigative techniques and equipment
CSHL Authors:
Communities: CSHL labs > Enikopolov lab
Depositing User: Matt Covey
Date: 8 August 2015
Date Deposited: 24 Aug 2015 17:58
Last Modified: 09 Nov 2015 17:03
PMCID: PMC4587288
Related URLs:
URI: http://repository.cshl.edu/id/eprint/31713

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