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Fluorogenic Detection of Monoamine Neurotransmitters in Live Cells

Bera, Kallol and Das, Anand Kant and Rakshit, Ananya and Sarkar, Bidyut and Rawat, Anoop and Maity, Barun Kumar and Maiti, Sudipta (2018) Fluorogenic Detection of Monoamine Neurotransmitters in Live Cells. ACS Chemical Neuroscience, 9 (3). pp. 469-474. ISSN 1948-7193. https://resolver.caltech.edu/CaltechAUTHORS:20171211-154134854

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Abstract

Monoamine neurotransmission is key to neuromodulation, but imaging monoamines in live neurons has remained a challenge. Here we show that externally added ortho-phthalaldehyde (OPA) can permeate live cells and form bright fluorogenic adducts with intracellular monoamines (e.g., serotonin, dopamine, and norepinephrine) and with L-DOPA, which can be imaged sensitively using conventional single-photon excitation in a fluorescence microscope. The peak excitation and emission wavelengths (λ_(ex) = 401 nm and λ_(em) = 490 nm for serotonin; λ_(ex) = 446 nm and λ_(em) = 557 nm for dopamine; and λ_(ex) = 446 nm and λ_(em) = 544 nm for norepinephrine, respectively) are accessible to most modern confocal imaging instruments. The identity of monoamine containing structures (possibly neurotransmitter vesicles) in serotonergic RN46A cells is established by quasi-simultaneous imaging of serotonin using three-photon excitation microscopy. Mass spectrometry of cell extracts and of in vitro solutions helps us identify the chemical nature of the adducts and establishes the reaction mechanisms. Our method has low toxicity, high selectivity, and the ability to directly report the location and concentration of monoamines in live cells.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1021/acschemneuro.7b00391DOIArticle
http://pubs.acs.org/doi/abs/10.1021/acschemneuro.7b00391PublisherArticle
https://pubs.acs.org/doi/suppl/10.1021/acschemneuro.7b00391PublisherSupporting Information
ORCID:
AuthorORCID
Rakshit, Ananya0000-0001-8250-0399
Maiti, Sudipta0000-0002-6540-7472
Additional Information:© 2017 American Chemical Society. Received: October 12, 2017; Accepted: December 11, 2017; Published: December 11, 2017. We thank Dr. Bappaditya Chandra (TIFR) and Prof. Soumen Basak (SINP) for their help and support. The authors declare no competing financial interest.
Subject Keywords:serotonin imaging, dopamine imaging, norepinephrine imaging, monoamine microscopy, Falck Hillarp method
Issue or Number:3
Record Number:CaltechAUTHORS:20171211-154134854
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20171211-154134854
Official Citation:Fluorogenic Detection of Monoamine Neurotransmitters in Live Cells. Kallol Bera, Anand Kant Das, Ananya Rakshit, Bidyut Sarkar, Anoop Rawat, Barun Kumar Maity, and Sudipta Maiti. ACS Chemical Neuroscience 2018 9 (3), 469-474 DOI: 10.1021/acschemneuro.7b00391
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:83814
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:12 Dec 2017 21:50
Last Modified:03 Oct 2019 19:11

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