Improved methods for marking active neuron populations View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2018-12

AUTHORS

Benjamien Moeyaert, Graham Holt, Rajtarun Madangopal, Alberto Perez-Alvarez, Brenna C. Fearey, Nicholas F. Trojanowski, Julia Ledderose, Timothy A. Zolnik, Aniruddha Das, Davina Patel, Timothy A. Brown, Robert N. S. Sachdev, Britta J. Eickholt, Matthew E. Larkum, Gina G. Turrigiano, Hod Dana, Christine E. Gee, Thomas G. Oertner, Bruce T. Hope, Eric R. Schreiter

ABSTRACT

Marking functionally distinct neuronal ensembles with high spatiotemporal resolution is a key challenge in systems neuroscience. We recently introduced CaMPARI, an engineered fluorescent protein whose green-to-red photoconversion depends on simultaneous light exposure and elevated calcium, which enabled marking active neuronal populations with single-cell and subsecond resolution. However, CaMPARI (CaMPARI1) has several drawbacks, including background photoconversion in low calcium, slow kinetics and reduced fluorescence after chemical fixation. In this work, we develop CaMPARI2, an improved sensor with brighter green and red fluorescence, faster calcium unbinding kinetics and decreased photoconversion in low calcium conditions. We demonstrate the improved performance of CaMPARI2 in mammalian neurons and in vivo in larval zebrafish brain and mouse visual cortex. Additionally, we herein develop an immunohistochemical detection method for specific labeling of the photoconverted red form of CaMPARI. The anti-CaMPARI-red antibody provides strong labeling that is selective for photoconverted CaMPARI in activated neurons in rodent brain tissue. More... »

PAGES

4440

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/s41467-018-06935-2

DOI

http://dx.doi.org/10.1038/s41467-018-06935-2

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1107733469

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/30361563


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Download the RDF metadata as:  json-ld nt turtle xml License info

HOW TO GET THIS DATA PROGRAMMATICALLY:

JSON-LD is a popular format for linked data which is fully compatible with JSON.

curl -H 'Accept: application/ld+json' 'https://scigraph.springernature.com/pub.10.1038/s41467-018-06935-2'

N-Triples is a line-based linked data format ideal for batch operations.

curl -H 'Accept: application/n-triples' 'https://scigraph.springernature.com/pub.10.1038/s41467-018-06935-2'

Turtle is a human-readable linked data format.

curl -H 'Accept: text/turtle' 'https://scigraph.springernature.com/pub.10.1038/s41467-018-06935-2'

RDF/XML is a standard XML format for linked data.

curl -H 'Accept: application/rdf+xml' 'https://scigraph.springernature.com/pub.10.1038/s41467-018-06935-2'


 

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