Main Menu (Mobile)- Block

Main Menu - Block

Improved methods for marking active neuron populations.

Our Research / Publications
janelia7_blocks-janelia7_biblio_header | block
Nature Communications. 2018 Oct 25;9(1):4440. doi: 10.1038/s41467-018-06935-2
View Article on
Improved methods for marking active neuron populations. Schreiter LabGENIETool Translation Team (T3)
Moeyaert B, Holt G, Madangopal R, Perez-Alvarez A, Fearey BC, Trojanowski NF, Ledderose J, Zolnik TA, Das A, Patel D, Brown TA, Sachdev RN, Eickholt BJ, Larkum ME, Turrigiano GG, Dana H, Gee CE, Oertner TG, Hope BT, Schreiter ER
janelia7_blocks-janelia7_biblio_abstract | block

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.

PMID: 30361563 [PubMed - indexed for MEDLINE]

node:body | entity_field
janelia7_blocks-janelia7_biblio_authors | block

Janelia Authors

Tim Brown
Eric Schreiter
janelia7_blocks-janelia7_biblio_tools | block