PUBLICATION

Improved methods for marking active neuron populations

Authors
Moeyaert, B., Holt, G., Madangopal, R., Perez-Alvarez, A., Fearey, B.C., Trojanowski, N.F., Ledderose, J., Zolnik, T.A., Das, A., Patel, D., Brown, T.A., Sachdev, R.N.S., Eickholt, B.J., Larkum, M.E., Turrigiano, G.G., Dana, H., Gee, C.E., Oertner, T.G., Hope, B.T., Schreiter, E.R.
ID
ZDB-PUB-181027-4
Date
2018
Source
Nature communications   9: 4440 (Journal)
Registered Authors
Ledderose, Julia
Keywords
none
MeSH Terms
  • Animals
  • Antibodies/metabolism
  • Fluorescence
  • HeLa Cells
  • Humans
  • Light
  • Luminescent Proteins/metabolism
  • Mice
  • Neurons/cytology
  • Neurons/metabolism*
  • Protein Engineering/methods*
  • Rats, Wistar
  • Visual Cortex/metabolism
  • Zebrafish/metabolism
PubMed
30361563 Full text @ Nat. Commun.
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.
Genes / Markers
Figures
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Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping