Improved methods for marking active neuron populations
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Improved methods for marking active neuron populations. / Moeyaert, Benjamien; Holt, Graham; Madangopal, Rajtarun; Perez-Alvarez, Alberto; Fearey, Brenna C; Trojanowski, Nicholas F; Ledderose, Julia; Zolnik, Timothy A; Das, Aniruddha; Patel, Davina; Brown, Timothy A; Sachdev, Robert N S; Eickholt, Britta J; Larkum, Matthew E; Turrigiano, Gina G; Dana, Hod; Gee, Christine E; Oertner, Thomas G; Hope, Bruce T; Schreiter, Eric R.
In: NAT COMMUN, Vol. 9, No. 1, 25.10.2018, p. 4440.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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TY - JOUR
T1 - Improved methods for marking active neuron populations
AU - Moeyaert, Benjamien
AU - Holt, Graham
AU - Madangopal, Rajtarun
AU - Perez-Alvarez, Alberto
AU - Fearey, Brenna C
AU - Trojanowski, Nicholas F
AU - Ledderose, Julia
AU - Zolnik, Timothy A
AU - Das, Aniruddha
AU - Patel, Davina
AU - Brown, Timothy A
AU - Sachdev, Robert N S
AU - Eickholt, Britta J
AU - Larkum, Matthew E
AU - Turrigiano, Gina G
AU - Dana, Hod
AU - Gee, Christine E
AU - Oertner, Thomas G
AU - Hope, Bruce T
AU - Schreiter, Eric R
PY - 2018/10/25
Y1 - 2018/10/25
N2 - 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.
AB - 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.
KW - Journal Article
KW - Research Support, Non-U.S. Gov't
U2 - 10.1038/s41467-018-06935-2
DO - 10.1038/s41467-018-06935-2
M3 - SCORING: Journal article
C2 - 30361563
VL - 9
SP - 4440
JO - NAT COMMUN
JF - NAT COMMUN
SN - 2041-1723
IS - 1
ER -