Live imaging of endogenous PSD-95 using ENABLED

A conditional strategy to fluorescently label endogenous proteins

Dale A. Fortin, Shane E. Tillo, Guang Yang, Jong Cheol Rah, Joshua B. Melander, Suxia Bai, Omar Soler-Cedeño, Maozhen Qin, Boris V. Zemelman, Caiying Guo, Tianyi Mao, Haining Zhong

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Stoichiometric labeling of endogenous synaptic proteins for high-contrast live-cell imaging in brain tissue remains challenging. Here, we describe a conditional mouse genetic strategy termed endogenous labeling via exon duplication (ENABLED), which can be used to fluorescently label endogenous proteins with near ideal properties in all neurons, a sparse subset of neurons, or specific neuronal subtypes. We used this method to label the postsynaptic density protein PSD-95 with mVenus without overexpression side effects. We demonstrated that mVenus-tagged PSD-95 is functionally equivalent to wild-type PSD-95 and that PSD-95 is present in nearly all dendritic spines in CA1 neurons. Within spines, while PSD-95 exhibited low mobility under basal conditions, its levels could be regulated by chronic changes in neuronal activity. Notably, labeled PSD-95 also allowed us to visualize and unambiguously examine otherwiseunidentifiable excitatory shaft synapses in aspiny neurons, such as parvalbumin-positive interneurons and dopaminergic neurons. Our results demonstrate that the ENABLED strategy provides a valuable new approach to study the dynamics of endogenous synaptic proteins in vivo.

Original languageEnglish (US)
Pages (from-to)16698-16712
Number of pages15
JournalJournal of Neuroscience
Volume34
Issue number50
DOIs
StatePublished - Dec 10 2014

Fingerprint

Exons
Neurons
Proteins
Parvalbumins
Dendritic Spines
Dopaminergic Neurons
Interneurons
Neuroimaging
Synapses
Spine

Keywords

  • Cell-type-specific labeling
  • Conditional knock-in
  • Live imaging
  • Protein labeling
  • Psd-95
  • Sparse labeling

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Live imaging of endogenous PSD-95 using ENABLED : A conditional strategy to fluorescently label endogenous proteins. / Fortin, Dale A.; Tillo, Shane E.; Yang, Guang; Rah, Jong Cheol; Melander, Joshua B.; Bai, Suxia; Soler-Cedeño, Omar; Qin, Maozhen; Zemelman, Boris V.; Guo, Caiying; Mao, Tianyi; Zhong, Haining.

In: Journal of Neuroscience, Vol. 34, No. 50, 10.12.2014, p. 16698-16712.

Research output: Contribution to journalArticle

Fortin, DA, Tillo, SE, Yang, G, Rah, JC, Melander, JB, Bai, S, Soler-Cedeño, O, Qin, M, Zemelman, BV, Guo, C, Mao, T & Zhong, H 2014, 'Live imaging of endogenous PSD-95 using ENABLED: A conditional strategy to fluorescently label endogenous proteins', Journal of Neuroscience, vol. 34, no. 50, pp. 16698-16712. https://doi.org/10.1523/JNEUROSCI.3888-14.2014
Fortin, Dale A. ; Tillo, Shane E. ; Yang, Guang ; Rah, Jong Cheol ; Melander, Joshua B. ; Bai, Suxia ; Soler-Cedeño, Omar ; Qin, Maozhen ; Zemelman, Boris V. ; Guo, Caiying ; Mao, Tianyi ; Zhong, Haining. / Live imaging of endogenous PSD-95 using ENABLED : A conditional strategy to fluorescently label endogenous proteins. In: Journal of Neuroscience. 2014 ; Vol. 34, No. 50. pp. 16698-16712.
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