Longitudinal in vivo imaging of perineuronal nets

Amit Benbenishty, Shany Peled-Hajaj, Venkat Raghavan Krishnaswamy, Hagai Har-Gil, Sapir Havusha-Laufer, Antonella Ruggiero, Inna Slutsky, Pablo Blinder*, Irit Sagi*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

1 Scopus citations


Significance: Perineuronal nets (PNNs) are extracellular matrix structures implicated in learning, memory, information processing, synaptic plasticity, and neuroprotection. However, our understanding of mechanisms governing the evidently important contribution of PNNs to central nervous system function is lacking. A primary cause for this gap of knowledge is the absence of direct experimental tools to study their role in vivo. Aim: We introduce a robust approach for quantitative longitudinal imaging of PNNs in brains of awake mice at subcellular resolution. Approach: We label PNNs in vivo with commercially available compounds and monitor their dynamics with two-photon imaging. Results: Using our approach, we show that it is possible to longitudinally follow the same PNNs in vivo while monitoring degradation and reconstitution of PNNs. We demonstrate the compatibility of our method to simultaneously monitor neuronal calcium dynamics in vivo and compare the activity of neurons with and without PNNs. Conclusion: Our approach is tailored for studying the intricate role of PNNs in vivo, while paving the road for elucidating their role in different neuropathological conditions.

Original languageEnglish
Article number015008
Issue number1
StatePublished - 1 Jan 2023


FundersFunder number
European Research Council639416, DLV-695437
United States-Israel Binational Science Foundation712506-01
Israel Science Foundation1226/13, 1994/15, 1019/15
Azrieli Foundation


    • extracellular matrix
    • intravital
    • parvalbumin
    • perineuronal nets
    • two-photon imaging


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