Longitudinal in vivo imaging of perineuronal nets

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 . We introduce a robust approach for quantitative longitudinal imaging of PNNs in brains of awake mice at subcellular resolution. We label PNNs with commercially available compounds and monitor their dynamics with two-photon imaging. Using our approach, we show that it is possible to longitudinally follow the same PNNs while monitoring degradation and reconstitution of PNNs. We demonstrate the compatibility of our method to simultaneously monitor neuronal calcium dynamics and compare the activity of neurons with and without PNNs. Our approach is tailored for studying the intricate role of PNNs , while paving the road for elucidating their role in different neuropathological conditions.