Dissecting surveying behavior of reactive microglia under chronic neurodegeneration.

In the healthy brain, microglia maintain homeostasis by continuously surveying neuronal health through highly dynamic processes that form purinergic junctions with neuronal somas. These mechanisms are finely tuned for the rapid detection of acute injuries. However, during the transition to a chronically reactive state in neurodegenerative diseases, microglial ramification decreases even as the need for neuronal monitoring escalates. How reactive microglia adapt their surveillance strategies under these conditions remains poorly understood. Using time-lapse imaging of acute brain slices from prion-infected mice, we identified a previously unrecognized mode of neuronal surveillance employed by reactive microglia. Unlike homeostatic microglia, which exhibit low somatic mobility and high process motility, enabling broad, simultaneous monitoring, reactive microglia display high somatic mobility. These cells actively migrate through the brain parenchyma, pausing to form direct and extensive body-to-body contacts with individual neurons. Contact durations ranged from minutes to several hours, often involving partial or full somatic envelopment, with transitions between these states being both frequent and reversible. Notably, reactive microglia exhibited sustained intracellular calcium bursts correlated with their increased mobility. Pharmacological inhibition of the P2Y6 receptor partially reduced microglial migration without disrupting their ability to form neuronal contacts. Furthermore, this highly mobile behavior persisted in acutely isolated reactive microglia