Deep coverage microscopy exposes a pharmacological window for modifiers of neuronal network connectivity

Abstract

BackgroundTherapeutic developments for neurodegenerative disorders are redirecting their focus to the mechanisms that contribute to synaptic plasticity and the loss thereof. Identification of novel regulators requires a method to quantify neuronal network connectivity with high accuracy and throughput. To meet this demand, we have established a microscopy-based pipeline that integrates morphological and functional correlates of connectivity in primary neuronal culture. ResultsWe unveiled a connectivity signature that was specific to the cell type and culture age. We defined a score that accurately reports on the degree of neuronal connectivity and we validated this score by targeted perturbation of microtubule stability and selective depletion of anti-oxidants. With a focused compound screen, we discovered that inhibition of dual leucine zipper kinase activity increased neuronal connectivity in otherwise unperturbed cultures and exerted neuroprotective effects in cultures grown under sub-optimal or challenged conditions. ConclusionsOur results illustrate that profiling microscopy images with deep coverage enables sensitive interrogation of neuronal connectivity and allows exposing a dose and time window for pharmacological interventions. Therefore, the current approach holds promise for identifying pathways and compounds that preserve or rescue neuronal connectivity in neurodegenerative disorders.