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Microglial P2Y12 mediates chronic stress-induced synapse loss in the prefrontal cortex and associated behavioral consequences


Chronic unpredictable stress (CUS) drives microglia-mediated neuronal remodeling and synapse loss in the prefrontal cortex (PFC), contributing to deficits in cognition and behavior. However, it remains unclear what mechanisms guide microglia-neuron interactions in stress. Evidence indicates that neuronal activity-dependent purinergic signaling directs microglial processes and synaptic engagement via P2Y12, a purinergic receptor exclusively expressed by microglia in the brain. Stress alters excitatory neurotransmission in the PFC, thus we aimed to determine if P2Y12 signaling promotes functional changes in microglia in chronic stress. Here we used genetic ablation of P2Y12 (P2ry12–/–) or pharmacological blockade (clopidogrel, ticagrelor) to examine the role of purinergic signaling in stress-induced microglia-neuron interaction. Multiple behavioral, physiological, and cytometric endpoints were analyzed. Deletion of P2Y12 led to a number of fundamental alterations in the PFC, including the heightened microglial number and increased dendritic spine density. Flow cytometry revealed that microglia in P2ry12–/– mice had shifts in surface levels of CX3CR1, CSF1R, and CD11b, suggesting changes in synaptic engagement and phagocytosis in the PFC. In line with this, pharmacological blockade of P2Y12 prevented CUS-induced increases in the proportion of microglia with neuronal inclusions, limited dendritic spine loss in the PFC, and attenuated alterations in stress coping behavior and working memory function. Overall, these findings indicate that microglial P2Y12 is a critical mediator of stress-induced synapse loss in the PFC and subsequent behavioral deficits.

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Fig. 1: Constitutive ablation of P2Y12 attenuates stress effects on coping behavior and working memory, and induces significant alterations in frontal cortex microglia.
Fig. 2: Loss of P2Y12 increases lysosome markers in the medial prefrontal cortex and prevents dendritic remodeling in chronic stress.
Fig. 3: Pharmacological blockade of microglial P2Y12 attenuates stress effects on behavior and shifts microglial phenotype in the frontal cortex.
Fig. 4: Administration of clopidogrel reduces levels of microglial P2Y12 and prevents stress effects on microglial morphology.
Fig. 5: Antagonism of microglial P2Y12 prevents stress-induced phagocytosis of dendritic elements and subsequent dendritic spine loss in the mPFC.


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The authors would like to thank Dr. Ania Majewska for kindly donating P2ry12–/– mice, and Dr. Grayson Sipe for advice regarding administration of clopidogrel and ticagrelor. The authors would also like to thank Dr. Lauren Vollmer for feedback on this manuscript. Additional support was provided by the Statistics Consulting Center at the University of Cincinnati. This work was supported by the National Institute of Mental Health (F32MH123051, JLB; R01MH123545, ESW) and the University of Cincinnati.

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Study conception and design: JLB, ESW. Acquisition of data: JLB, DTD, JKF, ILR, SCW. Analysis and interpretation of data: JLB, ESW. Drafting of paper: JLB, ESW.

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Correspondence to Eric S. Wohleb.

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Bollinger, J.L., Dadosky, D.T., Flurer, J.K. et al. Microglial P2Y12 mediates chronic stress-induced synapse loss in the prefrontal cortex and associated behavioral consequences. Neuropsychopharmacol. (2022).

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