, 2010). Future studies will aim to test the role of complement in microglia-synapse interactions in other CNS regions known to undergo activity-dependent synaptic remodeling. In addition to relevance in global remodeling of circuits

in the healthy brain, our findings have important implications for understanding mechanisms underlying synapse elimination in the diseased brain. Consistent with this idea, abnormal microglia function and complement cascade activation have been associated with neurodegeneration of the CNS (Alexander et al., 2008, Beggs and Salter, 2010, Rosen and Stevens, 2010, Schafer and Stevens, 2010 and Stephan et al., 2012). Indeed, in a mouse model selleck screening library of glaucoma, a neurodegenerative disease associated with RGC loss and gliosis, C1q and C3 are highly upregulated and deposited on retinal synapses and C1q deficiency or microglial “inactivation” with minocycline provide significant neuroprotection (Howell et al., 2011, Steele et al., 2006 and Stevens et al., 2007). In addition to diseases associated with neurodegeneration, recent data from genome-wide association studies and analyses of postmortem human brain tissue have suggested that microglia and/or the complement cascade may also be involved in the development

and pathogenesis of neurodevelopmental and psychiatric disorders (e.g., autism, obsessive compulsive disorder, schizophrenia, etc.) (Chen et al., 2010, Håvik et al., 2011, Monji et al., 2009, Pardo et al., 2005 and Vargas et al., check details 2005). Thus, an intriguing possibility remains that microglia and/or complement dysfunction may be directly involved in diseases associated with synapse loss, dysfunction, and/or development. Together, our data offer insight into mechanisms underlying activity-dependent synaptic pruning in the developing CNS, provide a role for microglia in the healthy brain, and provide important mechanistic insight into microglia-synapse interactions in the healthy and diseased CNS. Selleck Afatinib All experiments were reviewed and overseen by the institutional animal use and care committee in accordance

with all NIH guidelines for the humane treatment of animals. See Supplemental Experimental Procedures for details. Mice, except tdTomato-expressing mice (CHX10-cre::tdTomato), received intraocular injections of anterograde tracers at P4. All mice were sacrificed at P5 and brains were 4% PFA fixed overnight (4°C). Only those brains with sufficient dye fills were analyzed (see Supplemental Experimental Procedures for details). P4 CX3CR1::EGFP heterozygotes were anesthetized with isoflurane and given an intraocular injection of drug (0.5 μM TTX or 10mM forskolin) and vehicle (saline or DMSO) into the left and right eyes, respectively. Injection volume was approximately 200 nl. Four to five hours after first injection, mice received a second intraocular injection of CTB 594 and 647 into the left and right eyes, respectively. Mice were sacrificed at P5 for analysis.