The formation of mature neural circuits requires selective pruning of inappropriate synapses and strengthening of appropriate synaptic connections. A longstanding question in neurobiology is what determines which synapses will be eliminated? While the role of spontaneous and experience-driven synaptic activity in developmental synaptic pruning is well established, surprisingly little is known about the molecules and mechanisms that link neural activity with the physical elimination of specific synapses. Our recent studies reveal that glial cells-microglia and astrocytes--are key players in developmental synaptic pruning. We discovered that C1q, the initiating protein of the classical complement cascade, is significantly upregulated in developing neurons by immature astrocytes. Both C1q and downstream C3 are moreover localized to synapses and are required for developmental synapse elimination in the visual system;however the mechanisms by which complement mediates synaptic pruning are completely unknown. The primary role of the complement cascade in the innate immune system is to opsonize or tag unwanted cells or debris for removal by phagocytic macrophages via specific complement receptors. Our preliminary studies support a model in which inappropriate synapses in the developing brain are similarly tagged by complement and then eliminated by microglia, the primary phagocytic cells in CNS. Given the importance of neural activity in synaptic pruning, a major goal of the proposed research is to determine whether and how the complement system cooperates with neuronal activity to give rise to precise visual circuit wiring. Activity-dependent competition between neighboring axons is thought to drive the elimination of weak synaptic inputs;however, the molecular mechanisms remain elusive. We propose a model in which activity and astrocyte-derived factor(s) act cooperatively to upregulate C1q in developing neurons which leads to local activation of the complement cascade at neighboring weak synapses and the elimination of complement (C3) tagged synapses by microglia. We will use the mouse retinogeniculate system as a model to manipulate neural activity at specific synapses in vivo to examine the role of microglia (Aim 1), C1q and C3 (Aim 2) and astrocytes (Aim 3) in activity-dependent synapse elimination. Specifically, we will ask: 1) Does complement specifically tag weak synapses for elimination? 2) Do microglia actively prune synapses or engulf synapses already undergoing elimination? 3) Does neuronal activity regulate the complement cascade, and if so, how? The answers to these questions will add to our understanding of the role of microglia and the complement cascade in developmental synapse elimination, and may ultimately provide new insight into how CNS synapses are eliminated during normal brain wiring, and possibly in diseases involving aberrant synapse loss and synaptic connectivity, such as epilepsy and autism.
The formation of mature neural circuits requires selective pruning of inappropriate synapses and strengthening of appropriate synaptic connections. Disruption of neural activity during the postnatal development leads to permanent defects in synaptic connectivity;however, a major gap in our knowledge is the identity of the molecules and mechanisms that link neural activity with the physical elimination of specific synapses. Defects in synapse elimination cause imbalances in excitation and inhibition in the nervous system, possibly leading to neurological disorders such as autism and epilepsy. In addition, synapse loss and dysfunction are becoming increasingly recognized as a hallmark of many CNS neurodegenerative diseases. Our preliminary studies reveal that glial cells-microglia and astrocytes--are key players in activity-dependent synapse elimination A thorough knowledge of the cellular and molecular mechanisms by which glial cells control normal developmental synaptic pruning could provide mechanistic insight into how to prevent aberrant synapse loss during CNS neurodevelopmental disorders and disease.
|Hong, Soyon; Wilton, Daniel K; Stevens, Beth et al. (2017) Structured Illumination Microscopy for the Investigation of Synaptic Structure and Function. Methods Mol Biol 1538:155-167|
|Loh, Ken H; Stawski, Philipp S; Draycott, Austin S et al. (2016) Proteomic Analysis of Unbounded Cellular Compartments: Synaptic Clefts. Cell 166:1295-1307.e21|
|Schafer, Dorothy P; Heller, Christopher T; Gunner, Georgia et al. (2016) Microglia contribute to circuit defects in Mecp2 null mice independent of microglia-specific loss of Mecp2 expression. Elife 5:|
|Chung, Won-Suk; Welsh, Christina A; Barres, Ben A et al. (2015) Do glia drive synaptic and cognitive impairment in disease? Nat Neurosci 18:1539-1545|
|Schafer, Dorothy P; Stevens, Beth (2015) Microglia Function in Central Nervous System Development and Plasticity. Cold Spring Harb Perspect Biol 7:a020545|
|Wu, Yuwen; Dissing-Olesen, Lasse; MacVicar, Brian A et al. (2015) Microglia: Dynamic Mediators of Synapse Development and Plasticity. Trends Immunol 36:605-13|
|Bilimoria, Parizad M; Stevens, Beth (2015) Microglia function during brain development: New insights from animal models. Brain Res 1617:7-17|
|Schafer, Dorothy P; Lehrman, Emily K; Heller, Christopher T et al. (2014) An engulfment assay: a protocol to assess interactions between CNS phagocytes and neurons. J Vis Exp :|
|Risher, W Christopher; Patel, Sagar; Kim, Il Hwan et al. (2014) Astrocytes refine cortical connectivity at dendritic spines. Elife 3:|
|Schafer, Dorothy P; Stevens, Beth (2013) Phagocytic glial cells: sculpting synaptic circuits in the developing nervous system. Curr Opin Neurobiol 23:1034-40|
Showing the most recent 10 out of 13 publications