astrocytic ca2+ waves guide cns growth cones to remote regions of neuronal activity星形ca2 +波引导中枢神经生长锥神经活动的偏远地区.pdfVIP

Astrocytic Ca2+ Waves Guide CNS Growth Cones to Remote Regions of Neuronal Activity Johanna Hung, Michael A. Colicos* Department of Physiology and Biophysics and the Hotchkiss Brain Institute, University of Calgary, Calgary, Alberta, Canada Abstract Activity plays a critical role in network formation during developmental, experience-dependent, and injury related remodeling. Here we report a mechanism by which axon trajectory can be altered in response to remote neuronal activity. Using photoconductive stimulation to trigger high frequency action potentials in rat hippocampal neurons in vitro, we find that activity functions as an attractive cue for growth cones in the local environment. The underlying guidance mechanism involves astrocyte Ca2+ waves, as the connexin-43 antagonist carbenoxolone abolishes the attraction when activity is initiated at a distance greater than 120 mm. The asymmetric growth cone filopodia extension that precedes turning can be blocked with CNQX (10 mM), but not with the ATP and adenosine receptor antagonists suramin (100 mM) and alloxazine (4 mM), suggesting non-NMDA glutamate receptors on the growth cone mediate the interaction with astrocytes. These results define a potential long-range signalling pathway for activity-dependent axon guidance in which growth cones turn towards directional, temporally coordinated astrocyte Ca2+ waves that are triggered by neuronal activity. To assess the viability of the guidance effect in an injury paradigm, we performed the assay in the presence of conditioned media from lipopolysaccharide (LPS) activated purified microglial cultures, as well as directly activating the glia present in our co- cultures. Growth cone attraction was not inhibited under these conditions, suggesting this mechanism could be used t