The mean of YFP/CFP ratios for VA8 and VB9 during these periods was considered the averaged activity for VA8 and VB9 for each animal during the indicated mode of locomotion. The difference between the VB9 and VA8 activity level in each animal was normalized by [VB9 − VA8]/[VB9 + VA8]. For correlation analysis, VA8 and VB9 transients of each sample were corrected for photobleaching by dividing fitted linear regression line, normalized by

mean and SD. For correlation analyses of VA8 and VB9 activity change during the transition of directions, eight VA8/VB9 imaging traces were used for correlation analyses. Pearson’s correlation coefficient was calculated by R. Selleck JAK inhibitor Detailed procedures for curvature analysis, automated movement analysis, electrophysiology, molecular biology, neuron silencing, ablation and chemical synapse inactivation, immunofluorescent staining, and statistical analysis are provided in Supplemental Experimental Procedures. We thank H. Li and Y. Wang for technical support; A.V. Maricq for akIs11, Z.W. Wang for UNC-9 antisera, C. Bargmann for tetanus toxin cDNA, S. Lockery for exchanging unpublished results, and M. Zhang and H. Suzuki for advice on calcium imaging. We are in debt to L. Avery, C. Bargmann,

J.-L. Bessereau, J. Culotti, C.-Y. Ho, A. Kania, J. Richmond, Q. Wen, J. Woodgett, and anonymous reviewers for critical reading and comments on this manuscript. M. Po was a recipient of a Natural Sciences and Quisinostat supplier Engineering Research Council of Canada scholarship. We thank the EJLB foundation, the Canadian Institute of Health Research and the Samuel Lunenfeld Research Institute for supporting this project. “
“Synaptic transmission relies on the fusion of synaptic vesicles (SVs) with the presynaptic plasma membrane (exocytosis) to release neurotransmitters. Ketanserin After exocytosis, excess plasma membrane resulting from the addition of SV membrane is rapidly internalized by compensatory endocytosis and used to generate new SVs. Proper nervous system function relies critically on

the efficiency of this membrane-recycling traffic. Clathrin-mediated endocytosis is a major pathway for SV recycling (Dittman and Ryan, 2009 and Heuser and Reese, 1973). In this process, nucleation and growth of the clathrin coat helps gather proteins to be internalized and generates and stabilizes the bilayer curvature required for the formation of the endocytic bud. PI(4,5)P2, a phosphoinositide selectively enriched in the plasma membrane, plays a key role in the recruitment and assembly of the endocytic clathrin adaptors, which, in turn, recruit and promote the assembly of clathrin (Di Paolo and De Camilli, 2006). After a deeply invaginated clathrin-coated pit (CCP) is generated, it undergoes fission with the help of the GTPase dynamin (Ferguson et al., 2007 and Raimondi et al., 2011) and then rapidly loses its coat.