Measurements were performed using the NIH Image J software. Values were normalized to values obtained for the control group for each litter. E13 rat dorsal spinal cord explants were dissected and embedded in three-dimensional collagen matrices as described (Charron et al., 2003) and cultured in F12:DMEM (1:1), 10% heat-inactivated horse serum, 40 mM glucose, 2 mM glutaMAX, 100 μg/ml streptomycin sulfate, and 100 U/ml penicillin for 16 hr. Where indicated, Netrin-1 (50 or 100 ng/ml) or VEGF (10, 50, or 100 ng/ml) were added to the medium. Commissural axons

were detected by TAG-1 immunostaining and the total length of axon bundles per explant (for outgrowth) was quantified as described previously (Charron et al., 2003). S3I-201 clinical trial We thank D. Schmucker and L. Moons for helpful advice and discussions, A. McMahon (Harvard University) for providing the Wnt1-Cre mouse line to A.C., A. Nagy for providing the VEGFLacZ mice, D. Anderson for providing the VEGFlox/lox mice, and C. Henderson for providing the Sema3E probe for ISH. The authors also thank N. Dai, M. De Mol, A. Manderveld, B. Vanwetswinkel, K. Peeters, L. Goddé, A. Bouché, P. Vanwesemael, J. Van Dijck, S. Morin, and P.T. Yam for assistance. check details This study was supported by “Long-term structural Methusalem funding by the Flemish Government,” the Fund for Scientific Research-Flemish Government (FWO) (G.0319.07, G.0677.09,

G.02010.07, G.0676.09, 1.5.210.10.N.00 [Krediet aan navorsers]), Concerted Research Activities K.U. Leuven (GOA/2006/11), and the Belgian Science Policy (IUAP-P6/20 and IUAP-P6/30), the Association Française contre les myopathies (AFM), Geneeskundige stichting Koningin Elisabeth, and MND/A grant 70/130. C.R.A. is postdoctoral over fellow of the FWO (1.2.545.09.N.00, V.4.332.10.N.01). C.C. is a fellow of the Flemish Institute for the promotion of scientific research (IWT), Belgium. I.S. is a postdoctoral fellow of the European Union Seventh framework program. C.L. is supported by an EMBO long-term postdoctoral fellowship. A.C. is supported by grants from the “Fondation pour la recherche médicale” (programme

Equipe FRM) and the Agence Nationale de la Recherche (ANR-08-MNPS-030-01). F.C. is a Fonds de la recherche en santé du Québec (FRSQ) Scientist. Work performed in the Charron laboratory was supported by an operating grant from the Canadian Institutes of Health Research (CIHR). “
“Alcohol can diminish feelings of anxiety and stress, boost mood, enhance sociability, and induce sleep. Unfortunately, it has also been classified as the most harmful psychoactive drug we abuse (Nutt et al., 2010). Alcohol abuse is widespread, and alcohol use disorders (AUDs) are a debilitating individual and societal problem, diagnosed in over 76 million people worldwide (WHO, 2004). Genetic predispositions have a strong influence on AUDs.