Synergistic Interactions between Drosophila Orthologues of Genes Spanned by De Novo Human CNVs Support Multiple-Hit Models of Autism
Fig 5
Synergistic interactions in Drosophila between shibire and alpha spectrin, the orthologues of ASD-candidate genes from a de novo gain CNV 12235_chr9_gain_129907917_l.
A. The Locus of the CNV with mapped Drosophila orthologues (Target, green; control, red). B. Synaptic alterations were characterised by NMJ type IB bouton number. Individual heterozygous mutants of candidate gene orthologues gave no significant change in NMJ morphology over w1118 controls. However, Shibire and alpha-spectrin double over expressers display reduced bouton numbers (using 1032-GAL4; UAS-Dynamin/UAS-alpha-spectrin; n>20, Kruskal-Wallis test, * P<0.05). Non-candidate gene controls Su(P) (using Su(P)EY13245) and CG14104 (using CG14104f07593) selected from genes found within CNV gave no significant NMJ phenotype singularly or when crossed to form transheterozygotes with candidate genes. C. Circadian rhythm analysis of candidate genes. Negative controls and candidate gene orthologue overexpression of alpha-spectrin displayed normal light/dark differences in sleeping patterns singularly or when crossed. However, Shibire overexpression, and co-overexpression with alpha-spectrin lost the dark bias, and displayed no significant difference between light/dark sleeping patterns (t).