Synergistic interactions in <i>Drosophila</i> between <i>shibire</i> and <i>alpha spectrin</i>, the orthologues of ASD-candidate genes from a <i>de novo</i> gain CNV 12235_chr9_gain_129907917_l.

Abstract

<p><b>A.</b> The Locus of the CNV with mapped <i>Drosophila</i> orthologues (Target, green; control, red). <b>B.</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 <i>w</i>^<i>1118</i> controls. However, <i>Shibire</i> and <i>alpha-spectrin</i> 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 <i>Su(P)</i> (using <i>Su(P)</i>^{<i>EY13245</i>}) and <i>CG14104</i> (using <i>CG14104</i>^{<i>f07593</i>}) selected from genes found within CNV gave no significant NMJ phenotype singularly or when crossed to form transheterozygotes with candidate genes. <b>C.</b> Circadian rhythm analysis of candidate genes. Negative controls and candidate gene orthologue overexpression of <i>alpha-spectrin</i> displayed normal light/dark differences in sleeping patterns singularly or when crossed. However, <i>Shibire</i> overexpression, and co-overexpression with <i>alpha-spectrin</i> lost the dark bias, and displayed no significant difference between light/dark sleeping patterns (<b>t</b>).</p