Fig. 6: Rare codons in mGluR, an Orb2-regulated mRNA, impact mating behavior.
From: Orb2 enables rare-codon-enriched mRNA expression during Drosophila neuron differentiation
A, C Schematic showing mGluR codon-modified transgenes tagged with a FLAG tag and a Vhh05 nanobody antigen for protein visualization. mGluRendo has the endogenous coding sequence, with a CAI of 0.613 (Fig. 6A), while mGluRcom has the codon optimized coding sequence, with a CAI of 0.985 (Fig. 6B), and mGluRcombs has Orb2 binding sites added back in, for a CAI of 0.945 (Fig. 6C). D Vhh05 protein quantification for mGluRendo heterozygote, mGluR com heterozygote, or mGluRcombs heterozygote adult male brains. Not significant by One-way ANOVA. Mean ± SEM plotted, three biological replicates conducted. Precise N values provided in source data file. E Vhh05 protein quantification of WL3 brains for mGluRendo heterozygotes, mGluRcom heterozygotes, or mGluRcombs heterozygotes. One-way ANOVA followed by Tukey’s multiple comparisons test performed: mGluRendo/+ vs mGluRcom/+ p = 0.0241; mGluRendo/+ vs mGluRcombs/+ p = 0.0442. Mean ± SEM plotted, three biological replicates conducted. Precise N values provided in source data file. F Measurement of time that control or transgenic mGluR heterozygote males spent courting control females in a ten-minute time frame (=courtship index). One-way ANOVA followed by Dunnett’s multiple comparisons test performed. Precise N values provided in source data file, 5 biological replicates conducted. Control vs mGluRcom/+ p = 0.0273; control vs mGluRcombs/+ p = 0.0128. Minima, maxima, centers and bounds of box and whiskers in source data file. G Vhh05 protein quantification of WL3 brains for mGluRendo heterozygotes, mGluRcom heterozygotes, mGluRcombs heterozygotes, each with or without homozygous orb2 deletion. One-way ANOVA followed by Å Ãdák’s multiple comparisons test. mGluRendo/+ vs mGluRcom/+ p = 0.021; mGluRendo/+ vs mGluRcombs/+ p = 0.0432; mGluRendo/+ vs mGluRendo/orb2Δ p = 0.0345; mGluRendo/orb2Δ vs mGluRcom/orb2Δ and mGluRendo/orb2Δ vs mGluRcombs/orb2Δ p < 0.0001. Three replicates conducted, precise N values provided in source data file. H Plot of fold change in Vhh05 protein expression with and without orb2 deletion (full data presented in G). Ratio of Vhh05 expression in orb2 deletion WL3 brains to average of brains with wild-type orb2. One-way ANOVA followed by Tukey’s multiple comparisons test. mGluRendo vs mGluRcom p = 0.0020; mGluRendo vs mGluRcombs p = 0.0089.
