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. 2020 Jul 1;9:e55775. doi: 10.7554/eLife.55775

Figure 1. Heterozygous ASD gene mutations do not affect baseline transmission or PHP.

(A) Schematic of the Drosophila locus for CHD8, ASH1L, CHD2, WDFY3 and RIMS1 with gene disruptions indicated. (B) Representative EPSP and mEPSP traces for indicated genotypes (+ / - PhTx for each genotype, left traces and right traces respectively) (C–D) Quantification of mEPSP amplitude (C) and EPSP amplitude (D) in the absence and presence of PhTx (open and filled bars respectively). (E) The percent change of mEPSP and quantal content as indicated, comparing the presence and absence of PhTx for each genotype with Student’s t-test (two tail), *p<0.05, **p<0.01. Sample sizes for data reported (C–E) are as follows (n reported for each genotype -/+ PhTx): wild type: n = 36/47; CHD8/+: n = 7/8; ASH1L/+: n = 15/25; WDFY3/+: n = 8/7; CHD2/+: n = 8/19; RIMS1/+: n = 20/30. (F–H) Scatter plots of quantal content (y axis) versus mEPSP amplitude (x axis) for wild type (left), RIMS1/+ mutant (middle) and the CHD8/+; RIMS1/+ double heterozygous mutant. Each symbol represents an individual muscle recording. Inset: representative traces (+ / - PhTx). Exponential data fit (black line, R2-value inset, calculated based on a linear fit). Dashed lines encompass 95% of all data (absent in (H) for clarity). Below each graph (F–H), boxes display percent PHP (+ / - PhTx for each genotype), statistical values compared to baseline (H).

Figure 1.

Figure 1—figure supplement 1. Patch-Seq analysis of gene expression in type 1b and type 1 s motoneurons.

Figure 1—figure supplement 1.

(A) Image of the larval central nervous system with expression of UAS-CD8-GFP driven by MN1-GAL4. Inset, a rhodamine filled patch electrode targets a single identified motoneuron for excision and sequencing (see Materials and methods). (B) Differential gene expression analysis for two different experiments comparing MN1b (three biological replicates each). Most data rely on unity as expected. (C) Comparison of gene expression for type 1b and type 1 s motoneurons. (D) Expression analysis of ASD gene orthologs in type 1b and type 1 s motoneurons in the third larval instar taken from the patch seq data. Expression is normalized to the well-established, motoneuron-expressed transcription factor mothers against decapentaplegic (mad). As confirmation of predicted gene expression we note the absence of expression for glial cells missing (gcm).

Figure 1—figure supplement 2. Double-heterozygous gene mutation combinations impair homeostatic plasticity.

Figure 1—figure supplement 2.

(A–D) Scatter plots of quantal content (y axis) versus mEPSP amplitude (x axis) for A) wild type; B) ASH1L/+, RIMS1/+ double heterozygous mutant (red) and ASH1L/+ heterozygous mutant (grey); C) CHD2/+; RIMS1/+ double heterozygous mutant (red) and CHD2/+ heterozygous mutant (grey); D) WDFY3/+; RIMS1/+ double heterozygous mutant (red) and WDFY3/+ heterozygous mutant (grey). Each symbol represents an individual muscle recording. Exponential and line data fits (straight line, R2-value inset). Boxes show statistics for curve fits and percent PHP expression (plus/minus PhTx). P-values within boxes report the statistical significance of PHP over genotypic baseline. P-values outside boxed compare PHP expression between genotypes.