New X-chromosomal interactors of dFMRP regulate axonal and synaptic morphology of brain neurons in Drosophila melanogaster

Figures & data

Figure 1. Wing phenotypes of flies with different genotypes: wild-type wing (A); overexpression of dfmr1 causes ‘notched’ wing phenotype (control) (B); enhancer wing phenotype (C); suppressor wing phenotype (D); overexpression of rok (E); overexpression of sgg (F).

Figure 2. Confocal images of LNv's from adult half-brains with different genotypes: wild type (A); heterozygotes for the dfmr1-null mutation (B); heterozygotes peb^[hnt-E8]//+ (C); heterozygotes peb^[hnt-E8]//+ (D); double heterozygotes peb^[hnt-E8]//+; Fmr1^Δ113M//+ (E); heterozygotes rok¹//+ (F); double heterozygotes rok¹//+; Fmr1^Δ113M//+ (G); heterozygotes sgg^G0263//+ (H); double heterozygotes sgg^G0263//+; Fmr1^Δ113M//+ (I); heterozygotes ras^G0380b//+ (J); double heterozygotes ras^G0380b//+; Fmr1^Δ113M//+ (K). The target-area coverage for each genotype (L); Mean fluorescence intensity (M). All brains are shown at 40× magnification. Error bars indicate SD.

Table 1. Ectopic branching and defasciculated axonal bundles of Drosophila LNv's.

	WT	peb^[^hnt-E^8]//+	rok^1//+	sgg^G^0263//+	ras^G^0380b //+
Ectopic axonal branching (per cent of total half-brains)	0%	60%	0%	13%	0%
	0 (6)	6 (10)	0 (10)	2 (15)	0 (10)
	n = 6	n = 10	n = 10	n = 15	n = 10
Defasciculated s-LNv-axon bundle (per cent of total half-brains)	0%	60%	40%	33%	40%
	0 (6)	6 (10)	4 (10)	6 (15)	4 (10)
	n = 6	n = 10	n = 10	n = 15	n = 10
Defasiculated bundle of POT (per cent of total half-brains)	0%	50%	10%	40%	80%
	0 (6)	5 (10)	1 (10)	6 (15)	8 (10)
	n = 6	n = 10	n = 10	n = 15	n = 10

Figure 3. Western blot analysis. Expression levels in wild-type, dfmr1-null mutants and in the case of dfmr1 overexpression in the brains of: Peb/HNT – about 209 kDa (A); Rok – about 160 kDa (B); Shaggy/GSK3β (C) for each isoform: SGGY (about 56 kDa), SGG10 (about 58 kDa) and SGG39 (about 68 kDa); Ras – about 60 kDa (D). Expression levels of dFMRP (about 85 kDa) in embryos hemizygous for peb^[hnt-E8], rok¹, sgg^G0263 or ras^G0380b (E) (p > 0.05; n = 3). Quantification of each protein expression from Western blot analysis is shown on the right side. Error bars indicate SD.