Fig. 5
From: Sphingolipid-dependent Dscam sorting regulates axon segregation
Neuronal polarity is not affected in SPT mutant. a–f Distribution of various dendritic (Apc2::GFP: (a, d)) (nod::GFP: (b, e)) (DenMark: (c, f)), synaptic (Syt::GFP: (c, f)) and axonal marker (FasII: (a–f)) in lacek05/ + (a–c) and lace2/lacek05 (d–f) expressed using OK107-Gal4 (exception: α/β specific Gal4 (R65G04-Gal4) for DenMark). While images a–f represent the axonal half of the MB, the corresponding somato-dendritic area is represented in right upper insets. The white dotted rectangle represents the indicated GFP-tagged marker distribution in the axons, zoomed in and shown in the right lower inset. While the dendritic markers do not show a change in the somato-dendritic distribution (a, b, d, e), the synaptic marker Syt::GFP shows the formation of aggregates in lace mutants ((c, f), arrowhead). The axonal distribution of FasII remains unchanged (a–f). n(a) = 38, n(b) = 34, n(c) = 28, n(d) = 36, n(e) = 18, n(f) = 38. g–o Somato-dendritic region of the indicated SPT mutant genotypes without (g–l) or with (j–o) Dscam[TM1]::GFP expression and anti-FasII staining. m, n, o represent processed (j, k, l) images for visualization of colocalization (White) of Dscam[TM1] (Green) and FasII (Red) aggregates. FasII shows the formation of aggregates in the somato-dendritic compartment in strong SPT mutants ((h, i) arrowheads). In addition, a smooth membranous shift of FasII into the calyx was observed (H, arrow). Co-labeling of FasII with Dscam[TM1]::GFP shows that both molecules form mostly separate aggregates in the somato-dendritic region though some colocalization was also observed ((k, l, n, o) hollow arrowheads). n(g) = 64, n(h) = 62, n(i) = 38, n(j, m) = 30, n(k, n) = 48, n(l, o) = 32. p–w Distribution of dendritic (Dscam[TM1]::GFP, (p, t)) (Apc2::GFP, (r, v)), axonal (Dscam[TM2]::GFP, (q, u)), and synaptic (Syt::GFP, (s, w)) markers in third instar larval MBs in lace2/ + (p–s) and lace2/lacek05 (t–w) driven with 201Y-Gal4. No change in the distribution of the markers was observed. Further, the AIS in the larval γ neurons seemed unperturbed in lace mutants (r, v, s, w). n(p) = 26, n(q) = 24, n(r) = 20, n(s) = 20, n(t) = 42, n(u) = 12, n(v) = 10, n(w) = 16. a–e, g–w Green: GFP, Red: FasII, Blue: N-Cad. Scale Bar: 25 μm
