Fig. 5

Disorganisation of axonal MTs upon loss of different MT regulators in Drosophila primary neurons. a Normal neuron (wild-type, wt) with soma (asterisk), axon shaft (curved arrow) and growth cone (tip of most distal MT indicated by arrow head). b Eb1⁵ mutant neuron where the area of MT disorganisation is framed by a red stippled box and shown as close-up on the right. c-e Similar close-ups shown for Efa6^GX6[w-], Khc²⁷ and shot³ mutant neurons. Note that the four mutated factors perform fundamentally different molecular functions, with Eb1 being a MT plus-end binder ('8' in Fig. 3), Efa6 a cortical collapse factor ('4' in Fig. 3), Khc a kinesin-1 motor protein ('A-E' in Fig. 3) and Shot a multi-functional cross-linker ('1-3, 5, 11' in Fig. 3). All neurons were derived from wild-type or homozygous mutant embryos, mechanically and chemically dissociated, kept for 7 days in pre-culture in a centrifuge tube to deplete any maternal gene product, mechanically and chemically dissociated again, cultured on concanavalin A-coated glass coverslips for 1day at 21°C, fixed and stained with anti-α-tubulin (DM1A, Sigma; procedures detailed elsewhere: [78]); images were taken by A.V. using STED (stimulated emission depletion) microscopy. Scale bar in A represents 10 μm for the two neurons and 4 μm in close-ups