Aberrant information transfer interferes with functional axon regeneration
- Yale University, United States
Figures
Figure 1 with 1 supplement
DA9 axon regeneration and degeneration.
(A) Diagram of DA9 morphology and the laser cutting site. (B) A control adult expressing GFP::RAB-3 and mCherry in DA9. mCherry expression is also observed in the posterior gut in these micrographs. …
Figure 1—figure supplement 1
ric-7(n2657) animals show normal DA9 development, better regeneration and increased degeneration of GABA motor neurons.
(A and B) GFP::RAB-3 and mCherry labeling in DA9 of control (A) and ric-7(n2657) (B) animals (2-day-old adults). Scale bars = 10 µm. (C) Length of the asynaptic region and the synaptic region in …
Figure 2 with 2 supplements
DA9 forms normal and dendritic synapses during regeneration.
(A) Diagram of a regenerated DA9 axon. (B–D) DA9 axonal and synaptic regeneration at different timepoints after axotomy. The white lightning mark indicates the axotomy site. Scale bars = 10 μm. (E) …
Figure 2—figure supplement 1
Axotomy in the DNC enables accurate axon growth following the original path.
(A) DA9 regeneration 48 hr after axotomy in the DNC. The lightning mark indicates the cutting site. The scale bar = 10 μm. (B) DA9 regeneration 48 hr after axotomy in the dorsal-ventral midline of …
Figure 2—figure supplement 2
Regeneration of other presynaptic components.
(A–A’’) UNC-10 localization in intact DA9. Scale bars = 10 μm. (B–B’’) UNC-10 localization in regenerated DA9. Scale bars = 10 μm. (C–C’’) CLA-1S localization in intact DA9. Scale bars = 10 μm. (D–D’…
Figure 3 with 1 supplement
Postsynaptic receptors to DA9 maintain their localization and are aligned with DA9 SV clusters after axotomy.
(A) Labeling of DA9 axon and ACR-12 in GABA motor neurons in intact animals (2d old adults). Scale bars = 10 μm. (B) Labeling of DA9 axon and ACR-12 in GABA motor neurons in axotomized animals. …
Figure 3—figure supplement 1
No change of postsynaptic receptors in body wall muscles after axotomy.
(A–A’’) Muscle ACR-16 localization in intact animals. Note its localization in both the dorsal and ventral nerve cords. Scale bars = 10 μm. (B–B’’) Muscle ACR-16 maintains its localization 48 hr …
Figure 4 with 2 supplements
Aberrant recovery of a single neuron-driven behavior.
(A) Diagram of the optogenetically triggered tail-bending behavior. (B) Images from an example movie showing the dorsal tail-bending behavior. (C–H) Quantification of the tail-bending behavior of …
Figure 4—figure supplement 1
Optogenetically-triggered tail-bending behavior requires Chrimson and ATR and is DA9 specific.
(A) ric-7(n2657) animals expressing mCherry instead of Chrimson supplied with ATR show no dorsal tail bending. Mean ± SEM. The shaded area represents the 5 s stimulation. (B) ric-7(n2657) animals …
Figure 4—figure supplement 2
Optogenetically-triggered tail-bending behavior depends on synaptic transmission.
(A) Tail-bending behavior of intact unc-13(e51) animals (2-day-old adults). (B) Tail-bending behavior of unc-13(e51) animals 48 hr after axotomy. (C) Averaged tail angle during stimulation of both …
Figure 5 with 1 supplement
Simultaneous optogenetic stimulation and calcium imaging reveals rerouted information transfer in the regenerated circuit.
(A–D) Heat maps of calcium signal fold changes (df/f) in intact (A and C) and axotomized animals (B and D) 12 hr or 48 hr after axotomy. White dashed lines delineate the animals’ tails. Scale bars = …
Figure 5—figure supplement 1
Optogenetically induced muscle calcium response requires all-trans retinal and depends on synaptic transmission.
(A) Calcium responses in intact adult animals cultured with all-trans retinal (ATR) (n = 17). (B) No calcium responses in intact adult animals cultured without ATR (n = 24). (C) No calcium responses …
Figure 6 with 1 supplement
Aberrant information transfer is independent of dlk-1 and suppresses behavioral recovery.
(A and B) GFP::RAB-3 and mCherry labeling of DA9 in intact and axotomized ric-7(n2657); dlk-1(ju476) animals 48 hr after axotomy. Scale bars = 10 μm. (C and D) Comparison of length of regenerating …
Figure 6—figure supplement 1
Behavior response of ric-7(n2657); dlk-1(ju476) animals with and without axotomy.
(A) ric-7(n2657); dlk-1(ju476) intact animals show robust dorsal tail bending. The shaded area represents the 5 s stimulation. Mean ± SEM. (B) ric-7(n2657); dlk-1(ju476) axotomized animals show …
Figure 7 with 1 supplement
Dendritic microtubule polarity is maintained after axotomy.
(A) Diagram of regions of EBP-2::GFP imaging in DA9. (B–E) Kymographs of EBP-2 traces in both DA9 dendrite and axon in intact and axotomized animals 12 hr after axotomy. Scale bars are 5 μm and 5 s. …
Figure 7—figure supplement 1
ACR-2, a subunit of the nicotinic acetylcholine receptor, maintains dendritic localization after axotomy.
(A) ACR-2::GFP localizes to the dendrite and soma of DA9 in intact ric-7(n2657) animals. The scale bar = 10 μm. (B) ACR-2::GFP still localizes to the dendrite and soma of DA9 in axotomized ric-7(n265…
Figure 8 with 1 supplement
JNK-1 and dynein-mediated transport mediate SV mislocalization to the dendrite and loss of JNK-1 improves behavioral recovery.
(A–D) GFP::RAB-3 localization 48 hr after axotomy in ric-7(n2657) (A), ric-7(n2657); dhc-1(js319) (B), ric-7(n2657); nud-2(ok949) (C) and ric-7(n2657); jnk-1(gk7) (D) animals. Asterisk indicates …
Figure 8—figure supplement 1
DA9 in dhc-1(js319) animals develops normally and loss of JNK-1 rescues RAB-3 mis-localization in DA9 dendrite 12 hr after axotomy.
(A–B) RAB-3::GFP and mCherry labeling of DA9 in intact ric-7(n2657) and ric-7(n2657); dhc-1(js319) animals (2-day-old adults). Scale bars = 10 μm. (C–F) RAB-3 localization to the dendrite inArxiocn-7…
Additional files
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Transparent reporting form
- https://doi.org/10.7554/eLife.38829.021
