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. 2022 Oct 12;11:e74270. doi: 10.7554/eLife.74270

Figure 9. Microtubule dynamics are increased in klc4uw314 mutants.

(A-A”) Live RB neuron expressing EB3-GFP, scale bar = 10 μm. Magenta box in (A) indicates axon segment selected for kymograph in (A’). (A”) EB3 comets identified as anterogradely (green) or retrogradely (magenta) directed based on their angle. (B) Representative kymographs from peripheral axon segments of wildtype (top) or klc4uw314 (bottom). (C) Quantification of the percentage of retrograde comets out of all comets imaged for both peripheral and central axons. NS by Mann-Whitney test. (D) Peripheral axon comet frequency. ***p=0.0003, Mann-Whitney test. (E) Peripheral axon comet velocity. ***p=0.0004, *p=0.042, Mann-Whitney test. For all data, error bars = SEM. Wild type central N=8 segments, klc4uw314 central N=15 segments, Wild type peripheral N=26 segments, klc4uw314 peripheral N=48 segments. Wild type data from 10 neurons in 7 embryos. Klc4uw314 data from 13 neurons in 8 embryos. All comets in a kymograph were measured.

Figure 9—source data 1. Source Data 1: All measurements of EB3-GFP comets in wild type and klc4uw314 mutants, averaged per measured segment.

Figure 9.

Figure 9—figure supplement 1. Additional quantification of microtubule dynamics in wild type and klc4 mutant axons.

Figure 9—figure supplement 1.

(A) Central axon comet frequency. NS by Mann-Whitney test. (anterograde p=0.4664, retrograde p=0.7268) (B) Central axon comet velocity. *p<0.05, Mann-Whitney test. (C) Average distance traveled by peripheral axon comets. NS by Mann-Whitney test (anterograde p=0.1476, retrograde p=0.9288) (D) Average distance traveled by central axon comets. NS by Mann-Whitney test (anterograde p=0.3738, retrograde p=0.8836) (E) Average comet run time for peripheral axon comets. NS by Mann-Whitney test (***p=0.0001, NS p=0.1389) (F) Average comet run time for central axon comets. NS by Mann-Whitney test (anterograde p=0.3363, retrograde p=0.3196) For all data, error bars = SEM. Wild type central N=8 segments, klc4uw314 central N=15 segments, Wild type peripheral N=26 segments, klc4uw314 peripheral N=48 segments. Wild type data from 10 neurons in 7 embryos. Klc4uw314 data from 13 neurons in 8 embryos.
Figure 9—video 1. An RB neuron expressing EB3-GFP to tag plus ends of microtubules.
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Plus end comets can be seen traveling in a predominantly anterograde direction.