FIGURE

Fig. 3

ID
ZDB-FIG-180105-16
Publication
Lee et al., 2017 - The Kinesin Adaptor Calsyntenin-1 Organizes Microtubule Polarity and Regulates Dynamics during Sensory Axon Arbor Development
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Fig. 3

Clstn-1 loss-of-function (lof) disrupts MT polarity in sensory neuron peripheral axons. (A) In situ hybridization showing clstn-1 mRNA expression in 24 hpf wild type or Clstn-1−/− embryos. (B) qPCR results showing clstn-1 mRNA expression levels normalized to an EF1α positive control (wild type mRNA = −4.681, n = 3 biological replicates of 50 embryos; Clstn-1−/− mRNA = −7.616, n = 2 biological replicates of 50 embryos; *p = 0.014 student's t-test). (C) RB neuron labeled with EB3-GFP in Clstn-1 MO injected embryo. Retrograde comets appear frequently in Clstn-1 lof peripheral axons. Scale bar is 10 μm. (C') Kymograph of the white boxed region in C shows several retrograde comets highlighted in red. Scale bars are 5 μm (y) and 1 min (x). (C”) Time-lapse sequences of the yellow boxed area show a retrograde comet traveling between two branch points (red arrowheads). Time is shown in min:sec. (D) Clstn-1 lof embryos have a higher percentage of retrograde comets than wild type (WT). (WT peripheral = 3.69%, n = 18 neurons; Clstn-1 MO peripheral = 9.29%, n = 18 neurons, significant via student's t-test comparison to wild type, p = 0.012, but not with Dunnett's post-test after one-way ANOVA p = 0.086; Clstn-1−/− peripheral = 13.55%, n = 17 neurons, ***p = 0.0009 student's t-test, **p = 0.0017 Dunnett's post-test; p = 0.0034 one-way ANOVA). In central axons, there is no significant difference in percentage of retrograde comets between wild type and Cltn-1 lof. (WT central = 9.11%, n = 25 neurons; Clstn-1 MO central = 12.60%, n = 18 neurons, p = 0.27 student's t-test, p = 0.49 Dunnett's post-test; Clstn-1−/− central = 12.10%, n = 16 neurons, p = 0.39 student's t-test, p = 0.61 Dunnett's post-test; one-way ANOVA p = 0.54). (E) Clstn-1−/− neurons have a higher frequency of retrograde comets in peripheral axons than wild type (mean WT peripheral frequency = 0.0039 comets/μm/min, n = 81 axon segments; mean Clstn-1 MO peripheral frequency = 0.0043 comets/μm/min, n = 63 axon segments, p = 0.90 Dunnett's post-test; mean Clstn-1−/− peripheral frequency = 0.012 comets/μm/min, n = 47 axon segments, ***p = 0.0007 Dunnett's post-test; one-way ANOVA ***p = 0.0006). (F) In Clstn-1 lof neurons anterograde comets in central axons travel faster than those in peripheral axons, similar to wild type, (mean Clstn-1 MO peripheral velocity = 5.23 μm/min, n = 63 axon segments, mean Clstn-1 MO central velocity = 7.05 μm/min, n = 26 axon segments, ****p < 0.0001 student's t-test; mean Clstn-1−/− peripheral velocity = 5.53 μm/min, n = 48 axon segments, mean Clstn-1−/− central velocity = 8.74 μm/min, n = 24 axon segments, ****p < 0.0001 student's t-test, wild type velocity data repeated from Figure 1 show here again for comparison). Clstn-1−/− anterograde comets in central axons travel faster than their wild type counterparts (WT vs. Clstn-1−/− mean central velocity *p = 0.031 Dunnett's post-test, WT vs. Clstn-1 MO mean central velocity: p = 0.91 Dunnett's post-test, *p = 0.048 one-way ANOVA).

Expression Data
Gene:
Fish:
Anatomical Terms:
Stage: Prim-5

Expression Detail
Antibody Labeling
Phenotype Data
Fish:
Knockdown Reagent:
Observed In:
Stage Range: 14-19 somites to Prim-5

Phenotype Detail
Acknowledgments
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Front. Cell. Neurosci.