Regulation of neuronal axon specification by glia-neuron gap junctions in C. elegans

  1. Lingfeng Meng
  2. Albert Zhang
  3. Yishi Jin
  4. Dong Yan  Is a corresponding author
  1. Duke University Medical Center, United States
  2. Duke Medical Center, United States
  3. Howard Hughes Medical Institute, University of California, United States
  4. School of Medicine, University of California, United States
7 figures and 2 additional files

Figures

Figure 1 with 2 supplements
unc-1 is required for RME axon specification.

(A) A cartoon picture and (B) GFP images show the morphology of RME neurons. Cell bodies of four RME neurons, labeled as D (RME dorsal), V (RME ventral), L (RME left) and R (RME right), form a …

https://doi.org/10.7554/eLife.19510.002
Figure 1—figure supplement 1
ju1057 is a loss-of-function allele of unc-1.

(A) Quantification of the fractions of SNB-1::GFP signals in RME D/V neurites (green), axonal boutons (blue) and cell bodies (red) in control and unc-1(lf) animals. The total intensity of SNB-1::GFP …

https://doi.org/10.7554/eLife.19510.003
Figure 1—figure supplement 2
unc-1(lf) affects RME axon specification.

(A) Images showed that loss-of-function in unc-1(e580) (without unc-30(ju32) background) caused the misaccumulation of SNB-1::GFP puncta in both RME nerve ring processes and RME D neurites, Scar …

https://doi.org/10.7554/eLife.19510.004
Figure 2 with 1 supplement
Gap junctions between GLR cells and RME neurons are required for RME axon specification.

(A) Quantification of SNB-1::GFP puncta distribution defects in unc-1(lf) mutants and rescue transgenic strains. Promoters used to drive expression of unc-1 cDNA are: Pdpy-30 in all tissues; Punc-33

https://doi.org/10.7554/eLife.19510.005
Figure 2—figure supplement 1
GLR cells and RME neurons

(A) A confocal image shows the relative positions of GLR cells (Pink) and RME neurons (Green). GLR cells were labeled by expression of Pgly-18::mCherry, and RME neurons were labeled by expression of …

https://doi.org/10.7554/eLife.19510.006
Figure 3 with 1 supplement
unc-1 regulates microtubule polarity in RME D/V dendrites.

(A) Time kymographs generated from a 40-s movie of EBP-2::GFP dynamics in control, unc-1(e580) and unc-7 unc-9 animals. (B) Quantification of percentage of orientation in RME D/V neurites in control …

https://doi.org/10.7554/eLife.19510.007
Figure 3—figure supplement 1
unc-1(lf) does not change the amount of polymerized microtubules in RME D/V dendrites.

Confocal images (A) and quantification (B) of EMTB::GFP signals in RME D/V neurites. n = 15. Data are shown as mean ± SD. Student test, NS: no significant difference. Pink-colored neurons are AIY …

https://doi.org/10.7554/eLife.19510.008
The CDK-5 pathway is regulated by GLR-RME gap junctions in RME axon specification

(A) Loss-of-function of cdk-5 induces microtubule polarity defects in RME D/V neurites to a similar degree as those in unc-1(lf) animals. cdk-5: n = 81. Values that differ significantly from wild …

https://doi.org/10.7554/eLife.19510.009
GLR-RME gap junctions regulate the CDK-5 pathway through calcium and calpain.

(A) Gel images and (B) quantification results show that calcium antagonist BAPTA-AM and Calpain inhibitor PD150606 suppress the cleavage of CDKA-1/p35. Three independent experiments were performed, …

https://doi.org/10.7554/eLife.19510.010
slo-1(lf) suppresses unc-1(lf) phenotypes through the CDK-5 pathway.

(A) Quantification of SNB-1::GFP distribution defects in unc-1(ju1057), slo-1 and unc-1(ju1057);slo-1. (B) Quantification of SNB-1::GFP distribution defects in animals expressing calbindin D28K …

https://doi.org/10.7554/eLife.19510.011
A model for GLR-RME gap junction regulated RME axon specification.

At the early developmental stage, calcium influxes from GLR cells to RME neurons through gap junctions to induce activation of the CDK-5 pathway, which guides microtubule assembly and axon …

https://doi.org/10.7554/eLife.19510.012

Additional files

Supplementay file 1

Supplemental Table 1, Strain list.

https://doi.org/10.7554/eLife.19510.013
Supplementay file 2

Supplemental Table 2, Plasmid list.

https://doi.org/10.7554/eLife.19510.014

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