Multiple conserved cell adhesion protein interactions mediate neural wiring of a sensory circuit in C. elegans
- Albert Einstein College of Medicine, United States
Figures
Figure 1 with 3 supplements
Cell adhesion protein genes casy-1 and rig-6 are required for axon fasciculation.
(A) Schematic of the position of cell bodies and axons of three neurons HOA (red), AVG (blue) and a pair of PHC (green) in the male (ventral view). The dashed box indicates the axons and HOA …
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Figure 1—source data 1
Source data for Figure 1, Figure 1—figure supplement 1, and Figure 1—figure supplement 2.
- https://doi.org/10.7554/eLife.29257.006
Figure 1—figure supplement 1
Visualizing HOA presynapses.
(A) Lateral and ventral views of the tail region of animals carrying eat-4pA::gfp (green) and eat-4pA::mcherry::RAB-3 (red). The eat-4pA is a fragment of eat-4 promoter and drives expression only in …
Figure 1—figure supplement 2
Extra branching of HOA in sax-3(ky123) mutants.
(A) HOA axon of wild type or sax-3(ky123) mutants. Arrowheads indicate extra branches of HOA. Scale bar, 20 μm. Percentage of animals with extra branches of HOA was measured and compared to wild …
Figure 1—figure supplement 3
Phenotypes of rig-6(gk438569) mutants.
(A) Distribution of a mCherry-tagged presynaptic marker RAB-3 in HOA axon of rig-6(gk438569) mutants. Arrowheads indicate gaps between the presynaptic puncta. Scale bar, 20 μm. (B) Number of …
Figure 2 with 2 supplements
Axon fasciculation defects of casy-1 and rig-6 are distinguishable.
(A–D) The axons of the three neurons were individually labeled with wCherry (HOA; red), TagBFP (AVG; blue), and GFP (PHC; green) in wild type (A), casy-1(tm718) (B), rig-6(ok1589) (C), or casy-1;rig-…
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Figure 2—source data 1
Source data for Figure 2 and Figure 2—figure supplement 2.
- https://doi.org/10.7554/eLife.29257.010
Figure 2—figure supplement 1
Comparison of PHCL-PHCR defasciculation in electron micrographs and fluorescence images.
(A) Schematic of axon fasciculation between PHCL and PHCR neurons reconstructed from electron micrographs of an adult male N2Y (from section #13883 to #14249). Note that fasciculation of these axons …
Figure 2—figure supplement 2
Phenotypes of rig-6(gk438569) mutants.
(A) Images of axon placement of HOA, AVG and PHC in rig-6(gk438569) mutants. Axon fasciculation between each neuronal pair in the dashed box region and schematic of axon fasciculation are shown on …
Figure 3
CASY-1 and RIG-6 act in postsynaptic AVG for axon fasciculation.
(A and B) Percentage of axon-axon contact between each neuronal pair in animals expressing a full-length casy-1 (A) or rig-6 cDNA (B) in postsynaptic AVG or presynaptic HOA or PHC in a respective …
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Figure 3—source data 1
Source data for Figure 3.
- https://doi.org/10.7554/eLife.29257.012
Figure 4
Structure-function analysis for CASY-1 and RIG-6.
(A and C) Domain structure of mRFP::CASY-1 and its deletion constructs (A) or of YFP::RIG-6 and its deletion constructs (C) with the summary of rescue activity when expressed in AVG of casy-1 or rig-…
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Figure 4—source data 1
Source data for Figure 4.
- https://doi.org/10.7554/eLife.29257.014
Figure 5 with 2 supplements
BAM-2 interacts with CASY-1.
(A) Distribution of a mCherry-tagged presynaptic marker RAB-3 in HOA axon of wild type or bam-2(cy6) mutants. Arrowheads indicate gaps between the presynaptic puncta. (B) Number of mCherry::RAB-3 …
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Figure 5—source data 1
Source data for Figure 5, Figure 5—figure supplement 1, and Figure 5—figure supplement 2.
- https://doi.org/10.7554/eLife.29257.018
Figure 5—figure supplement 1
HOA presynaptic puncta phenotypes of mutants for cell adhesion genes.
(A) Distribution of a mCherry-tagged presynaptic marker RAB-3 in HOA axon of mutants for cell adhesion genes expressed in HOA (red box) or PHC (green box). Expression pattern of these genes is shown …
Figure 5—figure supplement 2
NRX-1 does not interact with CASY-1.
(A) Distribution of a mCherry-tagged presynaptic marker RAB-3 in HOA axon of a hypomorphic nrx-1(ok1649) and a null nrx-1(wy778) mutants. Scale bar, 20 μm. (B) Number of mCherry::RAB-3 puncta in …
Figure 6 with 1 supplement
SAX-7 interacts with RIG-6.
(A) Distribution of a mCherry-tagged presynaptic marker RAB-3 in HOA axon of wild type or sax-7(nj48) mutants. Arrowheads indicate gaps between the presynaptic puncta. (B) Number of mCherry::RAB-3 …
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Figure 6—source data 1
Source data for Figure 6.
- https://doi.org/10.7554/eLife.29257.021
Figure 6—figure supplement 1
sax-7 is expressed in PHC, but not in HOA or AVG.
A fosmid-based reporter line carrying sax-7::gfp was crossed with animals carrying eat-4fosmid::mCherry or unc-47fosmid::mCherry and examined for expression in PHC, HOA or AVG in L4 or adult males. …
Figure 7
Cell adhesion genes required for male mating behavior.
Vulva location efficiency (A) or response efficiency (B) in the indicated mutant males was measured and compared to wild type males. For rescue experiments, the same cell-autonomous rescuing …
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Figure 7—source data 1
Source data for Figure 7.
- https://doi.org/10.7554/eLife.29257.023
Figure 8
Model for function of cell adhesion protein interactions.
(A) Three pathways additively act to achieve the wild type level of HOA-AVG axon fasciculation. The median percentages of HOA-AVG axon fasciculation in mutant and/or PHC ablation studies were used …
Author response image 1
Additional files
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Supplementary file 1
Neighborhood and synapse in the HOA-AVG-PHC circuit
- https://doi.org/10.7554/eLife.29257.025
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Supplementary file 2
C. elegans strains, molecular cloning and transgenes
- https://doi.org/10.7554/eLife.29257.026
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Transparent reporting form
- https://doi.org/10.7554/eLife.29257.027
