1. Cell Biology
  2. Neuroscience

Hypoxia-inducible factor cell non-autonomously regulates C. elegans stress responses and behavior via a nuclear receptor

  1. Corinne L Pender
  2. H Robert Horvitz  Is a corresponding author
  1. Massachusetts Institute of Technology, United States
https://doi.org/10.7554/eLife.36828
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Cite this article
  1. Corinne L Pender
  2. H Robert Horvitz
(2018)
Hypoxia-inducible factor cell non-autonomously regulates C. elegans stress responses and behavior via a nuclear receptor
eLife 7:e36828.
https://doi.org/10.7554/eLife.36828
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6 figures, 1 table and 3 additional files

Figures

Figure 1 with 2 supplements
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The cytochrome P450 gene cyp-36A1 is an effector of the hypoxia-response pathway that regulates behavior.

(A) Schematic of the screen design. Stages of embryonic development adapted from Ringstad and Horvitz (2008) and Paquin et al. (2016). (B–F) Distribution of stages of eggs newly laid by adult …

https://doi.org/10.7554/eLife.36828.002
Figure 1—figure supplement 1
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A screen for suppressors of the egl-9(lf) egg-laying defect.

(A) Summary of three cloned alleles from a suppressor screen of the egl-9(lf) egg-defect. (B) Gene diagram of hif-1; isoform F38A6.3b is shown. The ia4 deletion allele was generated by deletion …

https://doi.org/10.7554/eLife.36828.003
Figure 1—figure supplement 2
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cyp-36A1(lf) suppresses the egg-laying defect of hypoxia-exposed animals.

(A) Schematic depicting experimental design. (B–E) Distribution of stages of eggs laid by adult hermaphrodites. (B) Stages of eggs laid by wild-type animals grown in normoxia (21% O2). (C) Animals …

https://doi.org/10.7554/eLife.36828.004
Figure 2 with 1 supplement
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CYP-36A1 acts downstream of HIF-1 to regulate gene expression changes and stress responses.

(A) Blue circle: Genes that were at least twofold downregulated in egl-9(lf) mutants. Purple and orange circles: Subset of egl-9(lf)-downregulated genes that were significantly upregulated in egl-9(l…

https://doi.org/10.7554/eLife.36828.005
Figure 2—source data 1

RNA-seq GO enrichment analysis.

Gene ontology enrichment analysis terms for genes more than twofold upregulated in egl-9 mutants and significantly suppressed by hif-1(lf) and cyp-36A1(lf).

https://doi.org/10.7554/eLife.36828.007
Figure 2—figure supplement 1
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Replicate data for survival on Pseudomonas aeruginosa.

Replicate data for Figure 2C. Wild type (n = 150) vs. egl-9(lf) (n = 50), p<0.001; egl-9(lf) vs. cyp-36A1(lf); egl-9(lf) (n = 163), p<0.001; cyp-36A1(lf); egl-9(lf) vs. cyp-36A1(lf); egl-9(lf); nIs …

https://doi.org/10.7554/eLife.36828.006
Figure 3 with 4 supplements
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HIF-1 and CYP-36A1 cell non-autonomously regulate expression of a stress-responsive gene.

(A) PT24B8.5::gfp expression of the indicated genotypes (n = 10 animals per image). Scale bars, 100 μm. (B) Expression of cyp-36A1(+) specifically in neurons, hypoderm, muscle, or intestine of cyp-36…

https://doi.org/10.7554/eLife.36828.008
Figure 3—figure supplement 1
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cyp-36A1 expression in multiple tissues rescues the egg-laying phenotype of cyp-36A1(lf); egl-9(lf).

(A–H) Distribution of stages of eggs laid by adult hermaphrodites of the indicated genotypes. All strains contained the agIs219 (PT24B8.5::gfp) transgene. (A) Stages of eggs laid by wild-type …

https://doi.org/10.7554/eLife.36828.009
Figure 3—figure supplement 2
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cyp-36A1 is expressed in many tissues.

(A–D) Paired fluorescent (left in each panel) and Nomarski (right in each panel) micrographs showing expression of a transcriptional Pcyp-36A1::gfp reporter (nIs682) in an adult worm. Expression was …

https://doi.org/10.7554/eLife.36828.010
Figure 3—figure supplement 3
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Replicate data for PT24B8.5::gfp reporter expression.

Replicate data for Figure 3C. Quantification of n = 10 animals. ***p<0.001 considered significant. ns (p>0.05), not significant (Student’s t-test with Holm-Bonferroni correction).

https://doi.org/10.7554/eLife.36828.011
Figure 3—figure supplement 4
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Replicate data for PT24B8.5::gfp reporter expression.

Replicate data for Figure 3E. Quantification of n = 10 animals. **p<0.01, ***p<0.001 considered significant (Student’s t-test with Holm-Bonferroni correction).

https://doi.org/10.7554/eLife.36828.012
Figure 4 with 2 supplements
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The nuclear hormone receptor NHR-46 acts downstream of CYP-36A1.

(A) nhr-46 gene diagram; isoform C45E5.6b is shown. n6125 and n6126 are in the NHR-46 ligand-binding domain. Scale bar, 100 bases. (B) PT24B8.5::gfp fluorescence of the indicated genotypes (n = 10 …

https://doi.org/10.7554/eLife.36828.013
Figure 4—figure supplement 1
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Replicate data for PT24B8.5::gfp reporter expression.

Replicate data for Figure 4C. Quantification of n = 10 animals. ***p<0.001 considered significant (Student’s t-test with Holm-Bonferroni correction).

https://doi.org/10.7554/eLife.36828.014
Figure 4—figure supplement 2
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Replicate data for survival on Pseudomonas aeruginosa.

Replicate data for Figure 4E. Wild type (n = 34) vs. egl-9(n586) (n = 61), p<0.001; egl-9(lf) vs. cyp-36A1(lf); egl-9(lf) (n = 52), p<0.05; cyp-36A1(lf); egl-9(lf) vs. cyp-36A1(lf); nhr-46(lf); …

https://doi.org/10.7554/eLife.36828.015
Figure 5 with 1 supplement
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NHR-46 acts in different tissues to regulate T24B8.5 expression and egg laying.

(A) Expression of nhr-46(+) in the intestine but not in neurons or muscle rescued the high PT24B8.5::gfp expression in the intestine of cyp-36A1(lf); nhr-46(lf); egl-9(lf) mutants (n = 10 animals …

https://doi.org/10.7554/eLife.36828.016
Figure 5—figure supplement 1
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Replicate data for PT24B8.5::gfp reporter expression.

Replicate data for Figure 5B. Quantification of n = 10 animals. **p<0.01, ***p<0.001 considered significant. ns (p>0.05), not significant (Student’s t-test with Holm-Bonferroni correction).

https://doi.org/10.7554/eLife.36828.017
Figure 6
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Model for the regulation of physiology and behavior by NHR-46 and CYP-36A1.

(A) The genetic pathway in which egl-9 inhibits hif-1, which activates cyp-36A1, which in turn inhibits nhr-46. (B) Model for how CYP-36A1 and NHR-46 function downstream of HIF-1. We suggest that …

https://doi.org/10.7554/eLife.36828.018

Tables

Key resources table
Reagent type
(species) or resource		DesignationSource or referenceIdentifiersAdditional
information
Strain, strain background
(Caenorhabditis elegans)		AU78Dennis KimagIs9 III
Strain, strain background
(C. elegans)		CB6088Jonathan Hodgkinegl-9(sa307) V hif-1(ia4) V
Strain, strain background
(C. elegans)		JT307Creg Darbyegl-9(sa307) V
Strain, strain background
(C. elegans)		MT20483Dengke Ma/Bob
Horvitz		nIs470 IV
Strain, strain background
(C. elegans)		MT22836this papercyp-36A1(gk824636) I ; egl-9(sa307) V
Strain, strain background
(C. elegans)		MT23218this papernIs682 X
Strain, strain background
(C. elegans)		MT24164this papercyp-36A1(gk824636) I ; agIs9 III
Strain, strain background
(C. elegans)		MT24165this papercyp-36A1(gk824636) I ; agIs9 III ;
egl-9(sa307) V
Strain, strain background
(C. elegans)		MT24166this paperagIs9 III ; egl-9(sa307) V hif-1(ia4) V
Strain, strain background
(C. elegans)		MT24167this paperagIs9 III ; egl-9(sa307) V
Strain, strain background
(C. elegans)		MT24177this paperagIs9 III ; hif-1(ia4) V
Strain, strain background
(C. elegans)		MT24179this papercyp-36A1(n5666) I ; (nIs470) IV
Strain, strain background
(C. elegans)		MT24520this papercyp-36A1(n5666) I ; (nIs470) IV ;
egl-9(n586ts) V ; nIs674
Strain, strain background
(C. elegans)		MT24622this papercyp-36A1(n5666) I ; (nIs470) IV ;
egl-9(n586ts) V
Strain, strain background
(C. elegans)		MT24684this papercyp-36A1(gk824636) I ; agIs9 III ;
nhr-46(n6126) IV ; egl-9(sa307) V ;
nEx2586
Strain, strain background
(C. elegans)		MT24690this paperagIs9 III ; otEx3165
Strain, strain background
(C. elegans)		MT24692this paperagIs9 III ; otEx3156
Strain, strain background
(C. elegans)		MT24911this papercyp-36A1(gk824636) I ; agIs9 III ;
nhr-46(n6126) IV ; egl-9(sa307) V
Strain, strain background
(C. elegans)		MT25047this paperagIs9 III ; nEx2699
Strain, strain background
(C. elegans)		MT25048this papercyp-36A1(gk824636) I ; agIs9 III ;
nEx2699
Strain, strain background
(C. elegans)		MT25050this papercyp-36A1(gk824636) I ; agIs9 III ;
otEx3156
Strain, strain background
(C. elegans)		MT25051this papercyp-36A1(gk824636) I ; agIs9 III ;
otEx3165
Strain, strain background
(C. elegans)		MT25104this papercyp-36A1(gk824636) I ; agIs9 III ;
nhr-46(n6126) IV ; egl-9(sa307) V ;
nEx2713
Strain, strain background
(C. elegans)		MT25107this papercyp-36A1(gk824636) I ; agIs9 III ;
nhr-46(n6126) IV ; egl-9(sa307) V ;
nEx2715
Strain, strain background
(C. elegans)		MT25196this paperagIs9 III ; nhr-46(n6125) IV
Strain, strain background
(C. elegans)		MT25197this papercyp-36A1(gk824636) I ; agIs9 III ;
nhr-46(n6125) IV ; egl-9(sa307) V
Strain, strain background
(C. elegans)		MT25211this papercyp-36A1(gk824636) I
Strain, strain background
(C. elegans)		MT25212this papercyp-36A1(gk824636) I ; egl-9(n586ts) V
Strain, strain background
(C. elegans)		MT25213this papercyp-36A1(gk824636) I ;
nhr-46(n6126) IV ; egl-9(n586ts) V
Strain, strain background
(C. elegans)		MT25214this papernhr-46(n6126) IV
Strain, strain background
(C. elegans)		MT25215this paperegl-9(n586ts) V
Strain, strain background
(C. elegans)		MT25596this papercyp-36A1(gk824636) I ; agIs9 III ;
egl-9(sa307) V ; nEx2853
Strain, strain background
(C. elegans)		MT25599this papercyp-36A1(gk824636) I ; agIs9 III ;
egl-9(sa307) V ; nEx2856
Strain, strain background
(C. elegans)		MT25601this papercyp-36A1(gk824636) I ; agIs9 III ;
egl-9(sa307) V ; nEx2849
Strain, strain background
(C. elegans)		MT25605this papercyp-36A1(gk824636) I ; agIs9 III ;
egl-9(sa307) V ; nEx2859
Strain, strain background
(C. elegans)		MT25606this paperagIs9 III ; nEx2860
Strain, strain background
(C. elegans)		MT25610this papercyp-36A1(gk824636) I ; agIs9 III ;
nhr-46(n6126) IV ; egl-9(sa307) V ; nEx2864
Strain, strain background
(C. elegans)		MT25611this papercyp-36A1(gk824636) I ; agIs9 III ; nEx2860
Strain, strain background
(C. elegans)		MT25627this paperagIs9 III ; nhr-46(n6126) IV
Strain, strain background
(C. elegans)		ZG31Huaqi Jiang/Jo Anne Powell-Coffmanhif-1(ia4) V

Additional files

Supplementary file 1

Genes that were significantly differentially expressed between wild type and egl-9(lf) and at least twofold downregulated in egl-9(lf) vs. wild type.

https://doi.org/10.7554/eLife.36828.019
Supplementary file 2

Genes that were significantly differentially expressed between wild type and egl-9(lf) and at least twofold upregulated in egl-9(lf) vs. wild type.

https://doi.org/10.7554/eLife.36828.020
Transparent reporting form
https://doi.org/10.7554/eLife.36828.021

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