Luqin-like RYamide peptides regulate food-evoked responses in C. elegans
- Graduate School of Science, The University of Tokyo, Japan
- National Cerebral and Cardiovascular Center Research Institute, Japan
- Institute of Life Sciences, Kurume University, Japan
- Frontier Science Research Center, University of Miyazaki, Japan
- University of Miyazaki, Japan
Figures
Figure 1 with 2 supplements
Purification of LURY-1 peptides.
(A) Gel filtration on Sephadex G-50 (fine) of the SP-III fraction obtained from C. elegans. Red bars indicate fluorescence changes owing to [Ca2+]i increase in receptor expressing cells. (top) …
Figure 1—figure supplement 1
Phylogenetic analysis of C. elegans NPR-22 with neuropeptide GPCRs.
Phylogenetic tree constructed by Maximum likelihood analysis using the ‘One Click’ mode (without Gblocks) on the online Phylogeny.fr platform (http://www.phylogeny.fr/, Dereeper et al., 2008). The …
Figure 1—figure supplement 2
Identification of LURY-1 peptides.
(A, B) Final purification of the active peptides P1 (A) and P2 (B) from fraction A on CM-HPLC at pH 6.5. (C) Chromatographic comparison using RP-HPLC of natural LURY-1 peptides and synthetic LURY-1 …
Figure 2 with 1 supplement
LURY-1 peptides are potent ligands for NPR-22.
(A) Genomic organization of the lury-1/Y75B8A.11 (left) and npr-22 (right) loci and the lesions in mutant alleles. (B) Amino acid sequences of LURY-1-1 (red) and LURY-1-2 (blue) in the LURY-1 …
Figure 2—figure supplement 1
Pharmacological characterization of synthetic peptides.
(A, B) Responses to synthetic peptides in CHO cells transiently expressing Drosophila CG5811. Dose-response relationships of changes in [Ca2+]i for (A) dRYamide-1, dRYamide-2 and (B) LURY-1-1, …
Figure 3 with 2 supplements
LURY-1 peptides are expressed in the pharyngeal M1 and M2 neurons, while NPR-22 is broadly expressed.
(A) Expression pattern of lury-1prom::Venus reporter in the head region of an adult worm. Fluorescence of lury-1prom::venus was specifically observed in the M1 and M2 pharyngeal neurons in adult …
Figure 3—figure supplement 1
Expression patterns of lury-1.
(A) Expression pattern of lury-1prom::Venus reporter in an adult worm. lury-1prom::venus was specifically expressed in pharynx. (B) Co-expression of lury-1prom::mCherry and glr-2prom::Venus in M1. (C…
Figure 3—figure supplement 2
Expression patterns of npr-22.
(A) Expression pattern of npr-22prom::Venus reporter in an adult worm. White arrowheads, ventral nerve cord neurons. (B) Co-expression of npr-22prom::mCherry and flp-15prom::Venus in I2. (C) …
Figure 4 with 2 supplements
Multicopy expression of lury-1(+) causes multiple food-related phenotypes in an NPR-22-dependent manner.
(A) Lateral views near the uterus of synchronized adult worms (36 hr after the L4 white-crescent stage) of wild type, Ex[lury-1(+)], and npr-22(ok1598); Ex[lury-1(+)]. Asterisk, ~30 cell stage eggs …
Figure 4—figure supplement 1
Egg-laying phenotypes of canonical Egl-c mutants.
(A) Number of unlaid eggs in the uterus of adult goa-1(n1134), egl-30(js176gf), eat-16(ce71), and egl-19(n2368gf) worms (36 hr after the L4 white-crescent stage). N = 12 animals. ***p<0.001 (ANOVA …
Figure 4—figure supplement 2
Multicopy expression of LURY-1 extends lifespan in an NPR-22-depndent manner.
(A, B) Lifespans of peIs2413 (A) and peIs2414 (B) strains, in which the lury-1(+) transgene is integrated into genomic DNA. The results of wild type and npr-22(ok1598) are the same as shown in Figure…
Figure 5 with 1 supplement
LURY-1 peptides stimulate egg-laying and suppress feeding through NPR-22.
(A, B) Effects of administration of LURY-1 peptides on the number of retained eggs (A) and pharyngeal pumping (B). Each of synthetic LURY-1 peptides was injected to wild-type and npr-22(-) worms at …
Figure 5—figure supplement 1
Administration of FLP-7–3 does not affect egg-laying or feeding.
(A) Number of unlaid eggs in the uterus of animals carrying an extrachromosomal flp-7(+) array. N = 18 animals. (B) Pharyngeal pumping rate of animals carrying an extrachromosomal flp-7(+) array. N =…
Figure 6 with 1 supplement
LURY-1 peptides are secreted in a food-dependent manner and essential for satiety-induced behavioral changes.
(A) The fluorescence of lury-1prom::lury-1::venus reporter in worms that had been fed (top), fasted (middle), or fasted and refed (bottom). White arrowheads, coelomocytes (CC). Scale bars, 100 μm. (B…
Figure 6—figure supplement 1
The activities of coelomocytes and the lury-1 promoter are unaffected by starvation or refeeding.
(A) Coelomocyte uptake of signal sequence-tagged GFP (ssGFP), which was expressed in body wall muscles under the myo-3 promoter. White arrowheads, coelomocytes. Scale bar, 100 μm. (B) The …
Figure 7 with 1 supplement
NPR-22 acts in MC to control feeding and lifespan.
(A) Pharyngeal pumping rate of animals expressing npr-22a under indicated promoters. N = 13, 12, 12, 12, 12, 12, 12, 12, 12, and 12 animals (from left to right in order of each group). (B) Schematic …
Figure 7—figure supplement 1
Both NPR-22a and NPR-22b are functional for the regulation of feeding.
(A) Pharyngeal pumping rate of animals expressing the npr-22a or npr-22b isoform. N = 12 animals. (B) Pharyngeal pumping rate of animals expressing npr-22a under the acr-2 promoter. N = 16 animals. …
Figure 8 with 1 supplement
NPR-22 acts in RIH to control egg-laying.
(A) Number of unlaid eggs in the uterus of animals expressing npr-22a under indicated promoters. N = 18, 18, 18, 19, 18, 18, 20, 18, 18, 19, and 18 animals (from left to right in order of each …
Figure 8—figure supplement 1
Both NPR-22a and NPR-22b are functional for the regulation of egg-laying.
(A) Number of unlaid eggs in the uterus of animals expressing the npr-22a or npr-22b isoform. N = 18, 18, 18, 18, 16, and 18 animals (from left to right in order of each group). (B) Number of unlaid …
Figure 9
Serotonin signaling is essential for the LURY-1 peptide-dependent stimulation of egg-laying.
(A) Genetic interaction between peIs2413[lury-1(+)] and monoamine-related genes in number of unlaid eggs in the uterus. N = 20 animals. (B) Changes in number of unlaid eggs in the uterus. Animals …
Tables
Table 1
Active fractions of each chromatography and the amino acid sequences of the purified peptides.
https://doi.org/10.7554/eLife.28877.007| Ion-exchange- HPLC Elution time (min) | Reversed-phase- HPLC Retention time (min) | Observed mass (MH+) | Theoretical mass (MH+) | Sequence | Precursor gene | Peptide name* | |
|---|---|---|---|---|---|---|---|
| pH 6.5 | pH4.8 | ||||||
| 48–52 (Fr. A) | 46–48 | 13–13.5 | 717.25 | 717.42 | AVLPRY-NH2 | lury-1 | LURY-1-2 |
| 17–18 | 818.34 | 818.46 | PALLSRY-NH2 | lury-1 | LURY-1-1 | ||
| 60–62 (Fr. B) | 56–60 | 25.5–26.5 | 792.29 | 792.43 | PNFLRF-NH2 | flp-1 | FLP-1-6 |
| 92–94 (Fr. C) | 108–112 | 27.5–28.5 | 1255.45 | 1255.68 | RNKFEFIRF-NH2 | flp-12 | FLP-12 |
| 94–96 (Fr. D) | 86–88 | 15.5–16.5 | 901.27 | 901.45 | KSAYMRF-NH2 | flp-6 | FLP-6-1 |
| 90–92 | 23–23.5 | 1108.33 | 1108.55 | SPSAKWMRF-NH2 | flp-22 | FLP-22 | |
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*Peptide names other than LURY-1 peptides are taken from Li and Kim, 2008.
Additional files
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Supplementary file 1
Results of lifespan analyses.
- https://doi.org/10.7554/eLife.28877.023
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Supplementary file 2
Strains used in this study.
- https://doi.org/10.7554/eLife.28877.024
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Transparent reporting form
- https://doi.org/10.7554/eLife.28877.025
