cemm agarose plates  (Technical Manufacturing Company)

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: EAT-2 attenuates C. elegans development via metabolic remodeling in a chemically defined food environment

doi: 10.1007/s00018-023-04849-x

Figure Lengend Snippet: eat-2 is expressed in multiple pharyngeal muscles and attenuates C. elegans development on CeMM diet. A N2, 10 trials, n = 45 worms on average in each trial. eat-2(ad1113), 8 trials, n = 58 worms on average in each trial. eat-2(ad1113); Peat-2::eat-2 DNA transgenic rescue line, 3 trials, n = 48 worms on average in each trial. The value represents the percentage of adulthood per day. Fisher’s exact test, ***P < 0.001. B Schematic diagram of YFP knocking into eat-2 genome in eat-2(rg552). Figure adapted from Ref. [21]. C–E Spinning disk confocal microscope photographs. White arrows indicate expression of eat-2 in pharyngeal muscles

Article Snippet: After washing twice with sterile water, eggs were collected and cultured on unseeded NGM plates at 20 °C for ~ 18 h. RNA-seq and analysis Synchronized L1s of different strains (N2, eat-2(ad1113) mutant, tmc-1(rg1003) mutant or eat-2(ad1113);elo-6(lzq11) double mutant) were transferred to corresponding 9 cm CeMM agarose plates.

Techniques: Muscles, Transgenic Assay, Microscopy, Expressing

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: EAT-2 attenuates C. elegans development via metabolic remodeling in a chemically defined food environment

doi: 10.1007/s00018-023-04849-x

Figure Lengend Snippet: Fatty acid metabolic remodeling regulates C. elegans development on CeMM. A–C Dietary supplementation with C17ISO (A), PA (B) and SA (C) promoted nematodes development on CeMM. Control group with 2.5% DMSO, three trials, n = 60 worms on average in each trial; C17ISO, five trials, n = 57 worms on average in each trial; PA, eight trials, n = 56 worms on average in each trial; SA, six trials, n = 60 worms on average in each trial. D Dietary supplementation with ALA (C18:3 n3) did not affect nematode development on CeMM. Control group with 2.5% DMSO, three trials, n = 52 worms on average in each trial; ALA, three trials, n = 53 worms on average in each trial. E Mutation in C17ISO synthesis gene elo-5 attenuated eat-2 mutant development. eat-2(ad1113), 3 trials, n = 60 worms on average in each trial. eat-2(ad1113);elo-5(lzq27), 3 trials, n = 55 worms on average in each trial. F Mutations in C17ISO synthesis gene elo-6 caused eat-2 mutant developmental delay on CeMM. eat-2(ad1113), 3 trials, n = 60 worms on average in each trial. eat-2(ad1113);elo-6(lzq11), 3 trials, n = 59 worms on average in each trial. eat-2(ad1113);elo-6(lzq12), 7 trials, n = 57 worms on average in each trial. eat-2(ad1113);elo-6(lzq19), nine trials, n = 58 worms on average in each trial. G Transgenic whole worm and tissue specific rescue of the development of eat-2;elo-6 double mutants on CeMM. eat-2(ad1113);elo-6(lzq19);Pelo-6::elo-6 DNA, 3 trials, n = 57 worms on average in each trial. eat-2(ad1113);elo-6(lzq11) double mutant transgenic rescue experiments, 2–4 trials, n = 45–80 worms for each trial. H, I Three trials were performed in all groups. eat-2(ad1113), n = 57 worms on average in each trial. eat-2(ad1113);elo-5(lzq27) + DMSO, n = 35 worms on average in each trial; eat-2(ad1113);elo-5(lzq27) + C17ISO, n = 37 worms on average in each trial. eat-2(ad1113);elo-6(lzq11) + DMSO, n = 59 worms on average in each trial; eat-2(ad1113);elo-6(lzq11) + C17ISO, n = 58 worms on average in each trial. *P < 0.05, **P < 0.01, ***P < 0.001, ns: not significant. P values were determined by Fisher’s exact test for (A–E), (H) and (I), the one-way ANOVA with Dunnett’s correction was applied in (F) and (G)

Article Snippet: After washing twice with sterile water, eggs were collected and cultured on unseeded NGM plates at 20 °C for ~ 18 h. RNA-seq and analysis Synchronized L1s of different strains (N2, eat-2(ad1113) mutant, tmc-1(rg1003) mutant or eat-2(ad1113);elo-6(lzq11) double mutant) were transferred to corresponding 9 cm CeMM agarose plates.

Techniques: Control, Mutagenesis, Transgenic Assay

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: EAT-2 attenuates C. elegans development via metabolic remodeling in a chemically defined food environment

doi: 10.1007/s00018-023-04849-x

Figure Lengend Snippet: The metabolomics signature of eat-2;elo-6 double mutants and wild-type worms with dietary C17ISO supplementation. A Heatmap of metabolites abundance significantly changed in eat-2;elo-6 double mutant and wild-type worms with C17ISO supplementation. Metabolites with significantly higher relative abundance level are represented by color red, while metabolites with significantly lower relative abundance level are represented by color blue. Amino acids and amino acid derivatives are highlighted in green; vitamins are highlighted in purple. Details of the metabolome data are represented in Supplementary file 8 and 9. The corresponding full names of some metabolites abbreviations are: 5N-3C 3-(2-P)H, 5-nitrothiophene-3-carbaldehyde 3-(2-pyridyl)hydrazone; (2S)-4O-2P-3,4D-2H-C-7yl β-d-G, (2S)-4-Oxo-2-phenyl-3,4-dihydro-2H-chromen-7-yl beta-d-glucopyranoside; N-[4-(A)P]-2C-6F, N-[4-(aminosulfonyl)phenyl]-2-chloro-6-fluorobenzamide; N-[3-(4C)-1H-1,2,4T-5yl]-N′-I, N-[3-(4-chloroanilino)-1H-1,2,4-triazol-5-yl]-N′-isopropylurea. B SAM can be synthesized from methionine, which could be converted from methionine sulfoxide. C, D SAM (1.5 mM) and methionine sulfoxide (8 mM) significantly promoted the growth of wild-type worms N2 on CeMM. Control group water, three trials, n = 38 worms on average in each trial; SAM, three trials, n = 30 worms on average in each trial; methionine sulfoxide, three trials, n = 39 worms on average in each trial. P values were determined by Fisher’s exact test, *P < 0.05, **P < 0.01, ***P < 0.001

Article Snippet: After washing twice with sterile water, eggs were collected and cultured on unseeded NGM plates at 20 °C for ~ 18 h. RNA-seq and analysis Synchronized L1s of different strains (N2, eat-2(ad1113) mutant, tmc-1(rg1003) mutant or eat-2(ad1113);elo-6(lzq11) double mutant) were transferred to corresponding 9 cm CeMM agarose plates.

Techniques: Mutagenesis, Synthesized, Control

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: EAT-2 attenuates C. elegans development via metabolic remodeling in a chemically defined food environment

doi: 10.1007/s00018-023-04849-x

Figure Lengend Snippet: Proposed model for C. elegans developmental regulation via fatty acid metabolic remodeling on CeMM. The dotted line with arrows on both sides indicates correlation. ACh acetylcholine; SAM S-adenosylmethionine. In wild-type C. elegans, the acetylcholine receptor EAT-2 in the pharyngeal muscle is responsible for receiving upstream acetylcholine neural signals from TMC-1 in the CeMM food environment, and might attenuate animal development by inhibiting fatty acids synthesis and promoting fatty acids β-oxidation. In the absence of this negative regulation, expression of elo-2, elo-5 and elo-6 is up-regulated in eat-2 and tmc-1 mutants, resulting in increased levels of related fatty acids PA, SA and C17ISO, which promotes the development of C. elegans. The development promoting effect of elo-6/C17ISO may be correlated to the up-regulated expression of cuticle synthesis and hedgehog signaling genes, as well as increased biosynthesis of amino acids, amino acid derivatives and vitamins. In particular, the amino acid derivative SAM can significantly promote the C. elegans development

Article Snippet: After washing twice with sterile water, eggs were collected and cultured on unseeded NGM plates at 20 °C for ~ 18 h. RNA-seq and analysis Synchronized L1s of different strains (N2, eat-2(ad1113) mutant, tmc-1(rg1003) mutant or eat-2(ad1113);elo-6(lzq11) double mutant) were transferred to corresponding 9 cm CeMM agarose plates.

Techniques: Expressing