1.Progress in the mechanisms of response to different oxygen concentrations in Caenorhabditis elegans.
Ping HE ; Na KANG ; Ping-Ping LOU ; Hai-Jun TU
Acta Physiologica Sinica 2020;72(3):407-417
Oxygen levels are unequal in different living geographical locations of human and related to normal physiology of health. The reduction of oxygen level in the body can lead to a variety of diseases, such as stroke caused by cerebral ischemia and hypoxia. In the recent years, many studies have elucidated the molecular and cellular mechanisms of organism response to different oxygen concentrations by using the nematode Caenorhabditis elegans (C. elegans) as model organism. C. elegans can escape hypoxia or hyperoxia and adapt to the ambient oxygen environments, and there are different response and regulation mechanisms in different degrees of hypoxia environment. In this paper, recent advances in the reaction of nematodes to different oxygen concentrations and the underlying mechanism were reviewed.
Animals
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Caenorhabditis elegans
;
Caenorhabditis elegans Proteins
;
Humans
;
Hypoxia
;
Oxygen
2.Cold stress reduces lifespan and mobility of C. elegans by mediating lipid metabolism disorder and abnormal stress.
Hao SHI ; Chao ZHANG ; Jia Min ZHAO ; Yi Wen LI ; Yun Jia LI ; Jun Jie LI ; Zhi Yun ZENG ; Lei GAO
Journal of Southern Medical University 2022;42(8):1159-1165
OBJECTIVE:
To investigate the changes of lipid metabolism and stress response of adult C.elegans exposed to non-freezing low temperature and explore the possible mechanism.
METHODS:
The survival rate and activity of adult C.elegans cultured at 20℃ or 4℃ were observed.Lipid metabolism of the cultured adult C.elegans was evaluated using oil red O staining and by detecting the expressions of the genes related with lipid metabolism.The effects of low temperature exposure on stress level of adult C.elegans were evaluated using mitochondrial fluorescence staining and by detecting the expression levels of stress-related genes and antioxidant genes at both the mRNA and protein levels.
RESULTS:
The lifespan and activity of adult C.elegans exposed to low temperature were significantly reduced with decreased lipid accumulation (P < 0.05) and decreased expressions of genes related with fatty acid synthesis and metabolism (fat-5, fat-6, fat-7, fasn-1, nhr-49, acs-2 and aco-1;P < 0.01).Cold stress significantly increased the expressions of heat shock proteins hsp-70 and hsp16.2(P < 0.05) but lowered the number of mitochondria (P < 0.0001) and the expressions of atfs-1, sod-2, sod-3 and gpx-1(P < 0.05).Knockout of fat-5, nhr-49 or both fat-5 and fat-6 obviously enhanced the sensitivity of C.elegans to cold stress as shown by further reduced activity (P < 0.05) and reduced survival rate at 24 h (P < 0.0001) under cold stress.
CONCLUSION
Exposure to a low temperature at 4℃ results in lowered lipid metabolism of adult C.elegans accompanied by a decreased mitochondrial number and quality control ability, which triggers high expressions of stress-related genes and causes reduction of antioxidant capacity, thus callsing lowered activity and reduced lifespan of C.elegans.
Animals
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Antioxidants/metabolism*
;
Caenorhabditis elegans
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Caenorhabditis elegans Proteins/genetics*
;
Cold-Shock Response
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Lipid Metabolism
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Lipid Metabolism Disorders
;
Longevity/genetics*
3.Antioxidant and antiaging effect of traditional Thai rejuvenation medicines in Caenorhabditis elegans.
Sutticha RUANGCHUAY ; Qiang-Qiang WANG ; Liang-Yi WANG ; Jing LIN ; Yong-Chao WANG ; Guo-Huan ZHONG ; Katesarin MANEENOON ; Ze-Bo HUANG ; Sasitorn CHUSRI
Journal of Integrative Medicine 2021;19(4):362-373
OBJECTIVE:
This study explored the rejuvenation mechanisms of Thai polyherbal medicines using different approaches, including in vitro methods, as well as a well-defined nematode model, Caenorhabditis elegans.
METHODS:
THP-R-SR012 decoction was selected from 23 polyherbal medicines, based on metal-chelating and chain-breaking antioxidant capacities. The influences of this extract on the survival and some stress biomarkers of C. elegans under paraquat-induced oxidative stress were evaluated. Furthermore, lifespan analysis and levels of lipofuscin accumulation were examined in senescent nematodes. The phytochemical profile of THP-R-SR012 was analyzed.
RESULTS:
Supplementation with THP-R-SR012 decoction significantly increased the mean lifespan and reduced the oxidative damage to C. elegans under oxidative stress conditions. Further, THP-R-SR012 supplementation slightly influenced the lifespan and the level of lipofuscin accumulation during adulthood. Antioxidant-related phytochemical constituents of THP-R-SR012 decoction were rutin, naringenin, 3,4-dihydroxybenzoic acid, gallic acid, glycyrrhizic acid, demethoxycurcumin and 18α-glycyrrhetinic acid.
CONCLUSION
The antioxidant potential of THP-R-SR012 was due to its scavenging properties, its enhancement of antioxidant-related enzyme activities, and the presence of the antioxidant-related compound. These results support the traditional use of THP-R-SR012 decoction as a tonic for nourishing and strengthening the whole body.
Animals
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Antioxidants/pharmacology*
;
Caenorhabditis elegans/metabolism*
;
Caenorhabditis elegans Proteins/metabolism*
;
Oxidative Stress
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Plant Extracts/pharmacology*
;
Reactive Oxygen Species
;
Rejuvenation
;
Thailand
4.Identifying interacting proteins of a Caenorhabditis elegans voltage-gated chloride channel CLH-1 using GFP-Trap and mass spectrometry.
Zi-Liang ZHOU ; Jing JIANG ; Jiang-An YIN ; Shi-Qing CAI
Acta Physiologica Sinica 2014;66(3):341-348
Chloride channels belong to a superfamily of ion channels that permit passive passage of anions, mainly chloride, across cell membrane. They play a variety of important physiological roles in regulation of cytosolic pH, cell volume homeostasis, organic solute transport, cell migration, cell proliferation, and differentiation. However, little is known about the functional regulation of these channels. In this study, we generated an integrated transgenic worm strain expressing green fluorescence protein (GFP) fused CLC-type chloride channel 1 (CLH-1::GFP), a voltage-gated chloride channel in Caenorhabditis elegans (C. elegans). CLH-1::GFP was expressed in some unidentified head neurons and posterior intestinal cells of C. elegans. Interacting proteins of CLH-1::GFP were purified by GFP-Trap, a novel system for efficient isolation of GFP fusion proteins and their interacting factors. Mass spectrometry (MS) analysis revealed that a total of 27 high probability interacting proteins were co-trapped with CLHp-1::GFP. Biochemical evidence showed that eukaryotic translation elongation factor 1 (EEF-1), one of these co-trapped proteins identified by MS, physically interacted with CLH-1, in consistent with GFP-Trap experiments. Further immunostaining data revealed that the protein level of CLH-1 was significantly increased upon co-expression with EEF-1. These results suggest that the combination of GFP-Trap purification with MS is an excellent tool to identify novel interacting proteins of voltage-gated chloride channels in C. elegans. Our data also show that EEF-1 is a regulator of voltage-gated chloride channel CLH-1.
Animals
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Animals, Genetically Modified
;
Caenorhabditis elegans
;
genetics
;
metabolism
;
Caenorhabditis elegans Proteins
;
metabolism
;
Chloride Channels
;
metabolism
;
Green Fluorescent Proteins
;
chemistry
;
Mass Spectrometry
;
Peptide Elongation Factor 1
;
metabolism
5.Development and application of the physical hypoxic models of C. elegans.
Chang-hong REN ; Ji-ye ZHANG ; Jin-ping SHI ; Bin JIANG ; Na ZHAO ; Hu-qi LIU ; Cheng-gang ZHANG
Chinese Journal of Applied Physiology 2011;27(3):257-262
OBJECTIVETo develop a suitable hypoxic injury model, which is important for revealing pathological molecular mechanism of hypoxia.
METHODSWe focused on C. elegans by treatment with different hypoxic times and systematically observed mortality, movement, Cellular morphology and the related-protein expression of the animals.
RESULTSWe demonstrated that hypoxia (0.2% partial pressure of oxygen) induced morphological cell defects, and then leading to death of C. elegans. The mortality of C. elegans increased along with hypoxic time, while hypoxia-inducible factor (HIF-1) was significantly up-regulated. In addition, by using neuron-specific transgenic wonns with green fluorescent protein--we observed the neuron-specffic injury caused by hypoxic stress.
CONCLUSIONWe successfully established an effective, convenient physical hypoxic model of C. elegans, which will facilitate the studies of hypoxic pathology and molecular mechanisms of hypoxic response in the future.
Animals ; Caenorhabditis elegans ; physiology ; Caenorhabditis elegans Proteins ; metabolism ; Disease Models, Animal ; Hypoxia ; physiopathology ; Hypoxia-Inducible Factor 1 ; metabolism ; Neurons ; pathology ; Transcription Factors ; metabolism
6.Systematic temperature signaling regulates behavior plasticity.
Protein & Cell 2011;2(10):774-775
7.C30F12.4 influences oogenesis, fat metabolism, and lifespan in C. elegans.
Lu WANG ; Fei XU ; Guishuan WANG ; Xiaorong WANG ; Ajuan LIANG ; Hefeng HUANG ; Fei SUN
Protein & Cell 2016;7(10):714-721
Reproduction, fat metabolism, and longevity are intertwined regulatory axes; recent studies in C. elegans have provided evidence that these processes are directly coupled. However, the mechanisms by which they are coupled and the reproductive signals modulating fat metabolism and lifespan are poorly understood. Here, we find that an oogenesis-enriched gene, c30f12.4, is specifically expressed and located in germ cells and early embryos; when the gene is knocked out, oogenesis is disrupted and brood size is decreased. In addition to the reproductive phenotype, we find that the loss of c30f12.4 alters fat metabolism, resulting in decreased fat storage and smaller lipid droplets. Meanwhile, c30f12.4 mutant worms display a shortened lifespan. Our results highlight an important role for c30f12.4 in regulating reproduction, fat homeostasis, and aging in C. elegans, which helps us to better understand the relationship between these processes.
Animals
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Caenorhabditis elegans
;
genetics
;
metabolism
;
Caenorhabditis elegans Proteins
;
genetics
;
metabolism
;
Female
;
Lipid Droplets
;
metabolism
;
Lipid Metabolism
;
physiology
;
Longevity
;
physiology
;
Mutation
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Oogenesis
;
physiology
8.Expression changes of age-related genes in different aging stages of Caenorhabiditis elegans and the regulating effects of Chuanxiong extract.
Xiaoyan WANG ; Xiangming WANG ; Danqiao WANG ; Lianda LI ; Xiaohong NIU
China Journal of Chinese Materia Medica 2010;35(12):1599-1602
OBJECTIVETo explore the expression changes of age-related genes in different stages of aging and the regulating effects of Chuanxiong extract on it.
METHODAccording to the different stages of aging, the experiments were tested at two time points of 2 d and 6 d. Using realtime RT-PCR (qRT-PCR) to test the expression change of aging-related genes among the groups.
RESULTCompared with the 2 d control group, the expression of age-1, daf-2, let-363 were up-regulated in the 6 d control group (P < 0.05) while the expression of ins-18, let-60, sir-2.1, sod-3 were down-regulated (P < 0.05). Compared with the 2 d administration group, the expression of age-1, daf-2, let-363 were significantly up-regulated (P < 0.01) in the 6 d administration group after treated with CXE while the expression of ins-18, let-60, sir-2.1, sod-3 were significantly down-regulated (P < 0.01).
CONCLUSIONIn the progress of aging, the expression of age-1, daf-2, let-363 increased, functioning as aging-promoting genes; while the expression of ins-18, let-60, sir-2.1, sod-3 decreased, functioning as longevity genes; CXE extended the lifespan through inhibiting the expression of these aging-promoting genes and increasing the expression of longevity genes, which would be the molecular mechaniSm of anti-aging of traditional Chinese medicine that can promote Qi and activate blood.
Animals ; Caenorhabditis elegans ; genetics ; growth & development ; metabolism ; Caenorhabditis elegans Proteins ; genetics ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Gene Expression Regulation, Developmental ; drug effects ; Longevity ; drug effects
9.Toxicity and metabolism of 3-bromopyruvate in Caenorhabditis elegans.
Qiao-Ling GU ; Yan ZHANG ; Xi-Mei FU ; Zhao-Lian LU ; Yao YU ; Gen CHEN ; Rong MA ; Wei KOU ; Yong-Mei LAN
Journal of Zhejiang University. Science. B 2020;21(1):77-86
In this study, we aimed to evaluate the toxic effects, changes in life span, and expression of various metabolism-related genes in Caenorhabditis elegans, using RNA interference (RNAi) and mutant strains, after 3-bromopyruvate (3-BrPA) treatment. C. elegans was treated with various concentrations of 3-BrPA on nematode growth medium (NGM) plates, and their survival was monitored every 24 h. The expression of genes related to metabolism was measured by the real-time fluorescent quantitative polymerase chain reaction (qPCR). Nematode survival in the presence of 3-BrPA was also studied after silencing three hexokinase (HK) genes. The average life span of C. elegans cultured on NGM with 3-BrPA was shortened to 5.7 d compared with 7.7 d in the control group. hxk-1, hxk-2, and hxk-3 were overexpressed after the treatment with 3-BrPA. After successfully interfering hxk-1, hxk-2, and hxk-3, the 50% lethal concentration (LC50) of all mutant nematodes decreased with 3-BrPA treatment for 24 h compared with that of the control. All the cyp35 genes tested were overexpressed, except cyp-35B3. The induction of cyp-35A1 expression was most obvious. The LC50 values of the mutant strains cyp-35A1, cyp-35A2, cyp-35A4, cyp-35B3, and cyp-35C1 were lower than that of the control. Thus, the toxicity of 3-BrPA is closely related to its effect on hexokinase metabolism in nematodes, and the cyp-35 family plays a key role in the metabolism of 3-BrPA.
Animals
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Caenorhabditis elegans/metabolism*
;
Caenorhabditis elegans Proteins/genetics*
;
Cytochrome P-450 Enzyme System/genetics*
;
Hexokinase/physiology*
;
Pyruvates/toxicity*
;
RNA, Messenger/analysis*
10.A pair of transporters controls mitochondrial Zn2+ levels to maintain mitochondrial homeostasis.
Tengfei MA ; Liyuan ZHAO ; Jie ZHANG ; Ruofeng TANG ; Xin WANG ; Nan LIU ; Qian ZHANG ; Fengyang WANG ; Meijiao LI ; Qian SHAN ; Yang YANG ; Qiuyuan YIN ; Limei YANG ; Qiwen GAN ; Chonglin YANG
Protein & Cell 2022;13(3):180-202
Zn2+ is required for the activity of many mitochondrial proteins, which regulate mitochondrial dynamics, apoptosis and mitophagy. However, it is not understood how the proper mitochondrial Zn2+ level is achieved to maintain mitochondrial homeostasis. Using Caenorhabditis elegans, we reveal here that a pair of mitochondrion-localized transporters controls the mitochondrial level of Zn2+. We demonstrate that SLC-30A9/ZnT9 is a mitochondrial Zn2+ exporter. Loss of SLC-30A9 leads to mitochondrial Zn2+ accumulation, which damages mitochondria, impairs animal development and shortens the life span. We further identify SLC-25A25/SCaMC-2 as an important regulator of mitochondrial Zn2+ import. Loss of SLC-25A25 suppresses the abnormal mitochondrial Zn2+ accumulation and defective mitochondrial structure and functions caused by loss of SLC-30A9. Moreover, we reveal that the endoplasmic reticulum contains the Zn2+ pool from which mitochondrial Zn2+ is imported. These findings establish the molecular basis for controlling the correct mitochondrial Zn2+ levels for normal mitochondrial structure and functions.
Animals
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Caenorhabditis elegans/metabolism*
;
Cation Transport Proteins/genetics*
;
Homeostasis
;
Mitochondria/metabolism*
;
Zinc/metabolism*