Wild-type N2 Research Articles

LIR-1 and PDTF-1 regulate the permeability barrier function of the C. elegans cuticle.

The cuticle of C. elegans presents the second largest surface area in the soma of worms. We have recently reported that the permeability barrier in the cuticle is dependent on six permeability-determining (PD) collagens and BLMP-1, a transcription factor. To identify additional regulators of cuticle permeability, we performed RNA interference for 286 transcription factors expressed in the C. elegans hypodermis and studied cuticle permeability to Hoechst 33258, a nucleic acid dye. Although the cuticle of wild type N2 strain is impermeable to this dye, RNAi of lir-1 or T26A8.1 (PDTF-1) caused permeability defect in the cuticle. LIR- 1 and PDTF-1 positively regulate expression of collagen and/or collagen processing enzymes. As a consequence, lir-1 and pdtf-1 RNAi also caused enhanced susceptibility to exogenous toxins such as paraquat, levamisole and ivermectin. Thus, LIR-1 and PDTF-1 are two hypodermis-specific transcriptional regulators of cuticle permeability.

Deficient mechanosensation in mec-3 decreases precipice response in C. elegans.

The precipice response in Caenorhabditis elegans is a little-understood phenomenon in which worms move rapidly away from edges. We hypothesized that mechanosensation underlies the precipice response and that mechanosensory mutants would exhibit the precipice response less often than N2 wild type worms. We found that mec-3 mutants, with severe loss of mechanosensation, exhibited the precipice response at a lower rate than N2, but mec-10 and trp-4 mutants, with partial loss of response to mechanical stimuli, responded at a similar rate to N2. These results provide a characterization of the precipice response and implicate a role for mechanosensation in this behavior.

Paraquat exposure over generation affects lifespan and reproduction through mitochondrial disruption in C. elegans

Paraquat (methyl viologen), is a non-selective contact herbicide and well known mitochondrial toxicant. Mitochondria are the center of cellular metabolism, and involved in the development, lifespan, and reproduction of an organism. Mitochondria are dynamic organelles that are inherited maternally through the germline and carry multiple copies of their own genome (mtDNA). It is important to understand the effects of acute and chronic stress caused by mitochondrial toxicants over multiple generations at the cellular and organism levels. Using the model nematode C. elegans, we show that acute and chronic exposure to paraquat affects reproduction, longevity, gene expression, and mitochondrial physiology. Acute exposure to paraquat in N2 (wild type) causes induction of mitochondrial unfolded protein response (mtUPR), increased expression of mitochondrial superoxide dismutase, decreased mitochondrial membrane potential (Δψm), a dose-dependent progression from linear to fragmented mitochondria, and dose-dependent changes in longevity. Chronic exposure to a low dose of paraquat (0.035 mM) over multiple generations in N2 causes a progressive decline of fertility, leading to complete loss of fertile embryo production by the third generation. The mutation in CEP-1 [cep-1(gk138)], a key regulator of stress-induced apoptosis in the germline, causes increased sensitivity to chronic paraquat relative to N2 with no fertile embryo production beyond the second generation. Whereas, mitochondrial electron transport chain (complex III) mutant [isp-1(qm150)], which display constitutive activation of mtUPR showed increased tolerance and produced fertile embryo out to the fourth generation. The N2, cep-1(gk138), and isp-1(qm150) strain’s lifespan over multiple generations exposed to chronic paraquat were measured. Fertility and lifespan data together indicate a trade-off between reproduction and somatic maintenance during chronic paraquat exposure. We have proposed that mitochondrial signaling, dynamics, and CEP-1 mediated germline apoptosis is involved in this trade-off.

G protein-coupled receptor kinase 2 modifies the ability of Caenorhabditis elegans to survive oxidative stress.

Survival and adaptation to oxidative stress is important for many organisms, and these occur through the activation of many different signaling pathways. In this report, we showed that Caenorhabditis (C.) elegans G protein-coupled receptor kinases modified the ability of the organism to resist oxidative stress. In acute oxidative stress studies using juglone, loss-of-function grk-2 mutants were more resistant to oxidative stress compared with loss-of-function grk-1 mutants and the wild-type N2 animals. This effect was Ce-AKT-1 dependent, suggesting that Ce-GRK2 adjusted C. elegans oxidative stress resistance through the IGF/insulin-like signaling (IIS) pathway. Treating C. elegans with a GRK2 inhibitor, the selective serotonin reuptake inhibitor paroxetine, resulted in increased acute oxidative stress resistance compared with another selective serotonin reuptake inhibitor, fluoxetine. In chronic oxidative stress studies with paraquat, both grk-1 and grk-2 mutants had longer lifespan compared with the wild-type N2 animals in stress. In summary, this research showed the importance of both GRKs, especially GRK2, in modifying oxidative stress resistance.

Pepper component 7-ethoxy-4-methylcoumarin, a novel dopamine D2 receptor agonist, ameliorates experimental Parkinson’s disease in mice and Caenorhabditis elegans

Parkinson’s disease (PD) is a progressive neurodegenerative disease resulting from the degeneration of dopaminergic (DAergic) neurons in the substantia nigra pars compacta (SNpc) and subsequent deficit of dopamine in the striatum. PD is inversely associated with consumption of peppers; however, the constituent and the underlying mechanism remain unclear. This study aimed to investigate the effects of 7-ethoxy-4-methylcoumarin (EMC), a pepper constituent, on PD-like disorders in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced mice and 6-hydroxydopamine (6-OHDA)-exposed C. elegans. In this study, EMC was identified as an agonist of dopamine D2 receptor (DRD2) and increased the expression of P-CREB and BDNF in SH-SY5Y cells. In MPTP-treated PD mice, EMC was shown to apparently ameliorate the motor and gait disorders, and restore the depressed TH expression in SNpc and striatum. Meanwhile, it recovered the locomotor deficit caused by 6-OHDA in wild type N2 and CAT-2-transgenic UA57 of C. elegans, and relieved the degeneration of DAergic neurons resulting from 6-OHDA or with ageing. Moreover, EMC inhibited α-synuclein accumulation in C. elegans strain NL5901 overexpressing human α-synuclein gene. Taken together, EMC was identified as a novel DRD2 agonist and improved experimental PD in mice and C. elegans. These findings suggest that EMC may be beneficial to PD patients, further supporting that the consumption of peppers may have favorable effect on PD progression.

G protein coupled receptor kinases modulate Caenorhabditis elegans reactions to heat stresses

In this report, we explored if G protein coupled receptor kinases (GRKs) can help modulate the heat stress responses of Caenorhabditis (C.) elegans. Loss of function grk-2 C. elegans mutants were more tolerant to increases in heat and display an ability for heat stress-associated hormesis at a longer exposure time unlike the wild type N2 animals and the loss of function grk-1 C. elegans mutants. The loss of function grk-1 mutants recovered more from acute heat stress compared to the wild type N2 animals. Animals with low Ce-GRK2 protein expression showed increased DAF-16 nuclear localization during the early stages of heat stress exposure compared to the other RNAi-treated animals, demonstrating altered insulin/insulin-like growth factor signaling (IIS) pathway activity in response to the stress. pdk-1 and akt-1 may play key roles in conjunction with Ce-GRK2 in the heat stress response. Collectively, these findings demonstrate that GRKs influence C. elegans heat stress behaviors.

Genetic Variation in Complex Traits in Transgenic α-Synuclein Strains of Caenorhabditis elegans.

Different genetic backgrounds can modify the effect of mutated genes. Human α-synuclein (SNCA) gene encodes α-synuclein, and its oligomeric complexes accumulate with age and mediate the disruption of cellular homeostasis, resulting in the neuronal death that is characteristic of Parkinson’s Disease. Polymorphic variants modulate this complex pathologic mechanism. Previously, we constructed five transgenic introgression lines of a Caenorhabditis elegans model of α-synuclein using genetic backgrounds that are genetically diverse from the canonical wild-type Bristol N2. A gene expression analysis revealed that the α-synuclein transgene differentially affects genome-wide transcription due to background modifiers. To further investigate how complex traits are affected in these transgenic lines, we measured the α-synuclein transgene expression, the overall accumulation of the fusion protein of α-synuclein and yellow fluorescent protein (YFP), the lysosome-related organelles, and the body size. By using quantitative PCR (qPCR), we demonstrated stable and similar expression levels of the α-synuclein transgene in different genetic backgrounds. Strikingly, we observed that the levels of the a-synuclein:YFP fusion protein vary in different genetic backgrounds by using the COPAS™ biosorter. The quantification of the Nile Red staining assay demonstrates that α-synuclein also affects lysosome-related organelles and body size. Our results show that the same α-synuclein introgression in different C. elegans backgrounds can produces differing effects on complex traits due to background modifiers.

NHJ-1 Is Required for Canonical Nonhomologous End Joining in Caenorhabditis elegans.

DNA double-strand breaks (DSBs) are a particularly lethal form of DNA damage that must be repaired to restore genomic integrity. Canonical nonhomologous end joining (NHEJ), is a widely conserved pathway that detects and directly ligates the broken ends to repair the DSB. These events globally require the two proteins that form the Ku ring complex, Ku70 and Ku80, and the terminal ligase LIG4. While the NHEJ pathway in vertebrates is elaborated by more than a dozen factors of varying conservation and is similarly complex in other eukaryotes, the entire known NHEJ toolkit in Caenorhabditis elegans consists only of the core components CKU-70, CKU-80, and LIG-4 Here, we report the discovery of the first accessory NHEJ factor in C. elegans Our analysis of the DNA damage response in young larvae revealed that the canonical wild-type N2 strain consisted of two lines that exhibited a differential phenotypic response to ionizing radiation (IR). Following the mapping of the causative locus to a candidate on chromosome V and clustered regularly interspaced short palindromic repeats-Cas9 mutagenesis, we show that disruption of the nhj-1 sequence induces IR sensitivity in the N2 line that previously exhibited IR resistance. Using genetic and cytological analyses, we demonstrate that nhj-1 functions in the NHEJ pathway to repair DSBs. Double mutants of nhj-1 and lig-4 or cku-80 do not exhibit additive IR sensitivity, and the post-IR somatic and fertility phenotypes of nhj-1 mimic those of the other NHEJ factors. Furthermore, in com-1 mutants that permit repair of meiotic DSBs by NHEJ instead of restricting their repair to the homologous recombination pathway, loss of nhj-1 mimics the consequences of loss of lig-4 Diakinesis-stage nuclei in nhj-1; com-1 and nhj-1; lig-4 mutant germlines exhibit increased numbers of DAPI-staining bodies, consistent with increased chromosome fragmentation in the absence of NHEJ-mediated meiotic DSB repair. Finally, we show that NHJ-1 and LIG-4 localize to somatic nuclei in larvae, but are excluded from the germline progenitor cells, consistent with NHEJ being the dominant DNA repair pathway in the soma. nhj-1 shares no sequence homology with other known eukaryotic NHEJ factors and is taxonomically restricted to the Rhabditid family, underscoring the evolutionary plasticity of even highly conserved pathways.

Assessment of the In Vivo Antioxidant Activity of an Anthocyanin-Rich Bilberry Extract Using the Caenorhabditis elegans Model.

Anthocyanins have been associated with several health benefits, although the responsible mechanisms are not well established yet. In the present study, an anthocyanin-rich extract from bilberry (Vaccinium myrtillus L.) was tested in order to evaluate its capacity to modulate reactive oxygen species (ROS) production and resistance to thermally induced oxidative stress, using the nematode Caenorhabditis elegans as an in vivo model. The assays were carried out with the wild-type N2 strain and the mutant strains daf-16(mu86) I and hsf-1(sy441), which were grown in the presence of two anthocyanin extract concentrations (5 and 10 μg/mL in the culture medium) and further subjected to thermal stress. The treatment with the anthocyanin extract at 5 μg/mL showed protective effects on the accumulation of ROS and increased thermal resistance in C. elegans, both in stressed and non-stressed young and aged worms. However, detrimental effects were observed in nematodes treated with 10 μg/mL, leading to a higher worm mortality rate compared to controls, which was interpreted as a hormetic response. These findings suggested that the effects of the bilberry extract on C. elegans might not rely on its direct antioxidant capacity, but other mechanisms could also be involved. Additional assays were performed in two mutant strains with loss-of-function for DAF-16 (abnormal DAuer Formation factor 16) and HSF-1 (Heat Shock Factor 1) transcription factors, which act downstream of the insulin/insulin like growth factor-1 (IGF-1) signaling pathway. The results indicated that the modulation of these factors could be behind the improvement in the resistance against thermal stress produced by bilberry anthocyanins in young individuals, whereas they do not totally explain the effects produced in worms in the post-reproductive development stage. Further experiments are needed to continue uncovering the mechanisms behind the biological effects of anthocyanins in living organisms, as well as to establish whether they fall within the hormesis concept.

Infertility induced by auxin in PX627 Caenorhabditis elegans does not affect mitochondrial functions and aging parameters.

Caenorhabditis elegans is widely used for aging studies. 5-Fluoro-2´-deoxyuridine (FUdR) is commonly used to control offspring. While larvae are stopped from further development, also mitochondrial DNA and function may be affected. Since mitochondria and longevity are closely related, the use of FUdR may falsify possible studies. PX627, an auxin inducible infertility strain to control offspring, allows mitochondrial investigations during senescence without FUdR toxicity.Longevity and health parameters were assessed in 2- and 10-day old nematodes wild-type N2 and PX627 treated with FUdR or auxin, respectively. Mitochondrial membrane potential, energetic metabolites and reactive oxygen species levels, were determined. mRNA expression levels of key genes involved were quantified using quantitative real-time PCR.FUdR significantly increased lifespan and health parameters, as well as, mitochondrial function compared to untreated controls and auxin treated PX627. Although a decrease in all parameters could be observed in aged nematodes, this was less severe after FUdR exposure. Glycolysis was significantly up-regulated in aged PX627 compared to N2. Expression levels of daf-16, sir-2.1, aak-2, skn-1, atp-2 and atfs-1 were regulated accordingly.Hence, auxin in PX627 might be a good alternative to control progeny, for mitochondrial- and longevity-related investigations in nematodes.

Tart Cherry Increases Lifespan in Caenorhabditis elegans by Altering Metabolic Signaling Pathways.

Aging and healthspan are determined by both environmental and genetic factors. The insulin/insulin-like growth factor-1(IGF-1) pathway is a key mediator of aging in Caenorhabditis elegans and mammals. Specifically, DAF-2 signaling, an ortholog of human IGF, controls DAF-16/FOXO transcription factor, a master regulator of metabolism and longevity. Moreover, mitochondrial dysfunction and oxidative stress are both linked to aging. We propose that daily supplementation of tart cherry extract (TCE), rich in anthocyanins with antioxidant properties may exert dual benefits for mitochondrial function and oxidative stress, resulting in beneficial effects on aging in C. elegans. We found that TCE supplementation at 6 μg or 12 μg/mL, increased (p < 0.05) the mean lifespan of wild type N2 worms, respectively, when compared to untreated control worms. Consistent with these findings, TCE upregulated (p < 0.05) expression of longevity-related genes such as daf-16 and aak-2 (but not daf-2 or akt-1 genes) and genes related to oxidative stress such as sod-2. Further, we showed that TCE supplementation increased spare respiration in N2 worms. However, TCE did not change the mean lifespan of daf-16 and aak-2 mutant worms. In conclusion, our findings indicate that TCE confers healthspan benefits in C. elegans through enhanced mitochondrial function and reduced oxidative stress, mainly via the DAF-16 pathway.

A Hot Water Extract of Sideritis scardica Prolongs Life Span and Enhances Heat Shock Resistance in Caenorhabditis elegans

Sideritis scardica is a Lamiaceae plant that is endemic to the alpine zone of the Balkan Peninsula. The tea of S. scardica has been handed down as a “tea of longevity” in the Rhodope region of Bulgaria for an unknown amount of time. In this study, we prepared a hot water extract of S. scardica (SHWE) and examined its effects on both life span and stress response in living tissue using Caenorhabditis elegans and its transgenic mutants. The life span of wild-type N2 worms was prolonged by approximately 15% at the SHWE concentration of 5 µg/mL and approximately 22% at the SHWE concentration of 50 µg/mL, as compared with the control group. The effect of SHWE on the expression of heat shock protein 16.2 (HSP-16.2) under heat stress was investigated using TJ375 worms, a transgenic mutant of C. elegans. In the TJ375 worms pretreated with SHWE, the fluorescence intensity of green fluorescent protein fluorescence, which indicates the expression of HSP-16.2, was significantly increased. In the assay using TJ356 worms, the worms pretreated with SHWE did not show the translocation of DAF-16, a forkhead transcription factor class O homolog, from the cytoplasm to nucleus under heat stress. Additionally, under heat stress, the pretreatment of SHWE improved the survival rate of GR1307 worms, a knockout mutant of daf-16. These results indicate that SHWE enhances HSP-16.2 expression through a stress-response pathway (eg, HSF-1 pathway) other than the DAF-16 pathway, resulting in a prolonged life span of C. elegans under heat stress.

Anti-aging effects on Caenorhabditis elegans of a polysaccharide, O-acetyl glucomannan, from roots of Lilium davidii var. unicolor Cotton

The anti-aging activities on Caenorhabditis elegans of a polysaccharide, O-acetyl glucomannan (LPR), purified from roots of Lilium davidii var. unicolor Cotton, were assessed by observing the mean lifespan, reproduction, pharyngeal pumping and stress response on nematodes. Additionally, the fluorescence intensity of lipofuscin and the level of reactive oxygen species (ROS) were detected. Also the activities of superoxide dismutase (SOD), catalase (CAT) and contents of malondialdehyde (MDA) were determined by the kit method. The results showed that LPR effectively delayed the aging of C. elegans in a dose-dependent manner. When the concentration reached 4 mg/mL, LPR extended the mean lifespan of C. elegans by up to 40%, 61% (P < 0.01) and 50% (P < 0.05) under normal, thermal and oxidative stress culture conditions, respectively. Moreover, LPR remarkably increased the reproduction duration of the nematodes at a concentration of 1 mg/L, and significantly decreased the ROS and lipofuscin level of C. elegans in three dosage groups. Further study illustrated that LPR at 4 mg/mL strongly increased the activity of SOD and CAT by 39.03% (P < 0.01) and 41.89% (P < 0.05), and decreased the lipid peroxidation of MDA level in C. elegans by 52.59% (P < 0.005) compared to a control. It was inferred that LPR provided stress resistance to heat and oxidation, and prolonged the lifespan of wild type N2 C. elegans mainly by elevating the function of nematode antioxidant defense systems and by scavenging free radicals. These findings provided evidence for the anti-aging properties of this polysaccharide from L. davidii.

Design, synthesis, and biological evaluation of 3-amino-2-oxazolidinone derivatives as potent quorum-sensing inhibitors of Pseudomonas aeruginosa PAO1

Due to the increasing resistance of Pseudomonas aeruginosa to most clinically relevant antimicrobials, it is challenging to treat bacterial infection with traditional antibiotics. Quorum sensing can regulate the production of biofilms and virulence factors which are closely related to bacterial resistance. Previously we synthesized a series of oxazolidinone compounds targeting the quorum-sensing transcriptional regulatory protein CviR and ZS-12 showed good activity against Chromobacterium violaceum CV026 quorum-sensing. In this study, eighteen 3-amino-2-oxazolidinone compounds were designed and synthesized using ZS-12 as the lead compound. We initially evaluated the inhibitory activities of novel oxazolidinone compounds against QS using C. violaceum CV026 as a reporter strain. Thirteen compounds showed good activities (IC50 range 3.69–63.58 μM) and YXL-13 inhibition was the most significant (IC50 = 3.686 ± 0.5790 μM) against biofilm formation and virulence factors determination of P. aeruginosa PAO1. In vitro, YXL-13 significantly inhibited the formation of PAO1 biofilm (range 42.98%–17.67%), the production of virulence factors (pyocyanin, elastase, rhamnolipid, and protease), and bacterial motility. Moreover, the combination of YXL-13 with an antibiotic (meropenem trihydrate) could significantly improve the antibiotic susceptibility of biofilm P. aeruginosa PAO1 cells. In vivo, YXL-13 significantly prolonged the lifespan of wildtype Caenorhabditis elegans N2 infected by P. aeruginosa PAO1. In conclusion, YXL-13 is a candidate agent for antibiotic-resistant P. aeruginosa PAO1and provides a method for finding new antibacterial drugs.

Assessment of lead toxicity on locomotion and growth in a nematode Caenorhabditis elegans

Due to anthropogenic activities and natural abundance, lead exposure is a common phenomenon. Neurotoxic and genotoxic effects of lead are widely known. Recent studies have suggested that lead exposure can affect young generation and transfer to the progeny thus posing a great threat for future generation. The present study was focused on lead toxicity in terms of locomotion and growth of Caenorhabditis elegans (N2 wild type) at three sub-lethal doses (3µM, 15 µM and 30 µM) of Pb (NO3)2 for 24 hours (sub-chronic exposure). Caenorhabditis elegans is a nematode with an established eco- toxicity marker model organism, due to its short life cycle and ease to monitor. After lead exposure, significant toxic manifestations were observed in locomotion of the nematode in terms of omega bends (+350% for 30 µM exposure dose, p&lt;0.001), reversals (-26.98%, -49% and -66.35% for 3 µM, 15 µM and 30 µM exposure doses respectively, p&lt;0.001), turn counts (-38.66%, -62.61% and -81.93% for 3 µM, 15 µM and 30 µM exposure doses respectively, p&lt;0.001 ) and peristaltic speed alterations (+97.83%, +225.92% and +454.63% for 3 µM, 15 µM and 30 µM exposure doses respectively, p&lt;0.001). Successive reduction in the body length at lower doses shows remarkable toxic alterations in nematodes. The obtained data may be useful to extrapolate the effects of lead exposure on humans, as many of the similar pathways and cellular processes affected by Pb in humans are also present in C. elegans.

A model Organism for Antihelminthic Activity: Caenorhabditis elegans and Nigella sativa

In our study, antihelminthic activity of Nigella sativa was tested on a model organism Caenorhabditis elegans (C. elegans). N2 (wild type) C. elegans and Escherichia coli OP50 were purchased from the University of Minnesota, Ceanorhabditis Genetic Center for the study. C. elegans were grown in NGM (Nematode Growth Medium) solid culture medium. After synchronization, nematodes in adult form were exposed to Nigella sativa seed oil at concentrations of 1%, 0.1%, 0.01%, 0.001% and 0.0001%. The study continued for 18 days until all nematodes in the experimental and control groups died. Alive and dead nematodes were recorded every day. The nematodes in which pharengeal pumping stopped was recorded dead. According to our findings, all nematodes died at the end of the fourth day at a concentration of 1%. In addition, no alive nematod was observed at the end of the fifth day at concentration of 0.1%; whereas at the end of the 13th day all nematodes died at concentration of 0.01%. When the data were analyzed statistically, the difference between at concentrations of 0.1%, 0.01%, 0.001% and the control group was significant (p<0.05). The fact that helminths are usually macroscopic in size and most of them cannot be produced in the laboratory by culturing is a major obstacle for scientific studies. Our study has shown that C. elegans can be a good model in studies of antihelmintic activity and that Nigella sativa has an anthelmintic effect.

Tetrastigma hemsleyanum leaves extract against acrylamide-induced toxicity in HepG2 cells and Caenorhabditis elegans

Acrylamide (ACR), as a raw material of polyacrylamide that used in water purification, was verified to possess various toxicity. Tetrastigma hemsleyanum (TH) is a medicinal plant widely used to anti-inflammation and anti-tumor in Chinese folks. However, more researches focused on the biological activities in tubers and the leaves were ignored. Thus, the protective effect of Tetrastigma hemsleyanum leaves extract (THLE) against ACR-induced toxicity in HepG2 cells and Caenorhabditis elegans (C. elegans) was explored in this study. In vitro, we observed that THLE attenuated ACR-induced toxicity in HepG2 cell via regulating Akt/mTOR/FOXO1/MAPK signaling pathway. Further research proved that 5-caffeoylquinic acid (5-CA) plays a major role in THLE’s amelioration effect of ACR toxicity. In vivo, it was found that THLE possesses the same protective effect in ACR-treated wild-type N2 C. elegans and daf-2 (-) (deficit in DAF-2) mutants. However, the anti-ACR toxicity effect of THLE in daf-16 (-) mutants (deficit in DAF-16 that homologous to FOXO family in human) was weakened. Our results indicated that THLE exhibited protective effects against ACR-induced toxicity both in HepG2 cells and C. elegans, while DAF-16/FOXO gene is involved in THLE’ protective effect via regulating the expression levels of downstream antioxidant genes.

Bacillus Subtilis Delays Neurodegeneration and Behavioral Impairment in the Alzheimer's Disease Model Caenorhabditis Elegans.

Multiple causes, apart from genetic inheritance, predispose to the production and aggregation of amyloid-β (Aβ) peptide and Alzheimer's disease (AD) development in the older population. There is currently no therapy or medicine to prevent or delay AD progression. One novel strategy against AD might involve the use of psychobiotics, probiotic gut bacteria with specific mental health benefits. Here, we report the neuronal and behavioral protective effects of the probiotic bacterium Bacillus subtilis in a Caenorhabditis elegans AD model. Aging and neuronal deterioration constitute important risk factors for AD development, and we showed that B. subtilis significantly delayed both detrimental processes in the wild-type C. elegans strain N2 compared with N2 worms colonized by the non-probiotic Escherichia coli OP50 strain. Importantly, B. subtilis alleviated the AD-related paralysis phenotype of the transgenic C. elegans strains CL2120 and GMC101 that express, in body wall muscle cells, the toxic peptides Aβ3-42 and Aβ1-42, respectively. B. subtilis-colonized CL2355 worms were protected from the behavioral deficits (e.g., poor chemotactic response and decreased body bends) produced by pan-neuronal Aβ1-42 expression. Notably, B. subtilis restored the lifespan level of C. elegans strains that express Aβ to values similar to the life expectancy of the wild-type strain N2 fed on E. coli OP50 cells. The B. subtilis proficiencies in quorum-sensing peptide (i.e., the Competence Sporulation Factor, CSF) synthesis and gut-associated biofilm formation (related to the anti-aging effect of the probiotic) play a crucial role in the anti-AD effects of B. subtilis. These novel results are discussed in the context of how B. subtilis might exert its beneficial effects from the gut to the brain of people with or at risk of developing AD.

Xanthigen® reduces lipid deposition and improves stress resistance in Caenorhabditis elegans

Xanthigen® is a nutraceutical combination of two well-known natural products, brown seaweed extract (rich in fucoxanthin) and pomegranate seed oil (rich in punicic acid), and it has been designed to use in weight management, in conjunction with a calorie restricted diet. In the nematode Caenorhabditis elegans Xanthigen® treatment caused a significant reduction in lipid deposition in wild-type N2 (WT-N2) animals but not in sirt-2.1-deficient strain, which raises the possibility that the prolipolytic or anti-lipogenic effect of Xanthigen® in these animals is mediated through Sirtuin 2.1 activation. This response has been well described for Xanthigen® in cell cultures and other animal models. In addition, Xanthigen® treatment conferred to both strains an increased resistance to thermal and oxidative stress, which opens the possibility that the effects of Xanthigen® are not mediated solely by Sirtuin 2.1 activation. We therefore explored whether Xanthigen® could activate diverse defence mechanisms such as DAF-16 activation, or GST induction in response to xenobiotics, by using the strains TJ356, CL2070 and CL2166, stably expressing Pdaf-16::GFP, Phsp-16.2::GFP and Pgst-4::GFP, respectively. Xanthigen® treatment provoked neither DAF-16 translocation to the nucleus nor increased expression of HSP16.2 and GST4, which opens the possibility that different mechanisms other than DAF-16 and those involved in xenobiotic responses, are activated by Xanthigen® and are capable of conferring to the nematode an increased resistance to thermal or oxidative stress.

N,N’ bis-(2-mercaptoethyl) isophthalamide induces developmental delay in Caenorhabditis elegans by promoting DAF-16 nuclear localization

N,N’ bis-(2-mercaptoethyl) isophthalamide (NBMI) is a lipophilic thiol-containing agent that has high affinity for toxic metal ions, such as Hg2+, Pb2+, and Cd2+. Studies have shown that NBMI is a potent chelator of heavy metals, yet its potential toxicity in animals has yet to be determined. Using the model organism Caenorhabditis elegans (C. elegans), we show no significant change in worms’ death rate or lifespan following NBMI treatment (10−1000 μM). However, NBMI treatment was associated with a significant developmental delay. To determine if the daf-2/age-1/daf-16 pathway is involved in NBMI toxicity, mRNA levels of these genes were assessed in worms treated with NBMI. Here, we found that while NBMI failed to significantly alter the expression of daf-16 or daf-2; age-1 was significantly downregulated by NBMI. Furthermore, NBMI significantly increased DAF-16 nuclear localization. Consistent with a role for this pathway in NBMI toxicity, the developmental arrest by NBMI was more prominent in the DAF-16 transgenic strain than in the wild type N2 strain. Moreover, in the mutant strains harboring null alleles of daf-16, NBMI had no effect on development. In addition, NBMI repressed the expression of detoxifying genes (skn-1, gst-4 and gcs-1). In summary, NBMI has a low developmental toxicity in the C. elegans model, and the nuclear translocation of DAF-16 is involved in the developmental effect of NBMI.