Li-Jung Tai et al. (1999) International C. elegans Meeting "CeHSF and HSB-1: The heat shock response regulators"

Catherine Airey, Sue Fox, Tim Roytman, Sally McFall, Sanjeev Satyal, Richard Morimoto, Li-Jung Tai

[International C. elegans Meeting, 1999]

One of the key components of the stress response is the heat shock transcription factor whose activation leads to synthesis of heat shock proteins. We have analyzed the C. elegans HSF with the help of C. elegans genome project, and found the following features. First, the C. elegans genome appears to encode a single heat shock factor (CeHSF Y53C10A.12), unlike higher metazoans whose genomes encode multiple HSFs. Second, CeHSF encodes a 75 KD protein with a DNA binding domain (residues 87 to 196) and a trimerization domain composed of hydrophobic repeats (HR) A and B (residues 193 to 285). Although the overall identities of CeHSF to S. Cerevisiae (ScHSF) and human HSF (HsHSF) are 17 and 24% respectively, the identities of the DNA binding and trimerization domains to ScHSF and HsHSF are 23 and 40% respectively. Third, sequence analysis of CeHSF showed an apparent lack of HR-C, a regulatory motif of heptad repeats at the C terminus conserved in all known animal HSFs. HR-C has been shown to be important for keeping HSF in a latent monomeric state during unstressed condition. The fact that C. elegans heat shock response has been shown to be inducible, yet CeHSF lacks the HR-C to keep it inactive in normal condition indicates that the mechanism of CeHSF activation may be distinct from other known animal HSFs. Lastly, the EST of C. elegans HSF (EST yk485h11.5) binds specifically to the consensus heat shock element when expressed as a recombinant fusion protein in bacteria. Currently, we are making antibodies and deletion worms to study the expression and function of CeHSF. In the process of dissecting the regulatory mechanism of HSF, a heat shock binding protein (HSB-1) was cloned and shown to negatively regulate the heat shock response in both C. elegans and mammalian cell lines. The HSB-1 deletion worms have a 30% decrease in life span comparing to N2 wildtype worms. The role of HSB-1 in regulating the heat shock response and its interaction with HSF are being examined using overexpression and deletion worms, as well as mammalian tissue culture models. Satyal SH et. al. Genes and Dev. 1998, 12(13):1962-74. Stringham, EG et. al. Molecular Biology of the Cell. 1992, 3:221-233. Wu, C. Annu. Rev. Cell Dev. Biol. 1995, 11: 441-69.

Chen J et al. (2016) Food Funct "Extracts of Tsai Tai (Brassica chinensis): enhanced antioxidant activity and anti-aging effects both in vitro and in Caenorhabditis elegans."

Xiang L, Liu X, Huang Z, Zhou X, Xiang Y, Zhang J, Chen J, Liu Y, He X

[Food Funct, 2016]

Tsai Tai is one of the most widely consumed Brassica vegetables in Asian countries because of its good taste and its nutritional benefits. This study evaluated the antioxidant capacity and possible associated health benefits of 3 Tsai Tai (Brassica chinensis) varieties, namely, Hon Tsai Tai, Pak Choi and Choi Sum. The DPPH radical scavenging ability and reducing power assays were performed to evaluate the in vitro activities of the extracts. Caenorhabditis elegans was used as an in vivo model for evaluation of beneficial health effects, including antioxidant activity and delayed aging. In vitro, the Hon Tsai Tai extract exhibited higher antioxidant activities than Pak Choi and Choi Sum, and the total phenolic contents were significantly correlated with the DPPH and RP values. In vivo, the three assayed Tsai Tai extracts significantly increased resistance against paraquat-induced oxidative stress with an increase in survival rates from 15% to 28% compared with controls. However, only the extract from Hon Tsai Tai significantly prolonged the lifespan of Caenorhabditis elegans, with an 8% increase in the mean lifespan with respect to controls. Further evidence of antioxidant protection was obtained by assessing ROS production via the DCF assay. The analyses of intracellular SOD activity and MDA content confirmed the existence of an antioxidant protective effect. These results suggest that Tsai Tai might serve as a good source of natural antioxidants, and in particular, Hon Tsai Tai could be explored as a potential dietary supplement to retard aging.

Gopal E et al. (2015) J Pharmacol Exp Ther "Species-specific influence of lithium on the activity of SLC13A5 (NaCT): lithium-induced activation is specific for the transporter in primates."

Ganapathy V, Ramachandran S, Gopal E, Babu E, Prasad PD, Bhutia YD

[J Pharmacol Exp Ther, 2015]

NaCT (SLC13A5) is a Na(+)-coupled transporter for Krebs cycle intermediates and is expressed predominantly in the liver. Human NaCT is relatively specific for citrate compared with other Krebs cycle intermediates. The transport activity of human NaCT is stimulated by Li(+), whereas that of rat NaCT is inhibited by Li(+). We studied the influence of Li(+) on NaCTs cloned from eight different species. Li(+) stimulated the activity of only NaCTs from primates (human, chimpanzee, and monkey); by contrast, NaCTs from nonprimate species (mouse, rat, dog, and zebrafish) were inhibited by Li(+). Caenorhabditis elegans NaCT was not affected by Li(+). With human NaCT, the Li(+)-induced increase in transport activity was associated with the conversion of the transporter from a low-affinity/high-capacity type to a high-affinity/low-capacity type. H(+) was able to substitute for Li(+) in eliciting the stimulatory effect. The amino acid Phe500 in human NaCT was critical for Li(+)/H(+)-induced stimulation. Mutation of this amino acid to tryptophan (F500W) markedly increased the basal transport activity of human NaCT in the absence of Li(+), but the ability of Li(+) to stimulate the transporter was almost completely lost with this mutant. Substitution of Phe500 with tryptophan in human NaCT converted the transporter from a low-affinity/high-capacity type to a high-affinity/low-capacity type, an effect similar to that of Li(+) on the wild-type NaCT. These studies show that Li(+)-induced activation of NaCT is specific for the transporter in primates and that the region surrounding Phe500 in primate NaCTs is important for the Li(+) effect.

Tabuse Y (1997) International C. elegans Meeting "LITHIUM CAUSES DEVELOPMENTAL DISORDER IN C. ELEGANS"

Tabuse Y

[International C. elegans Meeting, 1997]

Lithium (Li) has long been known to have teratogenic effects on the development of many organisms, including sea urchin, Xenopus and Dictyostelium. In Li-treated Xenopus embryos, ventral blastmeres are respecified to develop into dorsal structures, leading to dorsalized embryos lacking ventral mesodermal tissues. In Dictyostelium, Li alters the fate of prespore cells to become prestalk cells instead. Besides teratogenic effects, Li is also known to be a most effective treatment of manic-depressive illness.!@Although several models have been proposed to explain Li action, the molecular mechanism has remained unclear. Recently, GSK (glycogen synthase kinase)-3b was proposed to be a target of Li, suggesting that Li affects the wnt signaling pathway. To understand the mechanism of Li action, I first examined the effect of Li on C. elegans embryogenesis. I inoculated N2 animals at the late L4 stage onto NG plates containing 20 mM LiCl, incubated them at 20 oC. The number of eggs produced by treated animals was reduced to about half of the untreated control. Although cell division seemed to proceed, no embryos hatched on Li plates. Treated-embryos developed to produce gut granules, but did not execute normal morphogenesis at later embryonic stages. To identify genes involved in the action of Li, I have begun to screen for Li-resistant mutants that propagated on Li-containing medium. Several mutants were isolated, and one of them was mapped on the left side of unc-42 on chromosome V. On Li plates, the hatching rate of mutant eggs cross-fertilized by wild-type males was essentially the same as that for self-fertilized mutant eggs. On the contrary, no wild-type eggs cross-fertilized by mutant males hatched on Li-containing plates. So, this mutation appeared to be maternal. Further genetic analyses of the mutants and the observation on the cellular phenotype of Li-treated embryos are underway.

Tabuse Y (1996) Worm Breeder's Gazette "Lithium arrests embryonic development of C. elegans"

Tabuse Y

[Worm Breeder's Gazette, 1996]

Lithium (Li) has long been known to have teratogenic effects on the development of many organisms, including sea urchin, Xenopus and Dictyostelium. In Li-treated Xenopus embryos, ventral blastmeres are respecified to develop into dorsal structures, leading to dorsalized embryos lacking ventral mesodermal tissues. In Dictyostelium, Li alters the fate of prespore cells to become prestalk cells instead. Besides teratogenic effects, Li is also known to bea most effective treatment of manic-depressive illness. Although several models have been proposed to explain Li action, the molecular mechansm remains unclear. The most widely accepted model is the inositol depletion hypothesis, in which Li is thought to affect inositol phosphate turnover by inhibiting inositol monophosphatase, thus resulting in the depletion of endogenous inositol. To understand the mechanism of Li action, I first examined the effect of Li on C. elegans embryogenesis. I inoculated N2 animals at the late L4 stage onto NG plates containing 20 mM LiCl, incubated them at 20 C and observed the laid embryos. The number of eggs produced by treated animals was reduced to about half of the untreated control. Although cell division seemed to proceed, no embryos hatched on Li plates. Treated-embryos developed to produce gut granules, but did not execute normal morphogenesis at later embryonic stages. To identify genes involved in the action of Li, I have begun to screen for Li-resistant mutants, which propagated on Li-containing medium. So far, I obtained one mutant. The mutation was tentatively assigned to chromosome V. Preliminary genetic analysis showed that the mutation showed maternal effect. On Li plates, the hatching rate of mutant eggs cross-fertilized by wild-type males was essentially the same as that for self-fertilized mutant eggs. On the contrary, no wild-type eggs cross-fertilized by mutant males hatched on Li-containing plates. I am now trying to isolate other mutants and also to identify early defects of embryogenesis caused by lithium treatment. I would like to thank J. Miwa for encouragement and discussions.

Inokuchi A et al. (2015) J Appl Toxicol "Effects of lithium on growth, maturation, reproduction and gene expression in the nematode Caenorhabditis elegans."

Matsusaki H, Yamamoto R, Takumi S, Morita F, Inokuchi A, Arizono K, Tominaga N, Ishibashi H

[J Appl Toxicol, 2015]

Lithium (Li) has been widely used to treat bipolar disorder, and industrial use of Li has been increasing; thus, environmental pollution and ecological impacts of Li have become a concern. This study was conducted to clarify the potential biological effects of LiCl and Li(2)CO(3) on a nematode, Caenorhabditis elegans as a model system for evaluating soil contaminated with Li. Exposure of C. elegans to LiCl and Li(2)CO(3) decreased growth/maturation and reproduction. The lowest observed effect concentrations for growth, maturation and reproduction were 1250, 313 and 10 000m, respectively, for LiCl and 750, 750 and 3000m, respectively, for Li(2)CO(3). We also investigated the physiological function of LiCl and LiCO(3) in C. elegans using DNA microarray analysis as an eco-toxicogenomic approach. Among approximately 300 unique genes, including metabolic genes, the exposure to 78m LiCl downregulated the expression of 36 cytochrome P450, 16 ABC transporter, 10 glutathione S-transferase, 16 lipid metabolism and two vitellogenin genes. On the other hand, exposure to 375m Li(2)CO(3) downregulated the expression of 11 cytochrome P450, 13 ABC transporter, 13 lipid metabolism and one vitellogenin genes. No gene was upregulated by LiCl or Li(2)CO(3). These results suggest that LiCl and Li(2)CO(3) potentially affect the biological and physiological function in C. elegans associated with alteration of the gene expression such as metabolic genes. Our data also provide experimental support for the utility of toxicogenomics by integrating gene expression profiling into a toxicological study of an environmentally important organism such as C. elegans.

McColl G et al. (2008) J Biol Chem "Pharmacogenetic analysis of lithium-induced delayed aging in Caenorhabditis elegans."

Melov S, McColl G, Lithgow GJ, Vantipalli MC, Killilea DW, Hubbard AE

[J Biol Chem, 2008]

Lithium (Li+) has been used to treat mood affect disorders, including bipolar, for decades (1;2). This drug is neuroprotective and has several identified molecular targets. However, it has a narrow therapeutic range and the underlying mechanism(s) of its therapeutic action is not understood. Here we describe a pharmacogenetic study of Li+ in the nematode Caenorhabditis elegans. Exposure to Li+ at clinically relevant concentrations throughout adulthood increases survival during normal aging (up to 46% median increase). Longevity is extended via a novel mechanism with altered expression of genes encoding nucleosome-associated functions. Li+ treatment results in reduced expression of the worm ortholog of LSD-1 (T08D10.2), a histone demethylase; knockdown by RNA interference (RNAi) of T08D10.2 is sufficient to extend longevity (~25% median increase), suggesting Li+ regulates survival by modulating histone methylation and chromatin structure.

D Royal et al. (2001) International C. elegans Meeting "Towards the Cloning of Gene That Can Mutate to Confer Lithium Resistance In C. elegans"

J White, M Driscoll, E Hsieh, N Sirotin, M A Royal, D Royal, S Kwok, K O'Connell, B Bowerman

[International C. elegans Meeting, 2001]

Little is known about how ions permeate the C. elegans gut and cuticle. How various ions influence development and behavior is also not understood. One ion with considerable impact in human biology is Li + , which modulates bipolar disorder by an unknown mechanism. In particular, the targets of lithium that cause side effects remain to be identified. We are using C. elegans to genetically identify targets of lithium and to elaborate on mechanisms of transport, which may have an impact on human health. Li + has dosage-dependent effects on C. elegans embryos. When L4 animals mature on NGM plates with Li + added to the final concentrations of 10mM-20mM, they produce embryos that are unable to hatch. We demonstrated that this failure to hatch is due to defects in cytokinesis that result in multi-nucleated embryos and symmetrically partitioned cells. Li + also has a dosage-dependent effect on larval development. When adult hermaphrodites are permitted to lay embryos on 10mM-20mM Li + NGM plates, the resulting offspring experience a developmental delay proportional to the concentration of Li + . The offspring become progressively paralyzed as they reach adulthood. We demonstrated earlier that there is a delay in the entry into the S phase of the cell cycle larval stages. To learn more about the biology of Li + sensitivity, we screened for Li + resistant mutants. We developed three screens that took advantage of the embryonic arrest and the larval delay caused by Li + . We isolated one mutant bz71 , in a screen of 44,000 haploid genomes, that is resistant to both the larval and embryonic blocks of 16mM Li + . bz71 is dominant. Using classical mapping techniques, we positioned it on LGIII between unc-32 and dpy-18 . We are currently using SNP strategy to obtain a higher resolution map and we hope to report on the identification and cloning of this locus.

James F Morley et al. (2001) International C. elegans Meeting "HEAT SHOCK TRANSCRIPTION FACTOR (HSF) ACTIVITY REGULATES THE APPEARANCE OF POLYGLUTAMINE PROTEIN AGGREGATES"

Li-Jung Tai, Richard I Morimoto, James F Morley

[International C. elegans Meeting, 2001]

Disruption of protein folding homeostasis resulting in protein misfolding and aggregation is associated with many human diseases including Huntington's disease where expansion of a polyglutamine (polyQ) tract to greater than 37-40 residues leads to the formation of protein aggregates, neuronal dysfunction and premature cell death. The expression of expanded chimeric polyglutamine (polyQ) -YFP proteins as extrachromosomal arrays in body wall muscle cells results in the formation of visible protein aggregates associated with a10-fold reduction in motility. Immunohistochemical studies of Q82-expressing animals reveal displacement of actin and myosin filaments. In contrast, neither visible aggregates nor altered motility were detected in animals expressing Q19 or Q29-YFP. Animals expressing an intermediate polyQ tract (Q40) exhibit striking polymorphism, as different muscle cells in the same animal contain soluble or aggregated polyQ. The severity of motility defect in Q40 animals corresponds to the proportion of cells with aggregates. Our laboratory and others have previously identified individual molecular chaperones as modulators of the aggregation phenotype. As chaperones are co-regulated during the heat shock response by HSF, we have examined whether altering the levels of HSF by RNAi and tissue-specific overexpression would impact the biochemical events that influence protein aggregation. Expression of a full length Ce-hsf cDNA from the unc-54 promoter results in more rapid activation of an hsp-16::lacZ reporter in body wall muscle cells, confirming overexpression of functional HSF. In unc-54::Q82-yfp; myo-2::Q82-gfp; unc-54::hsf animals, we observe the appearance of soluble patches of fluorescence in body wall muscle, but not in the pharynx. However, expression of a C-terminal HSF fragment that lacks DNA binding and trimerization domains has no effect on activation of the heat shock response or distribution of Q82-YFP. Injection of dsRNA transcribed from a full-length hsf cDNA inhibits activation of the heat shock response in heat shocked hsp-16::lacZ animals, confirming inactivation of HSF by RNAi. Inhibition of HSF in Q19 or Q29 animals leads to sporadic formation of aggregates similar to those observed in Q82 strains. Consistent with the transient nature of RNAi phenotypes, F1 progeny of injected animals were affected, but F2 animals exhibit normal activation of the heat shock response and distribution of Q19. Normal activation of the heat shock response and distribution of Q19 in animals injected with unc-22 dsRNA demonstrated the specificity of hsf RNAi effects. These data suggest that Q19 and Q29 solubility is maintained by an active process that requires HSF activity. The heat shock response is considered an acute response to changes in environmental conditions, and the requirement for HSF activity to maintain Q19 and Q29 solubility in the absence of stress reveals a novel role for HSF. We are currently testing whether manipulations of HSF levels that influence aggregate formation are associated with altered motility.

Li WQ et al. (1992) Worm Breeder's Gazette "Characterization of the Axonal Guidance and Outgrowth gene Unc-33"

Shaw JE, Li WQ, Herman RK

[Worm Breeder's Gazette, 1992]

Characterization of the axonal guidance and outgrowth gene unc-33 W. Li, R. K. Herman and J. E. Shaw Department of Genetics and Cell biology, University of Minnesota, St Paul, MN 55108