Miller D L et al. (2010) Aging, Metabolism, Stress, Pathogenesis, and Small RNAs, Madison, WI "Hydrogen Sulfide Protects Against Hypoxia Induced Protein Aggregation"

Miller D L, Roth M B

[Aging, Metabolism, Stress, Pathogenesis, and Small RNAs, Madison, WI, 2010]

With age, the ability to maintain protein homeostasis declines. We have shown previously that low levels of hydrogen sulfide (H2S) increases lifespan and thermotolerance in the nematode C. elegans. Here we show that H2S also improves the ability to maintain protein homeostasis, as judged by the aggregation of polyglutamine (polyQ) containing proteins. Our data suggest that adaptation to H2S impinges on protein homeostasis through a mechanism that is distinct from the effects of H2S on lifespan. Remarkably, we have found that H2S also improves the ability to maintain protein homeostasis in stressful environmental conditions. We demonstrate that polyQ proteins aggregate in vivo when animals are exposed to specific hypoxic environments. The range of O2 concentrations that induce protein aggregation is modulated by genetic factors, including hif-1. Adaptation to H2S protects against hypoxia-induced protein aggregation, as animals pre-treated with H2S develop fewer protein aggregates when exposed to hypoxia. We have also observed that treatment with H2S after the hypoxic insult retards subsequent polyQ protein aggregation. These experiments show that polyQ-protein aggregation is induced both during and after exposure to hypoxia, which may have important clinical implications. Recently, it has been demonstrated that HIF-1 protein is stabilized and activated by H2S (Budde and Roth, 2010), suggesting the possibility that H2S acts through HIF-1 to modulate protein quality control mechanisms. Together, our studies show that H2S can have remarkably long-lasting physiological effects that can improve homeostatic mechanisms required to appropriately respond to subsequence environmental perturbations.

D Miller et al. (1999) International C. elegans Meeting "Two-color GFP expression in C. elegans"

D Piston, D Hardin, A Fire, J Fleenor, G Patterson, D Miller, N Desai

[International C. elegans Meeting, 1999]

The Green Fluorescent Protein (GFP) is now widely used for imaging cells and organelles in living animals. GFP variants with distinct excitation and emission spectra have been developed and should prove highly useful for simultaneous imaging of separate proteins labeled with different colored GFPs. Here we describe expression vectors and fluorescence filter sets that can be used to obtain bright, distinct images of "Cyan" GFP (CFP) and "Yellow" GFP (YFP) in C. elegans (1). GFP vectors specifically designed for expression in C. elegans (2) were modified to include amino acid substitutions appropriate for CFP (Y66W, N146I, M153T, V163A) or YFP (S65G, V68A, S72A, T203Y) (see www.ciwemb.edu ). Previously described promoter regions and localization signals were employed to create trangenic animals expressing CFP and YFP either in different cells or in separate intracellular compartments. In these experiments, CFP appears "Blue" and the YFP signal is "Green." For example, an unc-4 promoter element drives expression in VA motor neurons whereas a del-1 upstream region is specific for the adjacent VB motor neurons. Transgenic animals expressing both unc-4::CFP and del-1::YFP display side-by-side VA (Blue) and VB (Green) motor neurons in the ventral nerve cord. In another experiment, cytoplasmically localized YFP and nuclear-localized CFP were expressed under the control of the muscle-specific unc-54 promoter to produce Green bodywall muscle cells with Blue nuclei. It should now be possible to utilize these vectors for a wide variety of two-color labeling experiments. The fluorescence filter sets used in the work were developed in collaboration with Chroma Technology Corp. For CFP: excitation = 436/10 nm; dichroic = 450 nm; emission = 485/50 nm For YFP: excitation = 500/20 nm; dichroic = 515 nm; emission = 520 nm long pass. (1) Miller, et al. (1999) BioTechniques, in press. (2) Fire, et al. (1998). p. 153-168. In M. Chalfie and S. Kain (Eds.), GFP Strategies and Applications. John Wiley and Sons, NY

Johnson, Christina K. et al. (2021) MicroPubl Biol

Miller III, David M., Johnson, Christina K., Bianchi, Laura

[MicroPubl Biol, 2021]

For Johnson, CK; Miller, DD; Bianchi, L (2021). Effect of the protease plasmin on C. elegans hyperactive DEG/ENaC channels MEC-4(d) and UNC-8(d). microPublication Biology. 10.17912/micropub.biology.000412.

Miller DM et al. (1992) Worm Breeder's Gazette "unc-4 LacZ Expression in A-Type Motor Neurons"

Miller DM, Niemeyer CJ

[Worm Breeder's Gazette, 1992]

unc-4 LacZ expression in A-type motor neurons David M. Miller and Charles J. Niemeyer, Dept. of Cell Biology, Duke Univ. Medical Ctr, Durham, NC 27710

Sakoguchi H et al. (2016) Biosci Biotechnol Biochem "Screening of biologically active monosaccharides: growth inhibitory effects of d-allose, d-talose, and l-idose against the nematode Caenorhabditis elegans."

Izumori K, Yoshihara A, Sakoguchi H, Sato M

[Biosci Biotechnol Biochem, 2016]

We compared the growth inhibitory effects of all aldohexose stereoisomers against the model animal Caenorhabditis elegans. Among the tested compounds, the rare sugars d-allose (d-All), d-talose (d-Tal), and l-idose (l-Ido) showed considerable growth inhibition under both monoxenic and axenic culture conditions. 6-Deoxy-d-All had no effect on growth, which suggests that C6-phosphorylation by hexokinase is essential for inhibition by d-All.

Sakoguchi H et al. (2016) Bioorg Med Chem Lett "Growth inhibitory effect of d-arabinose against the nematode Caenorhabditis elegans: Discovery of a novel bioactive monosaccharide."

Shintani T, Izumori K, Sato M, Sakoguchi H, Okuma K, Yoshihara A

[Bioorg Med Chem Lett, 2016]

Biological activities of unusual monosaccharides (rare sugars) have largely remained unstudied until recently. We compared the growth inhibitory effects of aldohexose stereoisomers against the animal model Caenorhabditis elegans cultured in monoxenic conditions with Escherichia coli as food. Among these stereoisomers, the rare sugar d-arabinose (d-Ara) showed particularly strong growth inhibition. The IC50 value for d-Ara was estimated to be 7.5mM, which surpassed that of the potent glycolytic inhibitor 2-deoxy-d-glucose (19.5mM) used as a positive control. The inhibitory effect of d-Ara was also observed in animals cultured in axenic conditions using a chemically defined medium; this excluded the possible influence of E. coli. To our knowledge, this is the first report of biological activity of d-Ara. The d-Ara-induced inhibition was recovered by adding either d-ribose or d-fructose, but not d-glucose. These findings suggest that the inhibition could be induced by multiple mechanisms, for example, disturbance of d-ribose and d-fructose metabolism.

Yoshihara A et al. (2019) Bioorg Med Chem Lett "Growth inhibition by 1-deoxy-d-allulose, a novel bioactive deoxy sugar, screened using Caenorhabditis elegans assay."

Sakoguchi H, Fleet GWJ, Izumori K, Shintani T, Sato M, Yoshihara A

[Bioorg Med Chem Lett, 2019]

The biological activities of deoxy sugars (deoxy monosaccharides) have remained largely unstudied until recently. We compared the growth inhibition by all 1-deoxyketohexoses using the animal model Caenorhabditis elegans. Among the eight stereoisomers, 1-deoxy-d-allulose (1d-d-Alu) showed particularly strong growth inhibition. The 50% inhibition of growth (GI₅₀) concentration by 1d-d-Alu was estimated to be 5.4mM, which is approximately 10 times lower than that of d-allulose (52.7mM), and even lower than that of the potent glycolytic inhibitor, 2-deoxy-d-glucose (19.5mM), implying that 1d-d-Alu has a strong growth inhibition. In contrast, 5-deoxy- and 6-deoxy-d-allulose showed no growth inhibition of C. elegans. The inhibition by 1d-d-Alu was alleviated by the addition of d-ribose or d-fructose. Our findings suggest that 1d-d-Alu-mediated growth inhibition could be induced by the imbalance in d-ribose metabolism. To our knowledge, this is the first report of biological activity of 1d-d-Alu which may be considered as an antimetabolite drug candidate.

Katane M et al. (2020) Biochim Biophys Acta Proteins Proteom "Biochemical characterization of d-aspartate oxidase from Caenorhabditis elegans: Its potential use in the determination of free D-glutamate in biological samples."

Katane M, Sekine M, Nakayama K, Homma H, Kuwabara H, Saitoh Y, Miyamoto T

[Biochim Biophys Acta Proteins Proteom, 2020]

d-Aspartate oxidase (DDO) is a flavin adenine dinucleotide (FAD)-containing flavoprotein that stereospecifically acts on acidic D-amino acids (i.e., free d-aspartate and D-glutamate). Mammalian DDO, which exhibits higher activity toward d-aspartate than D-glutamate, is presumed to regulate levels of d-aspartate in the body and is not thought to degrade D-glutamate in vivo. By contrast, three DDO isoforms are present in the nematode Caenorhabditis elegans, DDO-1, DDO-2, and DDO-3, all of which exhibit substantial activity toward D-glutamate as well as d-aspartate. In this study, we optimized the Escherichia coli culture conditions for production of recombinant C. elegans DDO-1, purified the protein, and showed that it is a flavoprotein with a noncovalently but tightly attached FAD. Furthermore, C. elegans DDO-1, but not mammalian (rat) DDO, efficiently and selectively degraded D-glutamate in addition to d-aspartate, even in the presence of various other amino acids. Thus, C. elegans DDO-1 might be a useful tool for determining these acidic D-amino acids in biological samples.

Shintani T et al. (2019) J Appl Glycosci (1999) "D-Allose, a Stereoisomer of D-Glucose, Extends the Lifespan of Caenorhabditis elegans via Sirtuin and Insulin Signaling."

Izumori K, Sakoguchi H, Yoshihara A, Shintani T, Sato M

[J Appl Glycosci (1999), 2019]

D-Allose (D-All), C-3 epimer of D-glucose, is a rare sugar known to suppress reactive oxygen species generation and prevent hypertension. We previously reported that D-allulose, a structural isomer of D-All, prolongs the lifespan of the nematode Caenorhabditis elegans. Thus, D-All was predicted to affect longevity. In this study, we provide the first empirical evidence that D-All extends the lifespan of C. elegans. Lifespan assays revealed that a lifespan extension was induced by 28 mM D-All. In particular, a lifespan extension of 23.8 % was achieved (p< 0.0001). We further revealed that the effects of D-All on lifespan were dependent on the insulin gene daf-16 and the longevity gene sir-2.1, indicating a distinct mechanism from those of other hexoses, such as D-allulose, with previously reported antiaging effects.

Sato M et al. (2008) J Nat Med "Potential anthelmintic: D-psicose inhibits motility, growth and reproductive maturity of L1 larvae of Caenorhabditis elegans."

Izumori K, Yamasaki T, Kurose H, Sato M

[J Nat Med, 2008]

No anthelmintic sugars have yet been identified. Eight ketohexose stereoisomers (D- and L-forms of psicose, fructose, tagatose and sorbose), along with D-galactose and D-glucose, were examined for potency against L1 stage Caenorhabditis elegans fed Escherichia coli. Of the sugars, D-psicose specifically inhibited the motility, growth and reproductive maturity of the L1 stage. D-Psicose probably interferes with the nematode nutrition. The present results suggest that D-psicose, one of the rare sugars, is a potential anthelmintic.