Butcher RA (2017) Nat Prod Rep "Decoding chemical communication in nematodes."

Butcher RA

[Nat Prod Rep, 2017]

The nematode Caenorhabditis elegans produces tens, if not hundreds, of different ascarosides as pheromones to communicate with other members of its species. Overlapping mixtures of these pheromones affect the development of the worm and a variety of different behaviors. The ascarosides represent a unique tool for dissecting the neural circuitry that controls behavior and that connects to important signaling pathways, such as the insulin and TGF pathways, that lie at the nexus of development, metabolism, and lifespan in C. elegans. However, the exact physiological roles of many of the ascarosides are unclear, especially since many of these pheromones likely have multiple functions depending on their concentrations, the presence of other pheromones, and a variety of other factors. Determining these physiological roles will be facilitated by top-down approaches to characterize the pheromone receptors and their function, as well as bottom-up approaches to characterize the pheromone biosynthetic enzymes and their regulation.

Butcher RA (2017) Nat Chem Biol "Small-molecule pheromones and hormones controlling nematode development."

Butcher RA

[Nat Chem Biol, 2017]

The existence of small-molecule signals that influence development in Caenorhabditis elegans has been known for several decades, but only in recent years have the chemical structures of several of these signals been established. The identification of these signals has enabled connections to be made between these small molecules and fundamental signaling pathways in C. elegans that influence not only development but also metabolism, fertility, and lifespan. Spurred by these important discoveries and aided by recent advances in comparative metabolomics and NMR spectroscopy, the field of nematode chemistry has the potential to expand dramatically in the coming years. This Perspective will focus on small-molecule pheromones and hormones that influence developmental events in the nematode life cycle (ascarosides, dafachronic acids, and nemamides), will cover more recent work regarding the biosynthesis of these signals, and will explore how the discovery of these signals is transforming our understanding of nematode development and physiology.

Butcher RA et al. (2009) Proc Natl Acad Sci U S A "Biosynthesis of the Caenorhabditis elegans dauer pheromone."

Mak HY, Butcher RA, Clardy J, Li W, Ragains JR, Ruvkun G

[Proc Natl Acad Sci U S A, 2009]

To sense its population density and to trigger entry into the stress-resistant dauer larval stage, Caenorhabditis elegans uses the dauer pheromone, which consists of ascaroside derivatives with short, fatty acid-like side chains. Although the dauer pheromone has been studied for 25 years, its biosynthesis is completely uncharacterized. The daf-22 mutant is the only known mutant defective in dauer pheromone production. Here, we show that daf-22 encodes a homolog of human sterol carrier protein SCPx, which catalyzes the final step in peroxisomal fatty acid beta-oxidation. We also show that dhs-28, which encodes a homolog of the human d-bifunctional protein that acts just upstream of SCPx, is also required for pheromone production. Long-term daf-22 and dhs-28 cultures develop dauer-inducing activity by accumulating less active, long-chain fatty acid ascaroside derivatives. Thus, daf-22 and dhs-28 are required for the biosynthesis of the short-chain fatty acid-derived side chains of the dauer pheromone and link dauer pheromone production to metabolic state.

Butcher RA et al. (2007) Nat Chem Biol "Small-molecule pheromones that control dauer development in Caenorhabditis elegans."

Schroeder FC, Fujita M, Butcher RA, Clardy J

[Nat Chem Biol, 2007]

In response to high population density or low food supply, the nematode Caenorhabditis elegans enters an alternative larval stage, known as the dauer, that can withstand adverse conditions for prolonged periods. C. elegans senses its population density through a small-molecule signal, traditionally called the dauer pheromone, that it secretes into its surroundings. Here we show that the dauer pheromone consists of several structurally related ascarosides-derivatives of the dideoxysugar ascarylose-and that two of these ascarosides (1 and 2) are roughly two orders of magnitude more potent at inducing dauer formation than a previously reported dauer pheromone component (3) and constitute a physiologically relevant signal. The identification of dauer pheromone components 1 and 2 will facilitate the identification of target receptors and downstream signaling proteins.

Butcher RA et al. (2009) Org Lett "An indole-containing dauer pheromone component with unusual dauer inhibitory activity at higher concentrations."

Ragains JR, Clardy J, Butcher RA

[Org Lett, 2009]

In Caenorhabditis elegans, the dauer pheromone, which consists of a number of derivatives of the 3,6-dideoxysugar ascarylose, is the primary cue for entry into the stress-resistant, "nonaging" dauer larval stage. Here, using activity-guided fractionation and NMR-based structure elucidation, a structurally novel, indole-3-carboxyl-modified ascaroside is identified that promotes dauer formation at low nanomolar concentrations but inhibits dauer formation at higher concentrations.

Butcher RA et al. (2008) Proc Natl Acad Sci U S A "A potent dauer pheromone component in Caenorhabditis elegans that acts synergistically with other components."

Ragains JR, Kim E, Butcher RA, Clardy J

[Proc Natl Acad Sci U S A, 2008]

In the model organism Caenorhabditis elegans, the dauer pheromone is the primary cue for entry into the developmentally arrested, dauer larval stage. The dauer is specialized for survival under harsh environmental conditions and is considered "nonaging" because larvae that exit dauer have a normal life span. C. elegans constitutively secretes the dauer pheromone into its environment, enabling it to sense its population density. Several components of the dauer pheromone have been identified as derivatives of the dideoxy sugar ascarylose, but additional unidentified components of the dauer pheromone contribute to its activity. Here, we show that an ascaroside with a 3-hydroxypropionate side chain is a highly potent component of the dauer pheromone that acts synergistically with previously identified components. Furthermore, we show that the active dauer pheromone components that are produced by C. elegans vary depending on cultivation conditions. Identifying the active components of the dauer pheromone, the conditions under which they are produced, and their mechanisms of action will greatly extend our understanding of how chemosensory cues from the environment can influence such fundamental processes as development, metabolism, and aging in nematodes and in higher organisms.

Harris TW et al. (2000) J Cell Biol "Mutations in synaptojanin disrupt synaptic vesicle recycling."

Harris TW, Hartweig EA, Jorgensen EM, Horvitz HR

[J Cell Biol, 2000]

Synaptojanin is a polyphosphoinositide phosphatase that is found at synapses and binds to proteins implicated in endocytosis. For these reasons, it has been proposed that synaptojanin is involved in the recycling of synaptic vesicles. Here, we demonstrate that the unc-26 gene encodes the Caenorhabditis elegans ortholog of synaptojanin. unc-26 mutants exhibit defects in vesicle trafficking in several tissues, but most defects are found at synaptic termini. Specifically, we observed defects in the budding of synaptic vesicles from the plasma membrane, in the uncoating of vesicles after fission, in the recovery of vesicles from endosomes, and in the tethering of vesicles to the cytoskeleton. Thus, these results confirm studies of the mouse synaptojanin 1 mutants, which exhibit defects in the uncoating of synaptic vesicles (Cremona, O., G. Di Paolo, M.R. Wenk, A. Luthi, W.T. Kim, K. Takei, L. Daniell, Y. Nemoto, S.B. Shears, R.A. Flavell, D.A. McCormick, and P. De Camilli. 1999. Cell. 99:179-188), and further demonstrate that synaptojanin facilitates multiple steps of synaptic vesicle recycling.

Benedetti MG et al. (2008) Exp Gerontol "Compounds that confer thermal stress resistance and extended lifespan."

Benedetti MG, Lithgow GJ, White MP, Sampayo JN, Foster AL, Vantipalli MC, Gill MS, Olsen A

[Exp Gerontol, 2008]

The observation that long-lived and relatively healthy animals can be obtained by simple genetic manipulation prompts the search for chemical compounds that have similar effects. Since aging is the most important risk factor for many socially and economically important diseases, the discovery of a wide range of chemical modulators of aging in model organisms could prompt new strategies for attacking age-related disease such as diabetes, cancer and neurodegenerative disorders [Collins, J.J., Evason, K., Kornfeld, K., 2006. Pharmacology of delayed aging and extended lifespan of Caenorhabditis elegans. Exp. Gerontol.; Floyd, R.A., 2006. Nitrones as therapeutics in age-related diseases. Aging Cell 5, 51-57; Gill, M.S., 2006. Endocrine targets for pharmacological intervention in aging in Caenorhabditis elegans. Aging Cell 5, 23-30; Hefti, F.F., Bales, R., 2006. Regulatory issues in aging pharmacology. Aging Cell 5, 3-8]. Resistance to multiple types of stress is a common trait in long-lived genetic variants of a number of species; therefore, we have tested compounds that act as stress response mimetics. We have focused on compounds with antioxidant properties and identified those that confer thermal stress resistance in the nematode Caenorhabditis elegans. Some of these compounds (lipoic acid, propyl gallate, trolox and taxifolin) also extend the normal lifespan of this simple invertebrate, consistent with the general model that enhanced stress resistance slows aging.

Reilly DK et al. (2019) MicroPubl Biol "Evolution of hermaphroditism decreases efficacy of Ascaroside#8-mediated mate attraction in Caenorhabditis nematodes"

Srinivasan J, Randle LJ, Reilly DK

[MicroPubl Biol, 2019]

Nematodes, such as the model organism Caenorhabditis elegans, communicate environmental and developmental information with conspecifics through a class of small-molecule pheromones termed ascarosides (Butcher, 2017; Chute and Srinivasan, 2014; Ludewig and Schroeder, 2013). Nematodes share ascaroside signaling pathways (Choe et al., 2012), but are also capable of eavesdropping on chemical signals of predatory species (Liu et al., 2018). Ascarosides signal vast arrays of information, either individually or as blends, based on concentration, sex, physiological state, and other ascarosides sensed (McGrath and Ruvinsky, 2019; Pungaliya et al., 2009; Srinivasan et al., 2008; Srinivasan et al., 2012). For instance, octopamine-succinylated ascaroside #9 (osas#9) is able to signal starvation conditions in the absence of other ascarosides (Artyukhin et al., 2013).C. elegans (Cel) is an androdioecious species, with the majority of the natural population comprised of self-fertilizing hermaphrodites, and a small proportion (<0.2%) being male (Hodgkin et al., 1979). There are two other similarly androdioecious species in the genus, C. briggsae (Cbr) and C. tropicalis (Ctr). All three species evolved their hermaphroditism separately and uniquely (Ellis and Lin, 2014). Of the male-attracting ascarosides secreted by C. elegans (ascr#2, ascr#3, ascr#4, and ascr#8), ascr#8 is the most potent (Pungaliya et al., 2009). Since ascr#8 is a male attractant in this hermaphroditic species, we asked if other hermaphroditic species retained the ability to attract males using this cue. Males from the gonochoristic (male-female) sister species to C. briggsae and C. tropicalis C. nigoni (Cni) and C. wallacei (Cwa), respectively were also assayed for their ability to respond to ascr#8. The closest relative of C. elegans, the gonochoristic C. inopinata (Cin, formerly C. sp. 34), which has been recently characterized (Kanzaki et al., 2018), was also tested, along with the JaponicaGroup gonochoristic species C. japonica(Cja) and C. afra(Caf).Dwell times were analyzed as previously described using a Spot Retention Assay (Narayan et al., 2016). Dwell times were transformed using a Base 2 Exponentiation (2n, wherein n is equal to the raw dwell time value) to generate only non-zero data in order to calculate fold-changes. The Logbase2 of the fold-changes was then calculated to normalize the data. All data sets were first checked for normality using a DAgostino