Ward CJ et al. (2015) Curr Biol "Polycystic Kidney Disease: Lessons Learned from Caenorhabditis elegans Mating Behavior."

Ward CJ, Sharma M

[Curr Biol, 2015]

Male worm mating requires lov-1 and pkd-2 (homologs of the human polycystic kidney disease genes, PKD1and PKD2), which are expressed in male-specific neurons. Transcriptomic analysis of these neuronsnow catalogs molecules involved in signaling and ectosome biogenesis, with implications for human PKD.

Jang M et al. (2020) Antioxidants (Basel) "Antioxidant Capacity of Thistle (Cirsium japonicum) in Various Drying Methods and their Protection Effect on Neuronal PC12 Cells and Caenorhabditis elegans."

Kim GH, Jang M, Kim KH

[Antioxidants (Basel), 2020]

The aim of this study was, firstly, to evaluate the phenol profile of thistle (<i>Cirsium japonicum</i>, CJ) by High performance liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-MS), dried by different methods (90 C hot-air, 70 C hot-air, shade-, and freeze-drying). Secondly, we aimed to evaluate the relationship between phenolic compounds content and antioxidant properties. CJ contained chlorogenic acid, linarin, and pectolinarin. Total phenolic contents of CJ significantly decreased under hot-air-drying condition, especially chlorogenic acid contents in CJ have been reduced by 85% and 60% for 90 C and 70 C hot-air-drying, respectively. We evaluated the protective effect on adrenal pheochromocytoma (PC12) cells and <i>Caenorhabditis elegans</i> using shade-dried CJ, which has the largest phenolic contents and the strongest antioxidant property. CJ-treated PC 12 cells dose-dependently exhibited the protective effects against reactive oxygen species (ROS), while cell viability increases, lactate dehydrogenase release decreases, and ROS formation decreases. Furthermore, CJ has also shown protection against ROS in <i>C. elegans.</i> Consequently, CJ contributed to lifespan extension under ROS stress without influencing the physiological growth.

Williams AB et al. (2010) Mol Cell "Eviction notice: new insights into Rad51 removal from DNA during homologous recombination."

Williams AB, Michael WM

[Mol Cell, 2010]

In this issue of Molecular Cell, Ward et al. (2010) identify two genes whose products act redundantly to clear Rad51 from DNA after successful strand invasion, thereby enabling the downstream events of homologous recombination to go smoothly.

Politi, Kristin A. et al. (2009) International Worm Meeting "Three dimensional structure of the Ward body."

Derr, KD, Rice, William J., Politi, Kristin A., Fisher, Kevin, Crocker, Chris, Hall, David H.

[International Worm Meeting, 2009]

The plasma membrane of the nematode hypodermis features a unique set of stacked membrane invaginations that we presume to be associated with cuticle deposition, although there is still no experimental evidence to prove it. These structures were first noted by TEM in C. elegans by Sam Ward in the early 1970s, and briefly mentioned in WormBook I by John White (1). Here we present electron microscopic evidence for their organization in the wild type adult, combining thin sections, freeze fracture and electron tomography to view their organization in groupings along the body wall. The so-called "Ward body" resembles a Golgi body that has been rotated on end so that each succeeding membrane leaflet can fuse directly to the plasma membrane. Ward bodies are often clustered nearby to one another, but the orientation of each Ward body seems to be independent of its neighbors, and seemingly has no fixed angle with respect to the body axis. In some cases their leaflets lie virtually parallel to the plasma membrane. Similar membrane infoldings have also been seen in the excretory duct''s luminal membrane, but are not known to occur in seam cells or other epithelia in the nematode. We hypothesize that large extended proteins, such as collagens, may co-assemble within the protected environment of a Ward body leaflet prior to deposition in the cuticle. Supported by NIH RR 12596 to DHH. 1. W. Wood (Ed), The nematode C. elegans. Chapter 4, "The Anatomy", Fig 2a, 1988.

Kura T (1999) Trends in Ecology & Evolution "Caenorhabditis and the cost of sex."

Kura T

[Trends in Ecology & Evolution, 1999]

In a recent TREE news & comment, Gadagkar made some useful comments on LaMunyon and Ward's interesting study on sexual reproduction in nematodes. I think, however, that he - and LaMunyon and Ward - have confused the benefits of sex for species or demes with those for individuals or genes.

Kazatskaya, Anna et al. (2020) MicroPubl Biol

de Bono, Mario, Amin-Wetzel, Niko, Sengupta, Piali, Philbrook, Alison, Kazatskaya, Anna, Yuan, Lisa

[MicroPubl Biol, 2020]

A subset of sensory neurons in C. elegans contains compartmentalized sensory structures termed cilia at their distal dendritic ends (Ward et al. 1975; Perkins et al. 1986; Doroquez et al. 2014). Cilia present on different sensory neuron types are specialized both in morphology and function, and are generated and maintained via shared and cell-specific molecules and mechanisms (Perkins et al. 1986; Evans et al. 2006; Mukhopadhyay et al. 2007; Mukhopadhyay et al. 2008; Morsci and Barr 2011; Doroquez et al. 2014; Silva et al. 2017). The bilaterally symmetric pair of URX oxygen-sensing neurons in the C. elegans head (Figure 1A) is thought to be non-ciliated (Ward et al. 1975; Doroquez et al. 2014) but nevertheless exhibits intriguing morphological similarities with ciliated sensory neurons. URX dendrites extend to the nose where they terminate in large bulb-like complex structures (Ward et al. 1975; Doroquez et al. 2014; Cebul et al. 2020) (Figure 1A). These structures concentrate oxygen-sensing signaling molecules (Gross et al. 2014; Mclachlan et al. 2018) suggesting that similar to cilia, these structures are specialized for sensory functions. Microtubule growth events similar to those observed in ciliated sensory neurons were also reported at the distal dendritic regions of URX, implying the presence of a microtubule organizer such as a remodeled basal body (Harterink et al. 2018). Moreover, a subset of ciliary genes is expressed in URX (Kunitomo et al. 2005; Harterink et al. 2018; Mclachlan et al. 2018). We tested the hypothesis that URX dendrites contain cilia at their distal ends.

L'Hernault SW et al. (1995) International C. elegans Meeting "HOW DO SPERM FIND EGGS AND KNOW WHAT TO DO WHEN THEY FIND THEM?"

Mercer KB, L'Hernault SW

[International C. elegans Meeting, 1995]

Unlike many other organisms, fertilization in C. elegans is an extraordinarily efficient process; normally every sperm produced by a hermaphrodite successfully fertilizes an egg within the spermatheca (Ward, S. and J. Carrel, 1979 Dev. Biol. 73: 304). Many mutants that affect spermatogenesis and, consequently, disrupt fertilization have been selected by both ours and the Ward laboratory. Most of these mutants have obvious cytological abnormalities, so their defects in fertilization are perhaps not surprising. We have been studying mutants that complete spermatogenesis and form spermatozoa with superficially normal cytology but are defective in hermaphrodite self-fertilization. So far, our studies have been confined to mutants that map to three genes: spe-9, spe-13 and spe-16. Mutant males were assessed for sperm transfer by using spe-8 hermaphrodites as recipients; spe-8 hermaphrodites are self-sterile except when they receive "competent" seminal fluid from a male, which allows self-fertility (Shakes, D. & Ward, S. 1989 Dev. Biol. 134: 189). In such tests, none of the examined mutant males had defects in seminal fluid transfer to hermaphrodites. However, spe- 13 or spe-16 males cannot sire progeny in cross- fertilization tests while spe-9 males (either of two ts alleles) can sire cross progeny. Two recently recovered nonconditional spe-9 mutants are being examined to determine if stronger loss of function mutants also exhibit this phenotype. Double mutants that are spe-9ts and spe- 13ts are nonconditionally sterile, suggesting some form of synergism between these two mutations. We are in the process of examining spe-13 and spe-16 in order to determine if male derived spermatozoa can target properly to the hermaphrodite spermatheca, even though they cannot fertilize eggs.

Starr, D. (2015) International Worm Meeting "Cloning the male sterile, sperm defective genes fer-2 and fer-3."

Starr, D.

[International Worm Meeting, 2015]

We all know that the sperm of our favorite model organism are weird. But, did you know they do not have a membranous nuclear envelope? Wolf et al (1978) showed using EM that sperm nuclei are surrounded by a diffuse perinuclear halo that encapsulates the condensed chromatin and the centriole. The peri-nuclear halo is hypothesized to consist of RNA and protein. Sam Ward and his lab performed forward genetic screens for fertilization-defective mutants that disrupted spermatogenesis (Argon and Ward 198). Their ultrastructural analyses identified an interesting disruption of the perinuclear halo in fer-2, fer-3, or fer-4 mutant sperm. The mutant sperm lack the perinuclear halo. Instead, tubular complexes ~50 nm in diameter seem to originate from near the nucleus and radiate away from the nucleus (Ward et al 1981). Given my long-term interest in the nuclear envelope, I set out to clone these fer mutants with the goal of identifying components of the perinuclear halo as an alternative to the nuclear envelope that normally circles eukaryotic nuclei. Using whole-genome sequencing, I have identified possible molecular lesions for fer-2 and fer-3. For fer-2, I identified a nonsense mutation near the beginning of the mib-1 gene. RNAseq and microarray experiments deposited in Wormbase show that mib-1 expression is enriched in sperm. The MIB-1 protein is a predicted RING Zn-finger E3 Ubiquitin ligase with ankyrin repeats that is conserved with fly and vertebrate Mindbomb. fer-3 had a nonsense mutation in the eri-3 gene. Mutations in eri-3 were previously shown to cause temperature-sensitive male sterility. ERI-3 is a nematode-specific accessory component of DICER.

Takashi Murayama et al. (2006) East Asia C. elegans Meeting "Chemotactic response toward basic pH in C. elegans"

Ichiro Maruyama, Takashi Murayama

[East Asia C. elegans Meeting, 2006]

C. elegans responds to water-soluble chemicals such as ions and amino acids. We are especially interested in the response to OH^- as basic pH. In C. elegans, acid pH (H⁺) is sensed as a nociceptive stimuli, and basic pH (OH^-) is sensed as an attractive stimuli (Ward, 1973; Dusenbery, 1974). Sambongi et al. (2000) have shown some of amphid chemosensory neurons are responsible for acid pH avoidance by ablating these neurons. In general, however, a neural mechanism underlying OH^- response in any animals still remains to be explored. To investigate the OH^- chemosensation, we have developed an assay system. On the assay plate with a pH gradient from pH 7.0 to pH 10.0, wild-type animals move toward basic pH along the gradient. When some chemotaxis mutants were analyzed with this assay, osm-6, a sensory cilium-defective mutant, did not show any response to basic pH. In addition, some chemotaxis mutants have normal sensory cilia are found to be insensitive to the pH gradient. Now, we are analyzing some other chemotaxis mutants, and trying to isolate mutants showing irregular response to basic pH. Ward (1973), PNAS 70, 817. Dusenbery (1974), J. Exp. Zool. 188, 41. Sambongi et al. (2000), Neuroreport 11, 2229.

Minniti AN et al. (1995) International C. elegans Meeting "THE HERMAPHRODITE-INITIATED SPERMIOGENESIS PATHWAY."

Minniti AN, Nance JF, Sadler C, Ward S

[International C. elegans Meeting, 1995]

During spermiogenesis, round nonmotile spermatids are rapidly transformed into asymmetrical crawling spermatozoa. In addition to spe-8 and spe-12, we found two new genes, spe-27 and spe-29, that when mutated, disrupt spermiogenesis in an identical manner: mutant hermaphrodites are self-sterile, while mutant males are fertile. Sperm from both sexes activate abnormally in vitro, suggesting that these gene products are expressed in both sperm, but only hermaphrodites require them for activation. The hermaphrodite's spermatids, however, can be activated by mating. This phenotype has been explained by a model that has two distinct pathways of activation, one for males and one for hermaphrodites (Shakes and Ward, 1989). The four genes are needed only for the hermaphrodite pathway. spe-27 and spe-12 have been cloned. Their mRNA is found exclusively in the germline during spermatogenesis. Their predicted protein sequences show no similarities to known proteins. We are currently cloning spe-29. To further understand how these genes act during spermiogenesis we are determining the subcellular localization of their gene products. In addition, suppressor analysis is under way (see abstract by Paul Muhlrad and Samuel Ward) to look for other genes in this pathway and for interactions between these genes.