Fan PC et al. (1985) Southeast Asian J Trop Med Public Health "Experimental infection of subperiodic Brugia malayi in laboratory rats with emphasis on evaluation by four techniques."

Hsu YP, Huang CM, Wu CC, Ho CM, Fan PC

[Southeast Asian J Trop Med Public Health, 1985]

Infective larvae of subperiodic B. malayi from South Kalimantan (Borneo), Indonesia collected from laboratory-raised Ae. togoi mosquitoes after feeding on infected mongolian gerbils (Meriones unguiculatus) were inoculated subcutaneously into the groin areas of 15 SD and 36 LE rats. Blood was examined weekly by membrane filtration and thick smears starting 10 weeks post-infection. Microfilariae were found in 3 SD and 4 LE rats, the mf infection rate of 20% and 11% respectively. The prepatent period was significantly shorter in the SD rats (99-112 days) than those in the LE rats (110-153 days). The patent period was longer in the LE rats (208-703 days) than in the SD rats (236-543 days), and the mf density was similar (17.5 mf/20 c.mm blood against 16 mf/20 c.mm blood). At necropsy, 6 (3 female and 3 male) adult worms were recovered from 3 of 6 SD rats and 12 (9 female and 3 male) adult worms from 4 of 20 LE rats; all worms were found in the testes. The results of xenodiagnostic, histochemical staining and measuring spicules and protuberances, demonstrated clearly the difference between both species of Brugia. All dissected Ar. subalbatus mosquitoes exposed to B. pahangi became infected (100%), but none of those to subperiodic B. malayi were infected (0%). The mf of both species of Brugia in thick films stained with naphthol-AS-TR-phosphate showed that the excretory and anal pores of subperiodic B. malayi mf exhibited acid phosphatase activity and only a little activity was seen in other parts; while B. pahangi mf showed heavy diffuse acid phosphatase activity along the entire length of the body.(ABSTRACT TRUNCATED AT 250 WORDS)

Snodgrass, Casey et al. (2013) International Worm Meeting "Conserved ion and amino acid transporters identified as phosphorylcholine modified N-glycoproteins by metabolic labeling with propargylcholine in Caenorhabditis elegans."

Meteer, John, Berninsone, Patricia M., Snodgrass, Casey, Burnham-Marusich, Amanda

[International Worm Meeting, 2013]

Phosphorylcholine (Pc) modification of proteins by pathogens has been implicated in mediating host-pathogen interactions. In parasitic nematodes, Pc modulates the host's immune response to favor nematode survival. Caenorhabditis elegans expresses Pc-modified N-linked glycans, offering an attractive non-parasitic model to study the biology of Pc-modification, but a lack of selective purification tools limits the number of known Pc targets. Here we show that Pc-modified N-glycoproteins can be identified in C. elegans embryonic cells by metabolic labeling with propargylcholine, an alkyne-modified choline analog. Cu(I)-catalyzed cycloaddition with biotin-azide enables streptavidin purification, high-throughput liquid chromatography and mass spectrometry identification of propargyl-labeled proteins. We report 21 novel Pc-modified N-glycoproteins and their sites of Pc-N-glycan attachment. Ion and amino acid transporters as well as synaptic proteins are among those identified as Pc-modified, suggesting a function for Pc beyond immunomodulation. This work provides a method to study Pc-modified proteins in C. elegans and related nematodes.

Bunte, Myrna et al. (2021) International Worm Meeting "Unravelling the identity of phosphorylcholine-transferring enzymes in C. elegans"

Riksen, Joost, Patrian, Marta, Garcia Perez, Elena, Wilbers, Ruud, Kammenga, Jan, Bunte, Myrna, Schots, Arjen

[International Worm Meeting, 2021]

The attachment of phosphorylcholine on carbohydrates (PC-glycans) is a common modification for nematodes including C. elegans. The presence of PC appears to be important for the development of nematodes and for immunomodulation by parasitic nematodes. PC-glycoprotein ES-62 of Acanthoceilonema vitae directs immune cells towards an anti-inflammatory phenotype and this immunomodulatory property appears to be dependent on the presence of PC-glycans. This makes glycoproteins with PC-glycans interesting therapeutic agents to combat immune disorders such as rheumatoid arthritis, systemic lupus erythematosus and asthma. However, the biosynthetic pathway of PC-glycans is not completely understood, where especially the identity of the PC-transferring enzyme, or PC-transferase, is not fully elucidated. There are strong indications suggesting that fukutin-related genes potentially encode for this PC-transferase. In this study, we examined whether four selected fukutin-related genes of C. elegans (W02B3.4, T07A5.1, T07D3.4 and Y22D7AL.11) are involved in the biosynthetic pathway of PC glycans. With CRISPR/Cas9 technology we created C. elegans knock-out lines for each fukutin-related gene, but no significant reduction of PC was observed. Interestingly, one mutant line of W02B3.4 showed an increase of PC due to a potentially introduced signal peptide, indicating that the W02B3.4 gene could encode for a PC-transferase. Currently, we are combining mutant lines into double/triple/quadruple knock-out lines, which may provide more insight whether these fukutin-related genes encode for PC-transferases. These findings will contribute to our understanding of the pathway for PC-glycan biosynthesis, offering potential opportunities for design and synthesis of PC-glycan therapeutics.

Snodgrass CJ et al. (2015) Glycobiology "Conserved ion and amino acid transporters identified as phosphorylcholine-modified N-glycoproteins by metabolic labeling with propargylcholine in Caenorhabditis elegans cells."

Burnham-Marusich AR, Meteer JC, Berninsone PM, Snodgrass CJ

[Glycobiology, 2015]

Phosphorylcholine (PC) modification of proteins by pathogens has been implicated in mediating host-pathogen interactions. Parasitic nematodes synthesize PC-modified biomolecules that can modulate the host's antibody and cytokine production to favor nematode survival, contributing to long-term infections. Only two nematode PC-modified proteins (PC-proteins) have been unequivocally identified, yet discovering the protein targets of PC modification will be paramount to understanding the role(s) that this epitope plays in nematode biology. A major hurdle in the field has been the lack of techniques for selective purification of PC-proteins. The nonparasitic nematode Caenorhabditis elegans expresses PC-modified N-linked glycans, offering an attractive model to study the biology of PC-modification. We developed a robust method to identify PC-proteins by metabolic labeling of primary embryonic C. elegans cells with propargylcholine, an alkyne-modified choline analog. Cu(I)-catalyzed cycloaddition with biotin-azide enables streptavidin purification and subsequent high-throughput LC-MS identification of propargyl-labeled proteins. All proteins identified using stringent criteria are known or predicted to be membrane or secreted proteins, consistent with the model of a Golgi-resident, putative PC-transferase. Of the 55 PC-N-glycosylation sites reported, 33 have been previously observed as N-glycosylation sites in high-throughput screens of C. elegans. Several identified PC-proteins are nematode-specific proteins, but 10 of the PC-proteins are widely conserved ion transporters and amino acid transporters, while eight are conserved proteins involved in synaptic function. This finding suggests a functional role for PC-modification beyond immunomodulation. The approach presented in this study provides a method to identify PC-proteins in C. elegans and related nematodes.

Sunagawa T et al. (2011) Planta Med "Procyanidins from apples (Malus pumila Mill.) extend the lifespan of Caenorhabditis elegans."

Kanda T, Sami M, Sunagawa T, Shimizu T, Tagashira M, Shirasawa T

[Planta Med, 2011]

Apple polyphenols (AP) mainly consist of procyanidins (PC), which are composed of (-)-epicatechins and (+)-catechins. In order to investigate the antiageing effects of PC, we measured the lifespan of CAENORHABDITIS ELEGANS worms treated with PC. Treatment with 65 g/mL PC extended the mean lifespan of wild-type N2 and FEM-1 worms by 12.1 % and 8.4 %, respectively, i.e., to a similar extent as resveratrol. In addition, treatment with 100 g/mL AP also significantly prolonged the mean lifespan of the same worms by 12.0 % and 5.3 %, respectively, i.e., to a similar extent as PC. In contrast, treatment with (-)-epicatechin did not extend the lifespan of the worms. PC did not modify the growth, food intake, or fecundity of C. elegans. Treatment with PC did not extend the lifespan of MEV-1 worms, which show excessive oxidative stress, indicating that PC had no antioxidant ability in the MEV-1 mutant. Moreover, treatment with PC had no effect on the longevity of SIR-2.1 worms, which lack the activity of SIR-2, a member of the sirtuin family of NAD (+)-dependent protein deacetylases. These results indicated that PC has SIR-2.1-dependent antiageing effects on C. elegans.

Sonani RR et al. (2017) Bioresour Technol "Purification and antioxidant activity of phycocyanin from Synechococcus sp. R42DM isolated from industrially polluted site."

Singh NK, Jakharia K, Madamwar D, Sonani RR, Chaubey MG, Bhastana B, Patel S

[Bioresour Technol, 2017]

The cyanobacterium Synechococcus sp. R42DM, isolated from an industrially polluted site Vatva, Gujarat, India was recognized to produce phycocyanin (PC) as major phycobiliprotein. In present study, the combinatorial approach of chemical and physical methods i.e. Triton-X 100 treatment and ultra-sonication was designed for extraction of PC. From cell extract, the intact and functional-PC was purified up to purity 4.03 by ammonium sulphate fractionation and ion-exchange chromatography. The PC displayed considerable in vitro antioxidant and radical-scavenging activity. This PC was further noticed to scavenge intracellular-ROS and to increase tolerance against thermal and oxidative stress in Caenorhabditis elegans. Moreover, the PC was noticed to improve the physiological behaviour and longevity of C. elegans. In addition, the PC showed remarkable stability under physico-chemical stressors, which is desirable for their use in biomedical applications. In conclusion, present paper added up evidence in support of the prospective use of PC as an antioxidant nutraceutical.

Snodgrass, Casey et al. (2015) International Worm Meeting "Phosphorylcholine-modified glycoproteins involved in host immune modulation by parasitic nematodes: a chemical biology approach developed in C. elegans ."

Burnham-Marusich, Amanda, Meteer, John, Snodgrass, Casey, Berninsone, Patricia

[International Worm Meeting, 2015]

Parasitic nematodes synthesize phosphorylcholine (PC)-modified biomolecules that can modulate the host's antibody and cytokine production to favor nematode survival, contributing to long-term infections. While discovering the protein targets of PC modification is critical to understand the role(s) that this epitope plays in host immune modulation, only two nematode PC-modified proteins (PC-proteins) had been previously identified. The non-parasitic nematode Caenorhabditis elegans expresses PC-modified N-linked glycans, offering an attractive model to study the biology of PC-modification. Using C. elegans as model system, we developed a robust method to identify PC-modified proteins by metabolic labeling primary embryonic cells with propargylcholine, an alkyne-modified choline analog. Cu(I)-catalyzed cycloaddition with biotin-azide enables streptavidin purification and subsequent high-throughput liquid chromatography and mass spectrometry identification of propargyl-labeled proteins. All C. elegans proteins identified using stringent criteria are known or predicted to be membrane or secreted proteins, consistent with the model of a Golgi-resident, putative PC-transferase. Of the 55 PC-N-glycosylation sites reported, 33 have been previously observed as N-glycosylation sites in high-throughput screens of C. elegans. This sensitive and specific method will enable the comprehensive identification of PC-modified glycoproteins in parasitic nematodes and aid in our understanding of the function of PC-modified targets in modulation of the host immune system. .

Jenson, Kevin J et al. (2013) International Worm Meeting "Using C. elegans to identify the nematode phosphorycholine transferase."

Jenson, Kevin J, Berninsone, Patricia M

[International Worm Meeting, 2013]

Parasitic nematodes afflict an estimated 120 million humans worldwide, and at least one billion more are at risk of infection in endemic areas. Available chemotherapeutics target only distinct larval stages and show severe side effects. Furthermore, reported resistance to certain anthelminthics indicates the need for development of new drugs. Nematode-specific biomolecules and the enzymes involved in their biosynthesis provide a rich field for drug target discovery. Phosphorylcholine (PC), a small haptenic molecule, is found linked to glycans in many parasitic nematodes, but not in mammals. PC modification of proteins by pathogens has been implicated in mediating host-pathogen interactions through immunomodulation. This is done by inducing a Th2-type anti-inflammatory response while simultaneously inhibiting T- and B- cell production. Thus, immune attention is diverted away from fighting the parasite. Because phosphorylcholination is not utilized by mammalian cells, the PC transferase catalyzing this reaction makes an attractive anthelminthic drug target. A major barrier to exploring the PC modification as a drug target is that no protein catalyzing the PC transferase reaction has been identified in eukaryotes. The free-living nematode C. elegans synthesizes PC-modified glycans and thus offers an attractive model to investigate the biology of PC-modified molecules. PC-modification in nematodes occurs when a putative transferase resident to the Golgi utilizes phosphatidylcholine as a PC donor to decorate terminal N-acetylglucosamine (GlcNAc) residues of N-linked glycoproteins. However, the enzyme catalyzing the addition of PC to N-linked glycoproteins remains unknown. The goal of this project is to identify the phosphorylcholine transferase in C. elegans. We have performed bioinformatic analysis and found that eight C. elegans genes have homology to key domains of known bacterial PC transferases. We have also compiled a list of Golgi-localized proteins that have no known function. It is our hypothesis that the PC transferase lies within these lists.

Nabeshima K (2012) Worm "Collaborative homologous pairing during C. elegans meiosis."

Nabeshima K

[Worm, 2012]

In preparation for meiotic chromosome segregation, homologous chromosomes need to pair, synapse (i.e., assemble the synaptonemal complex, SC), and then recombine to generate a physical linkage (i.e., chiasma) between them. In many organisms meiotic pairing capacity distributed along the entire chromosome length supports presynaptic alignment. In contrast, the prevailing model for C. elegans proposes that presynaptic homologous pairing is performed solely by a master pairing-site, the pairing center (PC). In this model, the remaining chromosomal regions (the non-PC regions) are not actively involved in presynaptic pairing, and the SC assembling from the PC aligns the homologous chromosomes along non-PC regions and holds them together. Our recent work, however, demonstrates that C. elegans chromosomes establish presynaptic alignment along the entire chromosome length, suggesting that the non-PC regions are also actively involved in the presynaptic pairing process. Furthermore, we have also discovered that the chromodomain protein MRG-1 facilitates this presynaptic non-PC pairing. The phenotype of the mrg-1 mutant indicates that the PC and the non-PC collaborate in successful pairing and synapsis. Therefore, homologous pairing mechanisms in C. elegans possibly share more similarity with those in other organisms than previously thought. Here, we elaborate on these observations and discuss a hypothetical model for presynaptic pairing in C. elegans based on our novel findings.

Zhu J et al. (2023) J Genet Genomics "Phosphatidylcholine deficiency increases ferroptosis susceptibility in the C. elegans germline."

Huang X, Zhu J, Shui G, Man Lam S, Meng W

[J Genet Genomics, 2023]

Ferroptosis, a regulated and iron-dependent form of cell death characterized by peroxidation of membrane phospholipids, has tremendous potential for the therapy of human diseases. The causal link between phospholipid homeostasis and ferroptosis is incompletely understood. Here, we reveal that spin-4, a previously identified regulator of the "B12-one-carbon cycle-phosphatidylcholine (PC)" pathway, sustains germline development and fertility by ensuring PC sufficiency in the nematode Caenorhabditis elegans. Mechanistically, SPIN-4 regulates lysosomal activity which is required for B12-associated PC synthesis. PC deficiency-induced sterility can be rescued by reducing the levels of polyunsaturated fatty acids (PUFAs), reactive oxygen species (ROS) , and redox-active iron, which indicates that the sterility is mediated by germline ferroptosis. These results highlight the critical role of PC homeostasis in ferroptosis susceptibility and offer a new target for pharmacological approaches.