Aminopeptidases inCaenorhabditis elegansandPanagrellus redivivus: detection using peptide and non-peptide substrates

2002 ◽  
Vol 76 (1) ◽  
pp. 45-52 ◽  
Author(s):  
E.P. Masler
Keyword(s):  
Caenorhabditis Elegans ◽  
Substance P ◽  
Bioactive Peptides ◽  
Specific Activity ◽  
Adipokinetic Hormone ◽  
Free Living ◽  
Peptide Substrates ◽  
C Elegans ◽  
Panagrellus Redivivus ◽  
Chromatographic Profiles

AbstractAminopeptidase activities were detected in extracts of the free-living nematodesCaenorhabditis elegansandPanagrellus redivivususing the aminoacyl substrate L-alanine-4-nitroanilide. The activities exhibited similarities in Km (C.elegans= 2.22 mM;P.REDIVIVUS= 2.09 Mm) and specific activity (C.elegans=1.38±0.43 mAU min-1 μg-1;P. redivivus, 1.23±0.18 mAU min-1 μg-1). Each is inhibited competitively by amastatin (C. elegansIC50=0.46 μm;P. redivivusIC50=15.90 μm) and non-competitively by leuhistin (C. elegansIC50=3.00 μm;P. redivivusIC50=37.35 μm). The bioactive peptides adipokinetic hormone and substance P decrease the apparent aminopeptidase activities of each extract suggesting that the peptides compete with the Ala-pNA as substrates. With each extract, adipokinetic hormone appeared to be the more effective substrate. Digestion of adipokinetic hormone byC. elegansandP. redivivusextracts in the presence and absence of 1 mm amastatin produced distinct chromatographic profiles that suggest different digestion patterns for the two species. However, amastatin had clear effects on chromatographic profiles from each species indicating that an aminopeptidase is involved in the digestion of the peptide substrates. The data presented indicate that extracts of free-living nematodes are capable of metabolizing peptide hormones, and that this metabolism involves substrate-selective aminopeptidases.

Transfer and tissue-specific accumulation of components in embryos of Caenorhabditis elegans and dolichura: in vivo analysis with a low-cost signal

Development ◽  
1992 ◽  
Vol 114 (2) ◽  
pp. 317-330 ◽  
Author(s):  
O. Bossinger ◽  
E. Schierenberg
Keyword(s):  
Caenorhabditis Elegans ◽  
Low Cost ◽  
Free Living ◽  
Tissue Specific ◽  
C Elegans ◽  
Specific Accumulation ◽  
In Vivo Analysis

The pattern of autofluorescence in the two free-living namatodes Rhabditis dolichura and Caenorhabditis compared. In C. elegans, during later embryogenesis cells develop a typical bluish autofluorescence as illumination, while in Rh. dolichura a strong already present in the unfertilized egg. Using a new,

scholarly journals Haematophagic Caenorhabditis elegans

Parasitology ◽  
2018 ◽  
Vol 146 (3) ◽  
pp. 314-320 ◽  
Author(s):  
Veeren M Chauhan ◽  
David I Pritchard
Keyword(s):  
Caenorhabditis Elegans ◽  
Neutralizing Antibodies ◽  
Fluorescent Microscopy ◽  
Free Living ◽  
C Elegans ◽  
Vaccine Responses ◽  
Nematode Parasites ◽  
Significant Difference ◽  
Free Living Nematode ◽  
Bright Emission

AbstractCaenorhabditis elegans is a free-living nematode that resides in soil and typically feeds on bacteria. We postulate that haematophagic C. elegans could provide a model to evaluate vaccine responses to intestinal proteins from hematophagous nematode parasites, such as Necator americanus. Human erythrocytes, fluorescently labelled with tetramethylrhodamine succinimidyl ester, demonstrated a stable bright emission and facilitated visualization of feeding events with fluorescent microscopy. C. elegans were observed feeding on erythrocytes and were shown to rupture red blood cells upon capture to release and ingest their contents. In addition, C. elegans survived equally on a diet of erythrocytes. There was no statistically significant difference in survival when compared with a diet of Escherichia coli OP50. The enzymes responsible for the digestion and detoxification of haem and haemoglobin, which are key components of the hookworm vaccine, were found in the C. elegans intestine. These findings support our postulate that free-living nematodes could provide a model for the assessment of neutralizing antibodies to current and future hematophagous parasite vaccine candidates.

Many transcribed regions of the Onchocerca volvulus genome contain the spliced leader sequence of Caenorhabditis elegans

1990 ◽  
Vol 10 (6) ◽  
pp. 2765-2773
Author(s):  
W L Zeng ◽  
C M Alarcon ◽  
J E Donelson
Keyword(s):  
Caenorhabditis Elegans ◽  
Repeat Unit ◽  
5S Rrna ◽  
Onchocerca Volvulus ◽  
Rrna Genes ◽  
Free Living ◽  
Spliced Leader ◽  
C Elegans ◽  
Perfect Sequence ◽  
5S Rrna Genes

Genomic DNAs of the related parasitic nematodes Onchocerca volvulus and Dirofilariae immitis, and a cDNA library of O. volvulus, were examined for the presence of the 22-nucleotide spliced leader (SL) found at the 5' ends of 10 to 15% of the mRNAs in the free-living nematode Caenorhabditis elegans. As in C. elegans, genes for the SL RNA are linked to the repetitive 5S rRNA genes of O. volvulus and D. immitis, but unlike C. elegans, they are in the same orientation as the 5S rRNA genes within the repeat unit. In O. volvulus the SL sequence is also encoded at more than 30 additional genomic locations and occurs at interior sites within many transcripts. Sequence determinations of four different cDNAs of O. volvulus, each containing an internal copy of the SL within a conserved 25mer, and one corresponding genomic DNA clone indicate that this sequence is not trans spliced onto these RNAs, but is encoded within the genes. The RNAs of two of these cDNAs appear to be developmentally regulated, since they occur in adult O. volvulus but were not detected in the infective L3 stage larvae. In contrast, actin mRNAs are present at all developmental stages, and at least one actin mRNA species contains a trans-spliced 5' SL. The internal locations of the SL in various transcripts and its perfect sequence conservation among parasitic and free-living nematodes argues that it serves specific, and perhaps multiple, functions for these organisms.

Effect of structurally related flavonoids from Zuccagnia punctata Cav. on Caenorhabditis elegans

2014 ◽  
Vol 60 (1) ◽  
Author(s):  
Romina E. D’Almeida ◽  
María R. Alberto ◽  
Phillip Morgan ◽  
Margaret Sedensky ◽  
María I. Isla
Keyword(s):  
Caenorhabditis Elegans ◽  
Larval Development ◽  
Free Living ◽  
Egg Hatching ◽  
C Elegans ◽  
Therapeutic Drugs ◽  
Aerial Parts ◽  
Free Living Nematode ◽  
Development Processes ◽  
Anthelmintic Effect

AbstractZuccagnia punctata Cav. (Fabaceae), commonly called jarilla macho or pus-pus, is being used in traditional medicine as an antiseptic, anti-inflammatory and to relieve muscle and bone pain. The aim of this work was to study the anthelmintic effects of three structurally related flavonoids present in aerial parts of Z. punctata Cav. The biological activity of the flavonoids 7-hydroxyflavanone (HF), 3,7-dihydroxyflavone (DHF) and 2´,4´-dihydroxychalcone (DHC) was examined in the free-living nematode Caenorhabditis elegans. Our results showed that among the assayed flavonoids, only DHC showed an anthelmintic effect and alteration of egg hatching and larval development processes in C. elegans. DHC was able to kill 50% of adult nematodes at a concentration of 17 μg/mL. The effect on larval development was observed after 48 h in the presence of 25 and 50 μg/mL DHC, where 33.4 and 73.4% of nematodes remained in the L3 stage or younger. New therapeutic drugs with good efficacy against drug-resistant nematodes are urgently needed. Therefore, DHC, a natural compound present in Z. punctata, is proposed as a potential anthelmintic drug.

Nutritional requirements for pantothenate, pantethine or coenzyme A in the free-living nematode Caenorhabditis elegans

Nematology ◽  
2005 ◽  
Vol 7 (5) ◽  
pp. 761-766 ◽  
Author(s):  
Nancy Lu ◽  
Rekha Balachandar
Keyword(s):  
Caenorhabditis Elegans ◽  
Coenzyme A ◽  
Active Form ◽  
Calcium Pantothenate ◽  
Axenic Medium ◽  
Nematode Caenorhabditis Elegans ◽  
Free Living ◽  
C Elegans ◽  
Free Living Nematode ◽  
Mild Toxicity

AbstractCaenorhabditis elegans is a free-living nematode cultured in an axenic medium, the Caenorhabditis elegans Maintenance Medium (CeMM), which contains B-vitamins, salts, amino acids, nucleic acid substituents, growth factors and glucose as an energy source. After initial experiments established that either pantothenate or pantethine would satisfy the vitamin B5 requirement in C. elegans, reproduction in the nematodes was measured in eight equimolar concentrations of calcium pantothenate, pantethine or coenzyme A. The optimal levels for pantothenate were found to be 7.5, 30 and 120 μg ml−1. The optimal levels for pantethine and coenzyme A were found to be 35 μg ml−1 and 100 μg ml−1, respectively. Among the three compounds, coenzyme A (at 100 μg/ml) supported a significantly greater population growth and, perhaps, is a more metabolically active form. Mild toxicity was demonstrated for pantothenate at 480μg ml−1, pantethine at 560 and 140 μg ml−1, and coenzyme A was found to exhibit toxicity at 410 and 1700 μg ml−1. Based on our results, we recommend that in the future the CeMM could be supplemented with pantothenate (7.5 μg ml−1) alone.

scholarly journals An Enterotoxin-Like Binary Protein from Pseudomonas protegens with Potent Nematicidal Activity

2017 ◽  
Vol 83 (19) ◽  
Author(s):  
Jun-Zhi Wei ◽  
Daniel L. Siehl ◽  
Zhenglin Hou ◽  
Barbara Rosen ◽  
Jarred Oral ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Caenorhabditis Elegans ◽  
Cholera Toxin ◽  
Nematicidal Activity ◽  
Binary Toxin ◽  
Parasitic Nematodes ◽  
Free Living ◽  
Subunit A ◽  
Content Type ◽  
Panagrellus Redivivus

ABSTRACT Soil microbes are a major food source for free-living soil nematodes. It is known that certain soil bacteria have evolved systems to combat predation. We identified the nematode-antagonistic Pseudomonas protegens strain 15G2 from screening of microbes. Through protein purification we identified a binary protein, designated Pp-ANP, which is responsible for the nematicidal activity. This binary protein inhibits Caenorhabditis elegans growth and development by arresting larvae at the L1 stage and killing older-staged worms. The two subunits, Pp-ANP1a and Pp-ANP2a, are active when reconstituted from separate expression in Escherichia coli. The binary toxin also shows strong nematicidal activity against three other free-living nematodes (Pristionchus pacificus, Panagrellus redivivus, and Acrobeloides sp.), but we did not find any activity against insects and fungi under test conditions, indicating specificity for nematodes. Pp-ANP1a has no significant identity to any known proteins, while Pp-ANP2a shows ∼30% identity to E. coli heat-labile enterotoxin (LT) subunit A and cholera toxin (CT) subunit A. Protein modeling indicates that Pp-ANP2a is structurally similar to CT/LT and likely acts as an ADP-ribosyltransferase. Despite the similarity, Pp-ANP shows several characteristics distinct from CT/LT toxins. Our results indicate that Pp-ANP is a new enterotoxin-like binary toxin with potent and specific activity to nematodes. The potency and specificity of Pp-ANP suggest applications in controlling parasitic nematodes and open an avenue for further research on its mechanism of action and role in bacterium-nematode interaction. IMPORTANCE This study reports the discovery of a new enterotoxin-like binary protein, Pp-ANP, from a Pseudomonas protegens strain. Pp-ANP shows strong nematicidal activity against Caenorhabditis elegans larvae and older-staged worms. It also shows strong activity on other free-living nematodes (Pristionchus pacificus, Panagrellus redivivus, and Acrobeloides sp.). The two subunits, Pp-ANP1a and Pp-ANP2a, can be expressed separately and reconstituted to form the active complex. Pp-ANP shows some distinct characteristics compared with other toxins, including Escherichia coli enterotoxin and cholera toxin. The present study indicates that Pp-ANP is a novel binary toxin and that it has potential applications in controlling parasitic nematodes and in studying toxin-host interaction.

The use of Caenorhabditis elegans in parasitic nematode research

Parasitology ◽  
2004 ◽  
Vol 128 (S1) ◽  
pp. S49-S70 ◽  
Author(s):  
J. S. GILLEARD
Keyword(s):  
Caenorhabditis Elegans ◽  
Parasitic Nematode ◽  
Expression System ◽  
Nematode Species ◽  
Current Status ◽  
Evolutionary Development ◽  
Parasitic Nematodes ◽  
Free Living ◽  
C Elegans ◽  
Free Living Nematode

There is increasing interest in the use of the free-living nematode Caenorhabditis elegans as a tool for parasitic nematode research and there are now a number of compelling examples of its successful application. C. elegans has the potential to become a standard tool for molecular helminthology researchers, just as yeast is routinely used by molecular biologists to study vertebrate biology. However, in order to exploit C. elegans in a meaningful manner, we need a detailed understanding of the extent to which different aspects of C. elegans biology have been conserved with particular groups of parasitic nematodes. This review first considers the current state of knowledge regarding the conservation of genome organisation across the nematode phylum and then discusses some recent evolutionary development studies in free-living nematodes. The aim is to provide some important concepts that are relevant to the extrapolation of information from C. elegans to parasitic nematodes and also to the interpretation of experiments that use C. elegans as a surrogate expression system. In general, examples have been specifically chosen because they highlight the importance of careful experimentation and interpretation of data. Consequently, the focus is on the differences that have been found between nematode species rather than the similarities. Finally, there is a detailed discussion of the current status of C. elegans as a heterologous expression system to study parasite gene function and regulation using successful examples from the literature.

scholarly journals Caenorhabditis elegans Infrared-Based Motility Assay Identified New Hits for Nematicide Drug Development

2019 ◽  
Vol 6 (1) ◽  
pp. 29 ◽  
Author(s):  
Gastón Risi ◽  
Elena Aguilera ◽  
Enrique Ladós ◽  
Gonzalo Suárez ◽  
Inés Carrera ◽  
...  
Keyword(s):  
Caenorhabditis Elegans ◽  
Model Organism ◽  
Proof Of Concept ◽  
Free Living ◽  
C Elegans ◽  
Nematode Parasites ◽  
Automated Assay ◽  
Free Living Nematode ◽  
Vertebrate Model ◽  
Electronegative Atom

Nematode parasites have a profound impact on humankind, infecting nearly one-quarter of the world’s population, as well as livestock. There is a pressing need for discovering nematicides due to the spread of resistance to currently used drugs. The free-living nematode Caenorhabditis elegans is a formidable experimentally tractable model organism that offers key advantages in accelerating nematicide discovery. We report the screening of drug-like libraries using an overnight high-throughput C. elegans assay, based on an automated infrared motility reader. As a proof of concept, we screened the “Pathogen Box” library, and identical results to a previous screen using Haemonchus contortus were obtained. We then screened an in-house library containing a diversity of compound families. Most active compounds had a conjugation of an unsaturation with an electronegative atom (N, O, or S) and an aromatic ring. Importantly, we identified symmetric arylidene ketones and aryl hydrazine derivatives as novel nematicides. Furthermore, one of these compounds, (1E,2E)-1,2-bis(thiophen-3-ylmethylene)hydrazine, was active as a nematicide at 25 µm, but innocuous to the vertebrate model zebrafish at 50 µm. Our results identified novel nematicidal scaffolds and illustrate the value of C. elegans in accelerating nematicide discovery using a nonlabor-intensive automated assay that provides a simple overnight readout.

Developmental strategies during early embryogenesis of Caenorhabditis elegans

Development ◽  
1986 ◽  
Vol 97 (Supplement) ◽  
pp. 31-44
Author(s):  
Einhard Schierenberg
Keyword(s):  
Caenorhabditis Elegans ◽  
Parasitic Nematode ◽  
Soil Nematode ◽  
Developmental Studies ◽  
Central Question ◽  
Unique Combination ◽  
Nematode Caenorhabditis Elegans ◽  
Free Living ◽  
C Elegans ◽  
Multicellular Structure

How the complex, multicellular structure of an organism is generated from the information contained in the uncleaved egg is a central question in developmental studies. Nematodes are particularly suitable for studying this question. A unique combination of favourable properties, including transparent eggshell, normal embryogenesis under the microscope outside the mother, small number of cells and rapid, reproducible development made nematodes classic models for developmental biologists (for reviews see Chitwood & Chitwood, 1974; von Ehrenstein & Schierenberg, 1980). In addition to the attractive features mentioned above, the free-living soil nematode Caenorhabditis elegans (Fig. 1) is also well suited for analysis of the genetic control of development (Brenner, 1974) unlike the classically studied parasitic nematode Parascaris equorum (Ascaris megalocephala). Recently cellular (e.g. Sulston, Schierenberg, White & Thomson, 1983) and genetic (e.g. Sternberg & Horvitz, 1984) aspects of development have been studied extensively in C. elegans.

scholarly journals Ingestion of Salmonella enterica Serotype Poona by a Free-Living Nematode, Caenorhabditis elegans, and Protection against Inactivation by Produce Sanitizers

2003 ◽  
Vol 69 (7) ◽  
pp. 4103-4110 ◽  
Author(s):  
Krishaun N. Caldwell ◽  
Barbara B. Adler ◽  
Gary L. Anderson ◽  
Phillip L. Williams ◽  
Larry R. Beuchat
Keyword(s):  
Lactic Acid ◽  
Caenorhabditis Elegans ◽  
Salmonella Enterica ◽  
Fruits And Vegetables ◽  
Sodium Chlorite ◽  
Water Control ◽  
Free Living ◽  
C Elegans ◽  
Significant Difference ◽  
Free Living Nematode

ABSTRACT Free-living nematodes are known to ingest food-borne pathogens and may serve as vectors to contaminate preharvest fruits and vegetables. Caenorhabditis elegans was selected as a model to study the effectiveness of sanitizers in killing Salmonella enterica serotype Poona ingested by free-living nematodes. Aqueous suspensions of adult worms that had fed on S. enterica serotype Poona were treated with produce sanitizers. Treatment with 20 μg of free chlorine/ml significantly (α = 0.05) reduced the population of S. enterica serotype Poona compared to results for treating worms with water (control). However, there was no significant difference in the number of S. enterica serotype Poona cells surviving treatments with 20 to 500 μg of chlorine/ml, suggesting that reductions caused by treatment with 20 μg of chlorine/ml resulted from inactivation of S. enterica serotype Poona on the surface of C. elegans but not cells protected by the worm cuticle after ingestion. Treatment with Sanova (850 or 1,200 μg/ml), an acidified sodium chlorite sanitizer, caused reductions of 5.74 and 6.34 log10 CFU/worm, respectively, compared to reductions from treating worms with water. Treatment with 20 or 40 μg of Tsunami 200/ml, a peroxyacetic acid-based sanitizer, resulted in reductions of 4.83 and 5.34 log10 CFU/worm, respectively, compared to numbers detected on or in worms treated with water. Among the organic acids evaluated at a concentration of 2%, acetic acid was the least effective in killing S. enterica serotype Poona and lactic acid was the most effective. Treatment with up to 500 μg of chlorine/ml, 1% hydrogen peroxide, 2,550 μg of Sanova/ml, 40 μg of Tsunami 200/ml, or 2% acetic, citric, or lactic acid had no effect on the viability or reproductive behavior of C. elegans. Treatments were also applied to cantaloupe rind and lettuce inoculated with S. enterica serotype Poona or C. elegans that had ingested S. enterica serotype Poona. Protection of ingested S. enterica serotype Poona against sanitizers applied to cantaloupe was not evident; however, ingestion afforded protection of the pathogen on lettuce. These results indicate that S. enterica serotype Poona ingested by C. elegans may be protected against treatment with chlorine and other sanitizers, although the basis for this protection remains unclear.

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