Characterization of heat shock protein 70 gene fromHaemonchus contortusand its expression and promoter analysis inCaenorhabditis elegans

Parasitology ◽  
2013 ◽  
Vol 140 (6) ◽  
pp. 683-694 ◽  
Author(s):  
HONGLI ZHANG ◽  
QIANJIN ZHOU ◽  
YI YANG ◽  
XUEQIU CHEN ◽  
BAOLONG YAN ◽  
...  
Keyword(s):  
Heat Shock ◽  
Haemonchus Contortus ◽  
Expression Patterns ◽  
Small Ruminants ◽  
Green Fluorescence Protein ◽  
Bioinformatic Analysis ◽  
Parasitic Nematodes ◽  
Hsp70 Mrna ◽  
C Elegans ◽  
Larval Stages

SUMMARYHaemonchus contortusinfections in small ruminants are of major economic importance worldwide. Heat shock proteins (HSPs) are a family of molecular chaperones that play important roles in the process of invasion and survival of nematodes. Although HSP70 has been identified in several parasitic nematodes, little is known of its distribution and function inHaemonchus contortus. The aims of this study were to characterize HSP70 fromHaemonchus contortus(designed as Hc-hsp70), express Hc-hsp70 and analyse the promoter activity inCaenorhabditis elegans. Bioinformatic analysis revealed that the open reading frame of the Hc-hsp70 cDNA encodes a 646-amino acid peptide, which is highly conserved in comparison to HSP70 in other nematodes. Phylogenetic analysis indicated thatH. contortusis closely related toCaenorhabditis. The 5′-flanking region promoted green fluorescence protein (GFP) expression in the intestine in all larval stages and adult with 2 expression patterns inC. elegans. Expression of Hc-hsp70 mRNA transcripts inC. elegansincreased following 2, 4, 6 h of heat shock and peaked at 4 h. However, its expression induced down-regulation ofhsp-1ofC. elegans. These results suggest that theH. contortushsp70 might have a similar function to that ofC. elegans hsp-1.

Nematode ligand-gated chloride channels: an appraisal of their involvement in macrocyclic lactone resistance and prospects for developing molecular markers

Parasitology ◽  
2007 ◽  
Vol 134 (8) ◽  
pp. 1111-1121 ◽  
Author(s):  
S. McCAVERA ◽  
T. K. WALSH ◽  
A. J. WOLSTENHOLME
Keyword(s):  
Haemonchus Contortus ◽  
Chloride Channels ◽  
Macrocyclic Lactone ◽  
Parasitic Nematodes ◽  
C Elegans ◽  
Molecular Tests ◽  
Cooperia Oncophora ◽  
Macrocyclic Lactone Resistance ◽  
High Level ◽  
Laboratory Models

SUMMARYLigand-gated chloride channels, including the glutamate-(GluCl) and GABA-gated channels, are the targets of the macrocyclic lactone (ML) family of anthelmintics. Changes in the sequence and expression of these channels can cause resistance to the ML in laboratory models, such as Caenorhabditis elegans and Drosophila melanogaster. Mutations in multiple GluCl subunit genes are required for high-level ML resistance in C. elegans, and this can be influenced by additional mutations in gap junction and amphid genes. Parasitic nematodes have a different complement of channel subunit genes from C. elegans, but a few genes, including avr-14, are widely present. A polymorphism in an avr-14 orthologue, which makes the subunit less sensitive to ivermectin and glutamate, has been identified in Cooperia oncophora, and polymorphisms in several subunits have been reported from resistant isolates of Haemonchus contortus. This has led to suggestions that ML resistance may be polygenic. Possible reasons for this, and its consequences for the development of molecular tests for resistance, are explored.

scholarly journals Temporal and spatial expression patterns of the small heat shock (hsp16) genes in transgenic Caenorhabditis elegans.

1992 ◽  
Vol 3 (2) ◽  
pp. 221-233 ◽  
Author(s):  
E G Stringham ◽  
D K Dixon ◽  
D Jones ◽  
E P Candido
Keyword(s):  
Caenorhabditis Elegans ◽  
Heat Shock ◽  
Gene Pair ◽  
Expression Patterns ◽  
Developmentally Regulated ◽  
Noncoding Sequences ◽  
C Elegans ◽  
Gene Pairs ◽  
Temporal And Spatial ◽  
Beta Galactosidase

The expression of the hsp16 gene family in Caenorhabditis elegans has been examined by introducing hsp16-lacZ fusions into the nematode by transformation. Transcription of the hsp16-lacZ transgenes was totally heat-shock dependent and resulted in the rapid synthesis of detectable levels of beta-galactosidase. Although the two hsp16 gene pairs of C. elegans are highly similar within both their coding and noncoding sequences, quantitative and qualitative differences in the spatial pattern of expression between gene pairs were observed. The hsp16-48 promoter was shown to direct greater expression of beta-galactosidase in muscle and hypodermis, whereas the hsp16-41 promoter was more efficient in intestine and pharyngeal tissue. Transgenes that eliminated one promoter from a gene pair were expressed at reduced levels, particularly in postembryonic stages, suggesting that the heat shock elements in the intergenic region of an hsp16 gene pair may act cooperatively to achieve high levels of expression of both genes. Although the hsp16 gene pairs are never constitutively expressed, their heat inducibility is developmentally restricted; they are not heat inducible during gametogenesis or early embryogenesis. The hsp16 genes represent the first fully inducible system in C. elegans to be characterized in detail at the molecular level, and the promoters of these genes should find wide applicability in studies of tissue- and developmentally regulated genes in this experimental organism.

scholarly journals Characterization of a Rhodanese Homologue from Haemonchus Contortus and its Immune-Modulatory Effects on Goat Immune Cells in Vitro

2020 ◽  
Author(s):  
Yujian Wang ◽  
Muhammad Ehsan ◽  
Jianmei Huang ◽  
Kalibixiati Aimulajiang ◽  
RuoFeng Yan ◽  
...  
Keyword(s):  
Haemonchus Contortus ◽  
Mononuclear Cells ◽  
Small Ruminants ◽  
Parasitic Nematodes ◽  
Dependent Manner ◽  
Peripheral Blood Mononuclear ◽  
Nematode Parasites ◽  
Wide Range

Abstract Background: Suppression and modulation of the immune response of the host by nematode parasites have been reported widely. Rhodaneses or thiosulfate: cyanide sulfurtransferases are present in a wide range of organisms, such as archea, bacteria, fungi, plants and animals. Previously, it was reported that a rhodanese homology could bind by goat peripheral blood mononuclear cells (PBMCs) in vivo.Results: In the present study, we cloned and produced recombinant rhodanese protein originated from Haemonchus contortus (rHCRD), which was one of the parasitic nematodes of small ruminants. The effect of this protein on modulating the immunity of goat PBMC and monocyte was studied in the current work. The predominant localization of the natural HCRD protein was verified as the bowel wall and body surface of worms, according to the immunohistochemical tests. It was proved in this study that the serum produced by artificially infecting goats with H. contortus successfully recognized rHCRD which conjugated goat PBMCs. The rHCRD was co-incubated with goat PBMCs to observe the immunomodulatory effect on proliferation, apoptosis and secretion of cytokines exerted by HCRD. The results showed that the interaction of rHCRD suppressed proliferation of goat PBMCs stimulated by ConA but did not induce the apoptosis of goat PBMCs. After rHCRD exposure, the production of TNF-α and IFN-γ were significantly decreased, however, it significantly increased the secretion of IL-10 and TGF-β1 in goat PBMCs. Phagocytotic assay by FITC-dextran internalization showed that rHCRD inhibited the phagocytosis of goat monocytes. Moreover, rHCRD could down-regulate the expression of MHC-II on goat monocytes in a dose-dependent manner. Conclusions: These discoveries proposed a possible target as immunomodulator, which was potentially beneficial to illuminate the interaction between parasites and hosts in the molecular level and hunt for innovative protein species as candidate targets of drug and vaccine.

Constitutive roles for inducible genes: evidence for the alteration in expression of the inducible hsp70 gene in Antarctic notothenioid fishes

2004 ◽  
Vol 287 (2) ◽  
pp. R429-R436 ◽  
Author(s):  
Sean P. Place ◽  
Mackenzie L. Zippay ◽  
Gretchen E. Hofmann
Keyword(s):  
New Zealand ◽  
Heat Shock ◽  
Thermal Stresses ◽  
Expression Patterns ◽  
Antarctic Fish ◽  
Mrna Levels ◽  
Hsp70 Gene ◽  
Hsp70 Mrna ◽  
Inducible Genes ◽  
The Antarctic

Previous research on the Antarctic notothenioid fish Trematomus bernacchii demonstrated the loss of the heat shock response (HSR), a classical cellular defense mechanism against thermal stress, characterized by the rapid synthesis of heat shock proteins (Hsps). In the current study, we examined potential mechanisms for the apparent loss of the HSR in Antarctic notothenioids and, in addition, compared expression patterns of two genes from the 70-kDa Hsp family ( hsc71 and hsp70) in tissues from T. bernacchii to expression patterns in tissues of two closely related temperate notothenioid fishes from New Zealand, Bovichtus variegatus and Notothenia angustata. The results showed that transcript for both the constitutive and inducible genes in the Hsp70 gene family were expressed in detectable levels in all three species. However, only the cold-temperate New Zealand fishes displayed the ability to upregulate the inducible transcript, hsp70. Although hsp70 was present in detectable levels in several tissues of the Antarctic notothen T. bernacchii, in vitro thermal stresses failed to produce a significant increase in mRNA levels. In all species, the expression of the constitutive transcript hsc71 was variable and nonresponsive to temperature increases, even at temperatures as high as 10°C above the ecologically relevant range for the species under study. Field-collected tissues from T. bernacchii (sampled immediately after capture) indicated that hsp70 mRNA was expressed at high levels in field-acclimatized fishes. Thus upregulation of molecular chaperones suggested that low-temperature stress may be significantly denaturing to cellular proteins in Antarctic fish, an observation that was supported by elevated levels of ubiquitin-conjugated protein.

Extending from PARs in Caenorhabditis elegans to homologues in Haemonchus contortus and other parasitic nematodes

Parasitology ◽  
2006 ◽  
Vol 134 (4) ◽  
pp. 461-482 ◽  
Author(s):  
S. NIKOLAOU ◽  
R. B. GASSER
Keyword(s):  
Caenorhabditis Elegans ◽  
Haemonchus Contortus ◽  
Asymmetric Cell Division ◽  
Small Ruminants ◽  
Future Research ◽  
Parasitic Nematodes ◽  
Par Proteins ◽  
Surface Receptors ◽  
Cytoskeletal Dynamics ◽  
Free Living Nematode

Signal transduction molecules play key roles in the regulation of developmental processes, such as morphogenesis, organogenesis and cell differentiation in all organisms. They are organized into ‘pathways’ that represent a coordinated network of cell-surface receptors and intracellular molecules, being involved in sensing environmental stimuli and transducing signals to regulate or modulate cellular processes, such as gene expression and cytoskeletal dynamics. A particularly important group of molecules implicated in the regulation of the cytoskeleton for the establishment and maintenance of cell polarity is the PAR proteins (derived from partition defective in asymmetric cell division). The present article reviews salient aspects of PAR proteins involved in the early embryonic development and morphogenesis of the free-living nematode Caenorhabditis elegans and some other organisms, with an emphasis on the molecule PAR-1. Recent advances in the knowledge and understanding of PAR-1 homologues from the economically important parasitic nematode, Haemonchus contortus, of small ruminants is summarized and discussed in the context of exploring avenues for future research in this area for parasitic nematodes.

scholarly journals Curtisia dentata (Cornaceae) leaf extracts and isolated compounds inhibit motility of parasitic and free-living nematodes

2009 ◽  
Vol 76 (2) ◽  
Author(s):  
L.J. Shai ◽  
E.S. Bizimenyera ◽  
V. Bagla ◽  
L.J. McGaw ◽  
J.N. Eloff
Keyword(s):  
Medicinal Plant ◽  
Betulinic Acid ◽  
Small Ruminants ◽  
Anthelmintic Activity ◽  
Parasitic Nematodes ◽  
Leaf Extracts ◽  
Free Living ◽  
C Elegans

Haemonchus contortus and Trichostrongylus colubriformis are among the most important parasitic nematodes of small ruminants. Caenorhabditis elegans, a free-living nematode, is used as a model for evaluating anthelmintic activity of a variety of test substances. Extracts of several medicinal plants are useful in vitro and in vivo against nematode development. Extracts of Curtisia dentata, a South African medicinal plant, and compounds isolated from leaves of this plant were investigated for anthelmintic activity against T. colubriformis, H. contortus and C. elegans. The acetone and dichloromethane extracts were active against all nematodes at concentrations as low as 160 μg/mℓ. Betulinic acid and lupeol were active against the parasitic nematodes only at the high concentrations of 1 000 and 200 μg/mℓ, respectively. All compounds were effective against C. elegans with active concentrations as low as 8 μg/mℓ. Betulinic acid was less active than lupeol and ursolic acid against C. elegans. The acetone and dichloromethane extracts were also active against C. elegans with a concentration of 0.31 mg/mℓ resulting in almost 80 % inhibition of larval motility. The use of free-living nematodes may provide information on the activity of potential anthelmintics against parasitic nematodes. Extracts of various medicinal plant species may provide solutions to ill-health of small ruminants caused by parasitic nematodes in poor communities of southern Africa.

scholarly journals A Zinc Metalloprotease nas-33 Is Required for Molting and Survival in Parasitic Nematode Haemonchus contortus

2021 ◽  
Vol 9 ◽  
Author(s):  
Yan Huang ◽  
Jie Wu ◽  
Xueqiu Chen ◽  
Danni Tong ◽  
Jingru Zhou ◽  
...  
Keyword(s):  
Haemonchus Contortus ◽  
Parasitic Nematode ◽  
Large Family ◽  
Parasitic Nematodes ◽  
Larval Stages ◽  
Zinc Metalloprotease ◽  
Molting Process ◽  
Survival And Development ◽  
Free Living Nematode

Molting is of great importance for the survival and development of nematodes. Nematode astacins (NAS), a large family of zinc metalloproteases, have been proposed as novel anthelmintic targets due to their multiple roles in biological processes of parasitic nematodes. In this study, we report a well conserved nas-33 gene in nematodes of clade V and elucidate how this gene is involved in the molting process of the free-living nematode Caenorhabditis elegans and the parasitic nematode Haemonchus contortus. A predominant transcription of nas-33 is detected in the larval stages of these worms, particularly in the molting process. Knockdown of this gene results in marked molecular changes of genes involved in cuticle synthesis and ecdysis, compromised shedding of the old cuticle, and reduced worm viability in H. contortus. The crucial role of nas-33 in molting is closely associated with a G protein beta subunit (GPB-1). Suppression of both nas-33 and gpb-1 blocks shedding of the old cuticle, compromises the connection between the cuticle and hypodermis, and leads to an increased number of sick and dead worms, indicating essentiality of this module in nematode development and survival. These findings reveal the functional role of nas-33 in nematode molting process and identify astacins as novel anthelmintic targets for parasitic nematodes of socioeconomic significance.

scholarly journals Identifications, Characteristics, and Expression Patterns of Small Heat Shock Protein Genes in a Major Mulberry Pest, Glyphodes pyloalis (Lepidoptera: Pyralidae)

2020 ◽  
Vol 20 (3) ◽  
Author(s):  
Jie Chu ◽  
De-lei Jiang ◽  
Meng-wen Yan ◽  
Yi- jiangcheng Li ◽  
Jun Wang ◽  
...  
Keyword(s):  
Heat Shock ◽  
Integrated Management ◽  
Expression Patterns ◽  
Small Heat Shock Protein ◽  
Peak Level ◽  
Functional Differentiation ◽  
Open Reading Frames ◽  
Amino Acid Residues ◽  
Larval Stages ◽  
Glyphodes Pyloalis

Abstract Six candidate sHSP genes were identified from the Glyphodes pyloalis transcriptome. All sHSP genes included full-length open reading frames and shared high similarity with the sequences of other lepidopteran species. These sHSP genes encoded 175–191 amino acid residues, and the predicted proteins had a molecular weight from 19.5 to 21.8 kDa. All GpsHSPs were expressed at lower levels at larval stages. All GpsHSPs were expressed at higher levels at diapaused, prepupal, or pupal stages, suggesting that sHSPs may be involved in metamorphosis in G. pyloalis. In addition to the developmental stage, extreme temperatures can induce variations in the expression of sHSPs genes. All GpsHSPs were significantly upregulated in larvae following exposure to heat shock, except GpHSP21.4 which downregulated at 4 h following exposure to the cold shock treatment. Furthermore, Starvation influenced the expression patterns of GpsHSPs as a function of the duration of food deprivation. Four GpsHSPs increased their expression with time of starvation until reaching to the peak level at 6 d of starvation. Finally, parasitism by the endoparasitoid Aulacocentrum confusum He et van Achterberg (Hymenoptera: Braconidae)-induced fluctuations in the expression of all GpsHSPs, and the expression varied with time after parasitization. Our results from this study strongly suggest functional differentiation within the sHSPs subfamily in G. pyloalis. The present study would provide further insight into the roles of sHSPs in G. pyloalis and novel avenues for promoting integrated management of this pest.

scholarly journals A transcription factor DAF-5 functions in Haemonchus contortus development

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Wenda Di ◽  
Fangfang Li ◽  
Li He ◽  
Chunqun Wang ◽  
Caixian Zhou ◽  
...  
Keyword(s):  
Haemonchus Contortus ◽  
Developmental Stages ◽  
Reproductive Organs ◽  
Bimolecular Fluorescence Complementation ◽  
Parasitic Nematodes ◽  
Small Interfering Rnas ◽  
C Elegans ◽  
Dauer Formation ◽  
And Function

Abstract Background Abnormal dauer formation gene (daf-5), located downstream of the DAF-7 signalling pathway, mainly functions in dauer formation and reproductive processes in the free-living nematode Caenorhabditis elegans. Although the structure and function of daf-5 have been clarified in C. elegans, they still remain totally unknown in Haemonchus contortus, a socio-economically important parasitic nematode of gastric ruminants. Methods A homologue of daf-5, Hc-daf-5, and its inferred product (Hc-DAF-5) in H. contortus were identified and characterized in this study. Then the transcriptional profiles of Hc-daf-5 and the anatomical expression of Hc-DAF-5 in H. contortus were studied using an integrated molecular approach. RNA interference (RNAi) was performed to explore its function in transition from the exsheathed third-stage larvae (xL3s) to the fourth-stage larvae (L4s) in vitro. Finally, the interaction between Hc-DAF-5 and Hc-DAF-3 (a co-Smad) was detected by bimolecular fluorescence complementation (BiFc) in vitro. Results It was shown that Hc-DAF-5 was a member of the Sno/Ski superfamily. Hc-daf-5 was transcribed in all developmental stages of H. contortus, with significant upregulation in L3s. Native Hc-DAF-5 was localized in the reproductive organs, cuticle, and intestine via immunohistochemistry. RNAi revealed that specific small interfering RNAs (siRNAs) could retard xL3 development. In addition, the interaction between Hc-DAF-5 and Hc-DAF-3 indicated that the SDS box of Hc-DAF-5 was dispensable for the binding of Hc-DAF-5 to Hc-DAF-3, and the MH2 domain was the binding region between Hc-DAF-3 and Hc-DAF-5. Conclusions In summary, these findings show that Hc-daf-5 functions in the developmental processes of H. contortus, and this study is the first attempt to characterize the daf-5 gene in parasitic nematodes. Graphical abstract

scholarly journals Cell Death and Transcriptional Responses Induced in Larvae of the Nematode Haemonchus contortus by Toxins/Toxicants with Broad Phylogenetic Efficacy

2021 ◽  
Vol 14 (7) ◽  
pp. 598
Author(s):  
Douglas P. Jasmer ◽  
Bruce A. Rosa ◽  
Makedonka Mitreva
Keyword(s):  
Cell Death ◽  
Small Molecule ◽  
Haemonchus Contortus ◽  
Small Molecule Inhibitors ◽  
Animal Health ◽  
Transcriptional Response ◽  
Parasitic Nematodes ◽  
Transcriptional Responses ◽  
Larval Stages ◽  
Organ Systems

Establishing methods to investigate treatments that induce cell death in parasitic nematodes will promote experimental approaches to elucidate mechanisms and to identify prospective anthelmintics capable of inducing this outcome. Here, we extended recent progress on a method to monitor cell death and to identify small molecule inhibitors in Ascaris suum to Haemonchus contortus, a phylogenetically distant parasitic nematode of significance for both human and agricultural animal health. We utilized a diverse group of small molecule inhibitors referred to as nematode intestinal toxins/toxicants (NITs) coupled with motility, cytological and cell death assays to resolve gross effects on motility and individual cells and organ systems of two H. contortus larval stages in culture. Early transcriptional response evaluation identified NIT-responsive genes and pathways. The scope of death among cells in larvae varied among NITs but shared patterns with A. suum, despite the approach having some limitations due to characteristics of H. contortus larvae. Gene response patterns varied among NITs tested and provided information on the cell targets and pathways affected. Experimental NIT assays provide tools capable of inducing cell death in larval stages of parasitic nematodes, and can resolve many individual cells and organ systems in which cell death can be induced.

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