A DAF-3 co-Smad molecule functions in Haemonchus contortus development

Di, Wenda; Lü, Ling; Zhang, Ting; Li, Fangfang; He, Li; Wang, Chunqun; Ahmad, Awais Ali; Hassan, Mubashar; Fang, Rui; Hu, Min

doi:10.1186/s13071-019-3855-3

Cited by 14 publications

(24 citation statements)

References 36 publications

(42 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…qRT-PCR analysis confirmed the successful knockdown of the MMP-12 gene in H. contortus L3 in vitro. Previously, successful silencing of the DAF-3 gene by siRNA in H. contortus was reported which is in line with findings of the present study [28]. To evaluate the effects of Hc-MMP-12 siRNA-2 in vivo, a further experiment was performed whereby goats were infected with soaked siRNA-2-treated L3, snRNA-treated L3 and PBS-treated L3.…”

Section: Discussionsupporting

confidence: 91%

“…A gene knockdown technique has been practiced efficiently in parasites by the design and delivery of siRNA of target molecules. In previous studies, RNAi has been used to evaluate the biological functions of different genes in different helminths, for example paramyosin in Trichinella spiralis [23], type V collagen and calcium-regulated heat-stable protein in Schistosoma japonicum [24,25], enolase in Clonorchis sinensis [26], and H11 aminopeptidase and DAF-3 in H. contortus [27,28]. MMPS play important roles in developmental stages [20,21] and the present study was designed to evaluate the biological roles of MMP-12 during different developmental stages of H. contortus in goat.…”

mentioning

confidence: 99%

See 1 more Smart Citation

Haemonchus contortus: siRNA mediated knockdown of matrix metalloproteinase 12A (MMP-12) results in reduction of infectivity

Naqvi

Gao

et al. 2020

Parasites Vectors

View full text Add to dashboard Cite

Background: RNA interference (RNAi) is an important tool to determine the role of genes. RNAi has been widely used to downregulate target molecules, resulting in the reduction of mRNA for protein expression. Matrix metalloprotease 12A (MMP-12) is known to have important roles during embryonic development, organ morphogenesis and pathological processes in animals. However, MMP-12 from Haemonchus contortus has not been characterized.Methods: Haemonchus contortus MMP-12 gene was cloned and recombinant protein of MMP-12 (rHc-MMP-12) was expressed. Binding activities of rHc-MMP-12 to goat peripheral blood mononuclear cells (PBMCs) were assessed by immunofluorescence assay (IFA) and the immuno-regulatory effects of rHc-MMP-12 on cell proliferation and nitric oxide production were observed by co-incubation of rHc-MMP-12 with goat PBMCs. Furthermore, a soaking method was used to knockdown the expression of Hc-MMP12 gene using three siRNA, targeting different regions of the gene and infectivity of effective siRNA on the development of H. contortus was evaluated in goat.Results: rHc-MMP-12 was successfully expressed in an expression vector as well as the tissues of the cuticle of adult H. contortus worms and a successful binding with PBMCs surface were observed. Increased cellular proliferation and nitric oxide production by goat PBMCs was observed in a dose-dependent manner. Quantitative real time PCR (qRT-PCR) results confirmed the successful silencing of Hc-MMP-12 gene in siRNA of 1, 2 and 3 treated third-stage larvae (L3) of H. contortus in vitro. The most efficient qRT-PCR-identified siRNA template was siRNA-2, with a 69% suppression rate compared to the control groups. Moreover, in an in vivo study, silencing of the Hc-MMP-12 gene by siRNA-2 reduced the number of eggs (54.02%), hatchability (16.84%) and worm burden (51.47%) as compared to snRNAtreated control group. In addition, a shorter length of worms in siRNA-2-treated group was observed as compared to control groups. Conclusions:Our results indicate that siRNA-mediated silencing of Hc-MMP-12 gene in H. contortus significantly reduce the egg counts, larval hatchability, and adult worm counts and sizes. The findings of the present study demonstrate important roles of Hc-MMP-12 in the development of H. contortus.

show abstract

Section: Discussionsupporting

confidence: 91%

mentioning

confidence: 99%

Haemonchus contortus: siRNA mediated knockdown of matrix metalloproteinase 12A (MMP-12) results in reduction of infectivity

Naqvi

Gao

et al. 2020

Parasites Vectors

View full text Add to dashboard Cite

show abstract

“…These published findings stimulated our interest in assessing this inhibitor on H. contortus in vitro. The results revealed that SIS3 inhibited the development of L3 larvae in a dosedependent manner, suggesting a role for the Smad3 homologue in larval development in H. contortus, like other members of the TGF-β signalling pathway studied previously in this parasite [12][13][14]. Sequence analysis showed that Hco-DAF-8 has a higher similarity with Hs-Smad3 than the other Smads in H. sapiens at the amino acid level, suggesting that SIS3 specifically inhibits the function of Hco-DAF-8 in H. contortus.…”

Section: Discussionmentioning

confidence: 61%

“…This finding indicates that Cel-daf-8 might play a role in mediating developmental switch (entry into [45,46]. Recent findings from RNAi-based studies also confirm functional roles for genes encoding TGF-β type I receptor (Hc-tgfbr1) [12], TGF-β type II receptor (Hc-tgfbr2) [13] and co-Smad (Hc-daf-3) [14] in the development from the free-living to the parasitic stages of H. contortus.…”

Section: Discussionmentioning

confidence: 62%

“…In the parasite nematode H. contortus, recent studies identified some members of the TGF-β signalling pathway including TGF-β type I receptor [12], TGF-β type II receptor [13] and Co-Smad [14] and evaluated their important roles in larval development from the exsheathed third-stage larvae (L3s) to L4s in intro. As there is no information available on these R-Smads for parasitic nematodes of clade V [15], we took the opportunity of exploring the structure and function of the homologue of Cel-DAF-8 in H. contortus (barber's pole worm), one of the economically most important parasites of small ruminants worldwide [16].…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Identification and characterization of an R-Smad homologue (Hco-DAF-8) from Haemonchus contortus

Gasser

Liu

et al. 2020

Parasites Vectors

Self Cite

View full text Add to dashboard Cite

Background: Smad proteins are essential cellular mediators within the transforming growth factor-β (TGF-β) superfamily. They directly transmit incoming signals from the cell surface receptors to the nucleus. In spite of their functional importance, almost nothing is known about Smad proteins in parasitic nematodes including Haemonchus contortus, an important blood-sucking nematode of small ruminants. Methods: Based on genomic and transcriptome data for H. contortus and using bioinformatics methods, a Smad homologue (called Hco-daf-8) was inferred from H. contortus and the structural characteristics of this gene and its encoded protein Hco-DAF-8 established. Using real-time PCR and immunofluorescence assays, temporal transcriptional and spatial expression profiles of Hco-daf-8 were studied. Gene rescue in Caenorhabditis elegans was then applied to assess the function of Hco-daf-8 and a specific inhibitor of human Smad3 (called SIS3) was employed to evaluate the roles of Hco-DAF-8 in H. contortus development. Results: The features of Hco-DAF-8 (502 amino acids), including conserved R-Smad domains and residues of the L3-loop that determine pathway specificity, are consistent with a TGF-β type I receptor-activated R-Smad. The Hcodaf-8 gene was transcribed in all developmental stages of H. contortus studied, with a higher level of transcription in the fourth-stage larval (L4) females and the highest level in adult males. Hco-DAF-8 was expressed in the platymyarian muscular cells, intestine and reproductive system of adult stages. Gene rescue experiments showed that Hco-daf-8 was able to partially rescue gene function in a daf-8 deficient mutant strain of C. elegans, leading to a resumption of normal development. In H. contortus, SIS3 was shown to affect H. contortus development from the exsheathed thirdstage larvae (L3s) to L4s in vitro. Conclusions: These findings suggest that Hco-DAF-8, encoded by the gene Hco-daf-8, is an important cellular mediator of H. contortus development via the TGF-β signalling pathway. They provide a basis for future explorations of Hco-DAF-8 and associated pathways in H. contortus and other important parasitic nematodes.

show abstract

Toward integrative ‘omics of the barber’s pole worm and related parasitic nematodes

Gasser

Wang

et al. 2020

Infection, Genetics and Evolution

View full text Add to dashboard Cite

A DAF-3 co-Smad molecule functions in Haemonchus contortus development

Cited by 14 publications

References 36 publications

Haemonchus contortus: siRNA mediated knockdown of matrix metalloproteinase 12A (MMP-12) results in reduction of infectivity

Haemonchus contortus: siRNA mediated knockdown of matrix metalloproteinase 12A (MMP-12) results in reduction of infectivity

Identification and characterization of an R-Smad homologue (Hco-DAF-8) from Haemonchus contortus

Toward integrative ‘omics of the barber’s pole worm and related parasitic nematodes

Contact Info

Product

Resources

About