Validation of Suitable Reference Genes for Expression Normalization in Echinococcus spp. Larval Stages

Ferreira, Henrique Bunselmeyer; Zaha, Arnaldo

doi:10.1371/journal.pone.0102228

Cited by 25 publications

(18 citation statements)

References 34 publications

(67 reference statements)

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…In this study, FABP genes are downregulated during E. granulosus adult worm development. Antigen B has both upand downregulated genes, in agreement with previous works[33,34].…”

supporting

confidence: 92%

Transcriptomic analysis of the early strobilar development of Echinococcus granulosus

Debarba

Monteiro

Gerber

et al. 2018

Preprint

Self Cite

View full text Add to dashboard Cite

Background: Echinococcus granulosus has a complex life cycle involving two mammalian hosts. The transition from one host to another is accompanied by changes in gene expression, and the transcriptional events that underlie these processes have not yet been fully characterized.Results: In this study, RNA-seq is used to compare the transcription profiles of four time samples of E. granulosus protoscoleces in vitro induced to strobilar development. We identified 818 differentially expressed genes, which were divided into eight expression clusters formed over the entire 24 hours time course and indicated different transcriptional patterns. An enrichment of gene transcripts with molecular functions of signal transduction, enzymes and protein modifications was observed with progression of development. Conclusion:This transcriptomic study provides insight for understanding the complex life cycle of E. granulosus and contributes for searching for the key genes correlating with the strobilar development, providing interesting hints for further studies. 2

show abstract

“…In this study, FABP genes are downregulated during E. granulosus adult worm development. Antigen B has both upand downregulated genes, in agreement with previous works[33,34].…”

supporting

confidence: 92%

Transcriptomic analysis of the early strobilar development of Echinococcus granulosus

Debarba

Monteiro

Gerber

et al. 2018

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…The qPCR primer pairs of Eg-TSP1 and elongation factor 1 alpha (EF1α, an optimized housekeeping gene in Echinococcus spp. [ 35 ]) were given in Additional file 1 . All amplifications were performed as described above, except for the annealing temperatures: 53.2 °C for Eg-TSP1 and 57.8 °C for EF1α.…”

Section: Methodsmentioning

confidence: 99%

Molecular insights into a tetraspanin in the hydatid tapeworm Echinococcus granulosus

Song

Xie

et al. 2015

Parasites Vectors

View full text Add to dashboard Cite

BackgroundCystic echinococcosis (hydatid disease), caused by the tapeworm Echinococcus granulosus (class Cestoda; family Taeniidae), is a neglected tropical disease that results in morbidity and mortality in millions of humans, as well as in huge economic losses in the livestock industry globally. Proteins from the tetraspanin family in parasites have recently become regarded as crucial molecules in interaction with hosts in parasitism and are therefore suitable for the development of vaccines and diagnostic agents. However, no information is available to date on E. granulosus tetraspanin.MethodsIn this study, a uroplakin-I-like tetraspanin (Eg-TSP1) of E. granulosus was cloned and expressed in E. coli. The immunolocalization of Eg-TSP1 in different life stages of E. granulosus was determined using specific polyclonal antibody. The antibody and cytokine profiles of mice that immunized with recombinant Eg-TSP1 (rEg-TSP1) were measured for the immunogenicity analysis of this protein. Additionally, we use RNA interference method to explore the biological function of Eg-TSP1 in larva of E. granulosus.ResultsImmunofluorescence analysis showed that endogenous Eg-TSP1 mainly localized in the tegument of larvae and adults. Significantly elevated levels of antibodies IgG1 and IgG2a and of cytokines IFN-γ and IL-12 were observed in the sera of mice after immunization with rEg-TSP1, suggesting a typical T helper (Th)1-mediated immune response elicited by rEg-TSP1. On further probing the role of Eg-TSP1 in E. granulosus by RNA interference, we found that a thinner tegmental distal cytoplasm was induced in protoscoleces treated with siRNA-132 compared to controls.ConclusionsThis is the first report characterizing a tetraspanin from the tapeworm E. granulosus. Our results suggest that Eg-TSP1 is associated with biogenesis of the tegument and maintenance of structural integrity of E. granulosus and could therefore be a candidate intervention target for control of hydatid disease.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-015-0926-y) contains supplementary material, which is available to authorized users.

show abstract

“…EgrG_000982200), and cyclophilin ( eg-cyp , GeneDB no. EgrG_000920600) were adapted from Espínola et al ( 2014 ). Table 2 summarizes the characteristics of the primers.…”

Section: Methodsmentioning

confidence: 99%

Comparison of ex vivo harvested and in vitro cultured materials from Echinococcus granulosus by measuring expression levels of five genes putatively involved in the development and maturation of adult worms

et al. 2016

View full text Add to dashboard Cite

Parts of the natural life cycle of Echinococcus granulosus can be retraced in vitro such as the development of protoscoleces into semiadult worms with three or more proglottids, or the redifferentiation of in vitro cultured protoscoleces into metacestode-like cystic structures. Most in vitro generated samples share—at the microscopical level—high similarities with those naturally grown, but developmental differences have also been documented, such as missing egg production in in vitro grown adults or unusual bladder/vesicle formation in protoscoleces cultured into the metacestode direction. The aim of the present study was to explore how far different in vitro generated stage-specific materials/structures match the natural situation on the transcriptome level, based on testing five exemplarily chosen different genes: the frizzled receptor eg-fz4 (posterior marker), the FGF receptor-like factor eg-fgfrl (anterior association), the cell differentiation protein eg-rcd1 (part of the CCR4-NOT complex, a key regulator of eukaryotic gene expression), the rapidly accelerated fibrosarcoma serin/threonin kinase eg-braf (part of the MAPK pathway involved, e.g., in EGF signaling) and the co-smad eg-smadD (downstream factor of TGFβ/BMP2/activin signaling). These genes—tested via qPCR—were selected such as to allow a discussion on their potential role in the development of E. granulosus into the adult stage. Thus, testing took place with three ex vivo isolated samples, namely (i) egg-containing adult worms, (ii) invaginated protoscoleces, and (iii) protoscolex-free germinal layer tissue. Respective data were compared (a) with in vitro generated metacestode-like microcysts developed from protoscolices, and (b) different development stages of protoscoleces in vitro cultured toward adult maturation. As a finding, only eg-smadD and partially eg-fz4 showed high expression similarities between ex vivo harvested and in vitro cultured E. granulosus, thus suggesting a putative role in adult maturation. Conclusively, the fact of using “only” five genes did not allow answering the question if ex vivo and in vitro materials are similar on the transcriptome level. Another experimental restriction arises from different growth conditions of the in vitro cultured materials, and comparing these to the ex vivo harvested ones. Future experiments may solve the problems by using fully standardized E. granulosus sample collection and fully standardized culture conditions.

show abstract

Validation of Suitable Reference Genes for Expression Normalization in Echinococcus spp. Larval Stages

Cited by 25 publications

References 34 publications

Transcriptomic analysis of the early strobilar development of Echinococcus granulosus

Transcriptomic analysis of the early strobilar development of Echinococcus granulosus

Molecular insights into a tetraspanin in the hydatid tapeworm Echinococcus granulosus

Comparison of ex vivo harvested and in vitro cultured materials from Echinococcus granulosus by measuring expression levels of five genes putatively involved in the development and maturation of adult worms

Contact Info

Product

Resources

About