The effect of a mutagen (N-methyl-N-nitro-N-nitrosoguanidine) on cultured cells from adult Schistosoma japonicum

Ming, Zhenping; Dong, Hui-Fen; Zhong, Qian; Grevelding, Christoph G.; Jiang, Ming-Sen

doi:10.1007/s00436-005-0083-x

Cited by 12 publications

(5 citation statements)

References 15 publications

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“…We recently reviewed the prospects and limitations for schistosome post-genomics, drawing attention to the technical advances that need to be made before the newly generated genomic data can be fully utilized (Wilson et al 2007). Whilst there has been progress in the application of RNA interference to silence genes (Skelly et al 2003), both an homologous schistosome cell line and stable transfection into the germ line appear to be distant prospects, although recent reports of cultured schistosome cells (Ming et al 2006) and virus-induced transfection (Kines et al 2006) are encouraging developments. Furthermore, the release of the fourth draft of the genome annotation (http://www.genedb.org/genedb/ smansoni/) in February 2007 brings a definitive gene list much closer, and with it the feasibility of a genome wide microarray for transcript profiling.…”

Section: Discussionmentioning

confidence: 99%

Patterns of gene expression in schistosomes: localization by whole mountin situhybridization

et al. 2007

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rom the identification of genes to the characterization of their functions and interactions. Developmental biologists have long used whole mount in situ hybridization (WISH) to determine gene expression patterns, as a vital tool for formulating and testing hypotheses about function. This paper describes the application of WISH to the study of gene expression in larval and adult schistosomes. Fixed worms were permeablized by proteinase K treatment for hybridization with digoxygenin-labelled RNA probes, with binding being detected by alkaline phosphatase-coupled anti-digoxygenin antibodies, and BM Purple substrate. Discrete staining patterns for the transcripts of the molecules Sm29, cathepsin L, antigen 10.3 and chorion were observed in the tegument cell bodies, gut epithelium, oesophageal gland and vitelline lobules, respectively, of adult worms. Transcripts of the molecules SGTP4, GP18-22 and cathepsin L were localized to tegument cell bodies and embryonic gut, respectively, of lung schistosomula. We also showed that Fast Red TR fluorescent substrate can refine the pattern of localization permitting use of confocal microscopy. We believe that method of WISH will find broad application, in synergy with other emerging post-genomic techniques, such as RNA interference, to studies focused at increasing our molecular understanding of schistosomes

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Section: Discussionmentioning

confidence: 99%

Patterns of gene expression in schistosomes: localization by whole mountin situhybridization

et al. 2007

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“…All trials were carried out in triplicate and the data were expressed as means±standard errors. Differences in orthogonal test were demonstrated with variance analysis and range analysis [13,14].…”

Section: Experiments Design and Data Analysismentioning

confidence: 99%

Notice of Retraction: Effects of Cultivation Conditions on Growth of Rhodopseudomonas palustris Isolated from Aquaculture Wastewater

Fan

Sun

et al. 2011

2011 5th International Conference on Bioinformatics and Biomedical Engineering

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A kind of Rhodopseudomonas palustris, PSB66, was isolated successfully from wastewater of crab pond in Panjin, Liaoning Province, Northeast China, under anaerobic conditions in our study. We conducted a monofactorial experiment and an orthogonal experiment to find the optimal condition for growth of PSB66. The research results showed that, the quantitative relationship between cell mass concentration and OD can be expressed by liner equation (y=11.511x+0.3378, x was the value of OD678 and y was the cell mass concentration). The optimum medium for culture contained 7% of inocula, 7.2 of pH, 0.2% of NaCl, and 0.25% of yeast extract by range analysis of four factors. R values of four factors indicated that effects of different factors on growth of PSB66 were pH (B)> Yeast extract (D)> Percentage of Inocula (A)> NaCl concentration (C). Value of OD reached 0.515 in the optimum medium while PSB66 was cultured after 120 h, and the cell mass concentration of PSB66 was 6.27× 10 8 units·mL -1 .

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“…First approaches towards cell isolation from schistosomes in the past were successfully performed by mincing adult worms under aseptic conditions and in the presence of trypsin/EDTA. This permitted the access to different kind of cell types suitable for cell culture purposes, but intact tissues and inner organs were disrupted due to mechanical forces [67], [68], [85]. Former attempts to isolate internal organs by a simple protease digestion were not successful (Grevelding, personal communication).…”

Section: Discussionmentioning

confidence: 99%

Whole-Organ Isolation Approach as a Basis for Tissue-Specific Analyses in Schistosoma mansoni

Hahnel

Wilson

et al. 2013

PLoS Negl Trop Dis

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BackgroundSchistosomiasis is one of the most important parasitic diseases worldwide, second only to malaria. Schistosomes exhibit an exceptional reproductive biology since the sexual maturation of the female, which includes the differentiation of the reproductive organs, is controlled by pairing. Pathogenicity originates from eggs, which cause severe inflammation in their hosts. Elucidation of processes contributing to female maturation is not only of interest to basic science but also considering novel concepts combating schistosomiasis.Methodology/Principal FindingsTo get direct access to the reproductive organs, we established a novel protocol using a combined detergent/protease-treatment removing the tegument and the musculature of adult Schistosoma mansoni. All steps were monitored by scanning electron microscopy (SEM) and bright-field microscopy (BF). We focused on the gonads of adult schistosomes and demonstrated that isolated and purified testes and ovaries can be used for morphological and structural studies as well as sources for RNA and protein of sufficient amounts for subsequent analyses such as RT-PCR and immunoblotting. To this end, first exemplary evidence was obtained for tissue-specific transcription within the gonads (axonemal dynein intermediate chain gene SmAxDynIC; aquaporin gene SmAQP) as well as for post-transcriptional regulation (SmAQP).Conclusions/SignificanceThe presented method provides a new way of getting access to tissue-specific material of S. mansoni. With regard to many still unanswered questions of schistosome biology, such as elucidating the molecular processes involved in schistosome reproduction, this protocol provides opportunities for, e.g., sub-transcriptomics and sub-proteomics at the organ level. This will promote the characterisation of gene-expression profiles, or more specifically to complete knowledge of signalling pathways contributing to differentiation processes, so discovering involved molecules that may represent potential targets for novel intervention strategies. Furthermore, gonads and other tissues are a basis for cell isolation, opening new perspectives for establishing cell lines, one of the tools desperately needed in the post-genomic era.

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The effect of a mutagen (N-methyl-N-nitro-N-nitrosoguanidine) on cultured cells from adult Schistosoma japonicum

Cited by 12 publications

References 15 publications

Patterns of gene expression in schistosomes: localization by whole mountin situhybridization

Patterns of gene expression in schistosomes: localization by whole mountin situhybridization

Notice of Retraction: Effects of Cultivation Conditions on Growth of Rhodopseudomonas palustris Isolated from Aquaculture Wastewater

Whole-Organ Isolation Approach as a Basis for Tissue-Specific Analyses in Schistosoma mansoni

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