Ittiprasert Wannaporn, Miller Andre, Myers Jocelyn, Nene Vish, El-Sayed Najib M, Knight Matty
Biomedical Research Institute, Rockville, MD 20852, USA.
Mol Biochem Parasitol. 2010 Jan;169(1):27-39. doi: 10.1016/j.molbiopara.2009.09.009. Epub 2009 Oct 6.
Resistance or susceptibility of the snail host Biomphalaria glabrata to Schistosoma mansoni is determined by the genetics of both the snail and parasite. Although Mendelian genetics governs adult resistance to infection, juvenile resistance and susceptibility are complex traits. In this study, suppression subtractive hybridization was used to construct forward and reverse cDNA libraries to identify genes involved in the immediate response of juvenile resistant (BS-90), non-susceptible (LAC2) snails, and susceptible (NMRI) snails after early exposure to S. mansoni. Expressed Sequence Tags (ESTs) were generated from the repertoire of enriched transcripts. In resistant snails, several ESTs corresponded to transcripts involved in immune regulation/defense response. While no defense related transcripts were found among juvenile susceptible snail ESTs, we detected transcripts involved in negative regulation of biological process/morphogenesis/proliferation. Differential gene expression and temporal regulation of representative transcripts were compared among snails pre- and post-exposure to either normal or attenuated miracidia using quantitative real time RT-PCR. Results showed that several transcripts, such as fibrinolytic C terminal domain, cytidine deaminase, macrophage expressed gene 1, protein kinase C receptor, anti-microbial peptide; theromacin and Fas remained up-regulated regardless of whether or not snails were exposed to normal or attenuated miracidia. While ESTs related to C-type lectin and low-density lipoprotein receptor were induced only by exposure to normal miracidia. By comparing changes in gene expression between resistant and susceptible juvenile snails responding either to normal or attenuated parasites, we can conclude that the transcription of genes associated with the intra-dermal penetration process of the snail host by invading miracidia may need to be taken into account when assessing differential gene expression between resistant and susceptible strains of B.glabrata in relation to S. mansoni exposure.
蜗牛宿主光滑双脐螺对曼氏血吸虫的抗性或易感性由蜗牛和寄生虫的基因共同决定。虽然孟德尔遗传学控制着成虫对感染的抗性,但幼虫的抗性和易感性是复杂的性状。在本研究中,利用抑制性消减杂交技术构建正向和反向cDNA文库,以鉴定早期暴露于曼氏血吸虫后,抗性幼虫(BS-90)、非易感(LAC2)蜗牛和易感(NMRI)蜗牛的即时反应中涉及的基因。从富集转录本库中生成表达序列标签(EST)。在抗性蜗牛中,几个EST对应于参与免疫调节/防御反应的转录本。虽然在易感幼虫蜗牛的EST中未发现与防御相关的转录本,但我们检测到了参与生物过程/形态发生/增殖负调控的转录本。使用定量实时RT-PCR比较了暴露于正常或减毒毛蚴前后蜗牛中代表性转录本的差异基因表达和时间调控。结果表明,无论蜗牛是否暴露于正常或减毒毛蚴,几种转录本,如纤溶酶原C末端结构域、胞苷脱氨酶、巨噬细胞表达基因1、蛋白激酶C受体、抗菌肽;theromacin和Fas仍上调。而与C型凝集素和低密度脂蛋白受体相关的EST仅在暴露于正常毛蚴时被诱导。通过比较抗性和易感幼虫蜗牛对正常或减毒寄生虫反应时基因表达的变化,我们可以得出结论,在评估光滑双脐螺抗性和易感品系与曼氏血吸虫暴露相关的差异基因表达时,可能需要考虑与侵袭性毛蚴在蜗牛宿主体内真皮穿透过程相关的基因转录。