Comparative real-time PCR and enzyme analysis of selected gender-associated molecules inSchistosoma japonicum

Abstract

SUMMARYSchistosomes are complex parasitic helminths with discrete life-cycle stages, adapted for survival in their mammalian and snail hosts and the external aquatic environment. Recently, we described the fabrication and use of a microarray to investigate gender-specific transcription in Schistosoma japonicum. To address transcriptional differences, 8 gender-associated gene transcripts identified previously by the microarray analysis were selected for further study. First, differential transcription patterns were investigated in 4 developmental stages using real-time PCR. Subsequently, we undertook functional analysis of a subset of 4 transcripts encoding metabolic enzymes, so as to correlate gender-associated transcript levels with enzyme activity in protein extracts from adult worms. The 8 characterized molecules serve as a basis for further investigation of differential gene expression during the schistosome life-cycle and for studying the sexual dimorphism of adult worms. Continual refinement and annotation of the microarray used in the current study should support future work on these aspects.

Novel expression profiles of microRNAs suggest that specific miRNAs regulate gene expression for the sexual maturation of female Schistosoma japonicum after pairing

“…Other previous studies have compared differential gene expression between females and males, and similar results have also been reported. For example, egg antigen [ 5 , 40 , 41 ], ferritin-1 [ 5 , 40 , 42 , 43 ], ribosomal proteins [ 5 , 40 , 44 ], ATPase [ 44 , 45 ], cathepsin [ 45 ], extracellular superoxide dismutase [ 5 , 43 , 44 ], cytochrome C oxidase [ 5 ], tyrosinase [ 43 ], mucin-like protein [ 46 ], fs800 [ 47 ], and adenylosuccinate lyase [ 5 ], among others, are often detected in females. These up-regulated genes in paired females are considered to play roles in female sexual maturation and egg production.…”

Developmental gene expression profiles of the human pathogen Schistosoma japonicum

“…In particular, gene expression profiles were more similar within the 4, 6, 6.5 and 7 week old adult parasites, although there was an apparent change in expression from week four (pre-egg laying in female worms) to week six onwards (after the commencement of egg release). This similarity in gene expression likely reflects parasitism of the mammalian blood vessels by the schistosome and the discrete differentiation and development of the males and females [ 4 , 7 , 16 , 17 ]. The eggs, sporocysts and the cercariae also shared similarities in gene expression profiles.…”

Lysosome-associated membrane glycoprotein (LAMP) – preliminary study on a hidden antigen target for vaccination against schistosomiasis

“…All cDNA samples were diluted to 5 ng/μl. Real-time PCR was performed and analysed using previously described protocols 61 using DNA segregation ATPase 62 (Accession No. Smp176580) as the normalising housekeeping gene.…”