Recovery of primary sporocysts in vivo in the Schistosoma mansoni/Biomphalaria glabrata model using a simple fixation method suitable for extraction of genomic DNA and RNA

Exp Parasitol. 2011 Sep;129(1):11-6. doi: 10.1016/j.exppara.2011.06.003. Epub 2011 Jun 22.

Abstract

Detailed studies of host/parasite interactions are currently limited because in situ gene sequencing or monitoring of parasite gene expression is so far limited to genes presenting a high loci copy number in the Schistosome genome or a high level of expression. Indeed, how to investigate the host parasite molecular interplay when parasites are not directly accessible in vivo? Here we describe a method to circumvent this problem and to analyze DNA and RNA of Schistosoma mansoni during the interaction with its intermediate snail host Biomphalaria glabrata. We propose a technique for improved DNA and RNA extraction from the intra-molluscan stage of the parasite recovered after fixation of infected snails in Raillet-Henry solution. The extractions can be used for genetic analysis, transcription studies and microsatellite genotyping.

MeSH terms

  • Animals
  • Biomphalaria / parasitology*
  • Cricetinae
  • DNA Primers
  • DNA, Helminth / isolation & purification*
  • Disease Vectors
  • Liver / parasitology
  • Mesocricetus
  • Microsatellite Repeats
  • Oocysts / physiology
  • RNA, Helminth / isolation & purification*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Schistosoma mansoni / genetics*
  • Schistosoma mansoni / isolation & purification*
  • Schistosoma mansoni / physiology
  • Tissue Fixation / methods*

Substances

  • DNA Primers
  • DNA, Helminth
  • RNA, Helminth