Comparative gene analysis of Biomphalaria glabrata hemocytes pre- and post-exposure to miracidia of Schistosoma mansoni

The internal defense mechanism of the snail Biomphalaria glabrata during a schistosome infection is activated and mediated via the immune effector cells known as hemocytes. Since resistance and susceptibility to schistosome infection is known to be genetically determined, our interest was to use the...

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Veröffentlicht in:Molecular and biochemical parasitology 2003-02, Vol.126 (2), p.181-191
Hauptverfasser: Raghavan, Nithya, Miller, Andre N, Gardner, Malcolm, FitzGerald, Peter C, Kerlavage, Anthony R, Johnston, David A, Lewis, Fred A, Knight, Matty
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container_end_page 191
container_issue 2
container_start_page 181
container_title Molecular and biochemical parasitology
container_volume 126
creator Raghavan, Nithya
Miller, Andre N
Gardner, Malcolm
FitzGerald, Peter C
Kerlavage, Anthony R
Johnston, David A
Lewis, Fred A
Knight, Matty
description The internal defense mechanism of the snail Biomphalaria glabrata during a schistosome infection is activated and mediated via the immune effector cells known as hemocytes. Since resistance and susceptibility to schistosome infection is known to be genetically determined, our interest was to use the EST approach as a gene discovery tool to examine transcription profiles in hemocytes of resistant snails pre- and post-exposure to Schistosoma mansoni. Comparative analysis of the transcripts suggested that parasite exposure caused an active metabolic response in the hemocytes. The most abundant transcripts were those showing 23–74% similarity to known reverse transcriptases (RT). Further characterization by RT-PCR indicated the RT transcripts were expressed in normal snails, parasite exposed snails, and the embryonic cell line Bge. To determine whether the occurrence of RT transcripts correlates to the presence of functional enzyme activity in the snails, RT assays were performed from both resistant and susceptible snails, pre- and post-exposure to miracidia, using protein extracts from the head–foot and posterior region tissues. Results indicated that in the resistant snail, RT activity was greater in the posterior region than in the head–foot. After exposure, however, RT activity increased dramatically in the head–foot, with peak activity at 24 h post-exposure. The detection of RT activity in B. glabrata was unexpected and the role of this enzyme in the hemocyte-mediated killing of parasites is not yet known. However, identification of this and other transcripts from these cells by the EST approach provides a useful resource towards elucidating the molecular basis of resistance/susceptibility in this snail–host parasite relationship.
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Since resistance and susceptibility to schistosome infection is known to be genetically determined, our interest was to use the EST approach as a gene discovery tool to examine transcription profiles in hemocytes of resistant snails pre- and post-exposure to Schistosoma mansoni. Comparative analysis of the transcripts suggested that parasite exposure caused an active metabolic response in the hemocytes. The most abundant transcripts were those showing 23–74% similarity to known reverse transcriptases (RT). Further characterization by RT-PCR indicated the RT transcripts were expressed in normal snails, parasite exposed snails, and the embryonic cell line Bge. To determine whether the occurrence of RT transcripts correlates to the presence of functional enzyme activity in the snails, RT assays were performed from both resistant and susceptible snails, pre- and post-exposure to miracidia, using protein extracts from the head–foot and posterior region tissues. 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ispartof Molecular and biochemical parasitology, 2003-02, Vol.126 (2), p.181-191
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subjects Amino Acid Sequence
Animals
Bge
Biomphalaria - enzymology
Biomphalaria - genetics
Biomphalaria - parasitology
Biomphalaria glabrata
Blotting, Southern
Cell Line
Ciona intestinalis - enzymology
Ciona intestinalis - genetics
EST
Expressed Sequence Tags
Hemocytes
Hemocytes - parasitology
Hemocytes - physiology
Molecular Sequence Data
Reverse transcriptase
Reverse Transcriptase Polymerase Chain Reaction
RNA-Directed DNA Polymerase - chemistry
RNA-Directed DNA Polymerase - genetics
Schistosoma mansoni - pathogenicity
Sequence Alignment
Sequence Homology, Amino Acid
Transcription, Genetic
title Comparative gene analysis of Biomphalaria glabrata hemocytes pre- and post-exposure to miracidia of Schistosoma mansoni
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