Changes induced by two strains of Vibrio splendidus in haemocyte subpopulations of Mya arenaria, detected by flow cytometry with LysoTracker

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DOIResolve DOI: http://doi.org/10.3354/dao02121
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TypeArticle
Journal titleDiseases of Aquatic Organisms
Volume86
Pages253262; # of pages: 10
SubjectMya arenaria; Vibrio splendidus; Haemocyte subpopulations; Flow cytometry; LysoTracker; Lysosome
AbstractFlow-cytometric characterisation of bivalve haemocytes is usually performed by lightscatter profiles based on size and complexity of the cells. Additional means of characterisation such as specific fluorescent dyes are not commonly used to discriminate cell subpopulations in challenged and unchallenged haemocytes. In the present study, we characterise the changes in haemocyte subpopulations of soft-shell clam Mya arenaria induced by in vivo challenge with 2 strains of Vibrio splendidus by using a fluorescent probe. Responses were measured 24 h after infection with either a local wild strain (7SHRW) or a modification (LGP32-GFP) of a strain associated with oyster mortalities in France (LGP32). Changes in haemocyte subpopulations were analysed using flow cytometry based on 2-parameter scatter profiles and lysosomal content reflected by LysoTracker staining. Forward and side-scatter profiles revealed 2 haemocyte subpopulations: hyalinocytes and granulocytes. Granulocytes exhibited significantly higher levels of lysosomal staining (p < 0.01). Following infection with LGP32-GFP, both subpopulations merged into a single continuous group and their lysosomal content significantly decreased (p < 0.05). Independent modifications after infection were observed in the proportions of subpopulations established by their lysosomal content. While the subpopulation of hyalinocytes had lower levels of lysosomal content after infection, especially with LGP32-GFP (p < 0.001), the subpopulation of granulocytes had similar levels of lysosomes after infection with 7SHRW and significantly decreased levels after infection with LGP32-GFP (p = 0.001). Our data suggest specific modulation of bivalve responses against pathogenic bacteria that would include degranulation.
Publication date
LanguageEnglish
AffiliationNational Research Council Canada; NRC Institute for Nutrisciences and Health
Peer reviewedYes
NPARC number17320717
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Record identifier7e8da9f4-8a48-4c4d-b876-896a8aaaa765
Record created2011-03-17
Record modified2016-05-09
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