In situ hybridisation for flow cytometry: a molecular method for monitoring stress-gene expression in hemolymph cells of oysters.
In the present study, we developed a molecular method for flow cytometry to detect the effects of environmental factors on the stress-response in immunocompetent cells of the European flat oyster, Ostrea edulis. Stress-generating conditions were applied to individuals acclimated in the laboratory to environmental salinity and temperature. Oligonucleotidic probes were applied to quantify the expression of HSP/C70 or metallothionein genes. After a heat-stress, a response was reflected in hemocytes by an increased amount of HSP/C70 mRNA and quantitative changes in HSP/C70 protein expression. The technique of in situ hybridisation described here also allowed to quantify the expression of metallothionein mRNA in oysters exposed to a heavy metal-exposure. The detection of stress protein markers by using such quantitative methods could be applied in contamination studies in oysters and other bivalves where monitoring the status of hemolymph cells is required.
Corporeau
Charlotte
C
Laboratoire des Sciences de l'Environnement Marin (LEMAR) UMR 6539, Institut Universitaire Européen de la Mer, Technopôle Brest-Iroise, F-29280 Plouzane, France. charlotte.corporeau@univ-brest.fr
Auffret
Michel
M
D000818:Animals; D004784:Environmental Monitoring / Q000379:methods; D005434:Flow Cytometry / Q000379:methods; D020869:Gene Expression Profiling / Q000379:methods*; D018840:HSP70 Heat-Shock Proteins / Q000187:drug effects / Q000235:genetics*; D006434:Hemocytes / Q000187:drug effects / Q000378:metabolism*; D006458:Hemolymph / Q000166:cytology / Q000187:drug effects; D006358:Hot Temperature; D017403:In Situ Hybridization / Q000379:methods; D008668:Metallothionein / Q000187:drug effects / Q000235:genetics*; D019216:Metals, Heavy / Q000633:toxicity; D010125:Ostreidae / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis; D013312:Stress, Physiological / Q000235:genetics*; D014873:Water Pollutants / Q000633:toxicity
D018840:HSP70 Heat-Shock Proteins; D019216:Metals, Heavy; D012333:RNA, Messenger; D014873:Water Pollutants; D008668:Metallothionein