ABSTRACT
As part of an effort to validate the use of heat shock proteins (HSPs) as biomarkers of exposure to and effects of contaminants, we evaluated the effect of two handling regimens on the induction of HSP 60 and 70 in rainbow trout (Oncorhynchus mykiss). Fish were acclimated to laboratory conditions for several weeks before the beginning of the experiment. Fish were then captured by net, placed in a cooler for 1 h while being transported in a truck, returned to their original tanks, then sacrificed 6 to 8 h later. Tricaine methane sulfonate (MS-222) was used during different phases of handling to reduce handling stress. Heat-stressed fish were included in the experiment as a positive control. Muscle, liver, gills, and heart were analyzed for HSP 60 and 70 by immunoblotting. We found no effect of any handling regimen on the induction of HSPs. These findings suggest that the capture and transport of fish for environmental monitoring purposes should not interfere with the use of stress proteins as biomarkers.
Subject(s)
Biomarkers/analysis , Chaperonin 60/biosynthesis , HSP70 Heat-Shock Proteins/biosynthesis , Oncorhynchus mykiss/physiology , Animals , Gills/chemistry , Immunoblotting , Liver/chemistry , Muscle, Skeletal/chemistry , Reproducibility of Results , Specimen Handling , Tissue DistributionABSTRACT
Crab larvae (Rhithropanopeus harrisiiere exposed to a range of free cupric ion concentrations, [Cu2+], regulated in sea water by a metal chelate buffer system. We found a biphasic relationship between intracellular copper distribution and [Cu2+] in sea water. At [Cu2+] within the ambient range (10-12.4 to 10-10.6 M), cytosolic copper was associated with both metallothionein (MT) and high molecular weight (HMW) ligands, and was independent of external [Cu2+]. At higher [Cu2+], copper was also associated with very low molecular weight (VLMW) ligands, and accumulated in this ligand pool and the MT pool as external [Cu2+] increased. In marked contrast, copper in the HMW ligand pool did not correlate with [Cu2+] in sea water over the entire range of exposures. Reductions in larval growth occurred at greater than estimated ambient [Cu2+] and correlated with copper accumulation in the MT and VLMW pools.
