REG-5
Mesoscale models of hydrothermal
processes and anthropogenic
admixture transferring in atmosphere
and hydrosphere
Evaluation of biochemical
responses in Palearctic and
Lake Baikal endemic
amphipod species exposed to
toxicity stress
Bolshie Koty, Baikal, Russia
June 25-29, 2007
Marina Protopopova,
Darya Bedulina
Irkutsk State University, Irkutsk
Laboratory of biological systems
adaptation
Lake Baikal:
Flora and fauna 60-80% endemic
The evaluation of
antioxidant enzymes
specific
mechanisms
of stress adaptation
and sHSP
as possible
biomarkers for using
in the toxicological
studiesspecific
of Baikal
biomarkers
endemic species
The aim of research
The objects of research
Freshwater species of amphipods (Crustacea, Amphipoda):
Baikal species
E. cyaneus (Dyb.)
E. verrucosus (Gerstf)
Palearctic species
Gammarus
Gammarus lacustris
lacustris (Sars)
Sars
The biomarkers of stress
Antioxidant enzymes:
• peroxidase (POD)
Toxic stress
• catalase (CAT)
solution of cadmium chloride (CdCl2)
Heat-shock proteins
(HSPs):
• small Heat-shock proteins
(sHSP)
Methods
Measurement of
enzyme activities
POD activity was
measured with guaiacol
as the substrate at 436
nm, pH 5 according to
Bergmeyer (1983)
CAT activity was
measured with hydrogen
peroxide as the substrate
at 240 nm and a pH of 7
according to Aebi (1984).
Spectrophotometric
method
Methods
Measurement of
sHSPs content
SDS- electrophoresis
Western-blotting
Results
POD activities
in the three amphipod
species
exposed to 0,05 mgL-1
CdCl2
Results
CAT activities
in the three amphipod
species
exposed to 0,05 mgL-1
CdCl2
Results
Western blot analysis on
sHSPs of the amphypods
after 24 hours exposition in
different CdCl2
solutions
line 1 – control,
A. E. cyaneus,
B. E. verrucosus, line 2 - 0,05 mgL-1,
line 3 - 0,5 mgL-1,
C. G. lacustris
line 4 – 5 mgL-1.
The chaperon function of sHSPs
stress
The native
protein chain
The denaturated
protein chain
The refolded
protein chain
Conclusion
• CdCl2 causes significant effects of the peroxidase.
• CdCl2 causes induction of the dose-dependent
synthesis of small heat shock proteins.
• All measured parameters varied interspecifically
and depended on the ability of the species to
resist cadmium chloride toxicity.
The synthesis of small heat shock proteins
and peroxidase activity, could be successfully
used as biomarkers for the further studies in
amphipod species including endemics from
Lake Baikal.
The publication:
Evaluation of biochemical responses in Palearctic and
Lake Baikal' endemic amphipod species exposed to
CdCl2 // Ecotoxicology and Environmental Safety, 2007
(in press)
The grants of Russian Foundation
for Basic Research:
№ 06-04-48099-а, 5-04-97239-baikal, 05-04-97263-baikal
Thank you