Jean-Paul Bourdineaud*1, Nadge Bellance2, Giovani Bnard2,

Daniel Brthes3, Masatake Fujimura4, Patrice Gonzalez1, Aline Marighetto5,

Rgine Maury-Brachet1, Ccile Mormde5, Vanessa Pdron1, Jean-
Nicolas Philippin5, Rodrigue Rossignol2, William Rostne6, Masumi Sawada4
and Muriel Laclau1,3,5
 BACKGROUND

The problem of mercury contamination increases with

time in Wayana Amerindian population.
In 1997, 64% population presented a hair mercury
concentration exceeding the safety limit and have adverse
effect on brain development and neurological disorder.
In 2005, the proportion reached among 84% population
Methylmercury is a neurotoxic compound, which has
been shown to be the cause of the Minamata disease
Human exposure to methyl mercury has been linked to
fish and shellfish consumption
 Calculate the mercury concentration in mice
tissues
Knowing the differential gene expression after
consuming diet contain contaminated fish in
mice
Knowing the differential muscle respiration rate
after consuming diet contain contaminated fish
fish in mice
Knowing the differential anxiety level after
consuming diet contain contaminated fish in mice
 This rats
Male research was an
experimental study by using mice
Age 3 weeks
c57B1/6 model.
Selection
Weight of mice
: 8.2 0.1based
g on a
closer phylogenetic relationship
with humans so that the research
result could be describe the
impact of mercury on human
body system
Invivo study using four treatmen
group
THE FISH REGIMEN THAT GIVEN TO THE MICE WOULD
AS CLOSELY AS POSSIBLE MIMIC A HUMAN
CONTAMINATION ( WAYANAS CONTAMINATION MODE
REGIMEN DIET
Flesh originate from fish contaminated by mercury in their
natural habit and caught in Sinnamary Rver in French Guiana

Highly contaminated by meHg

Hoplias Aimara species, 27% Wayana dietary mercury intake
10.7% total flesh population
consumption

mice age
Average weight of adult men food25-45
pellets had
: 60 kgto contain 5 ng
Hg/g brought
Average mercury consumption by dry
: 60g/day
Mercurial contaminationfish flesh supplementation.
pressure = 1 ng/g/day The H.
aimara fish whose
flesh was used was caught in French
For mice weight 25 g, Guiana in the Sinnamary River, known to
mercury consumption be contaminated by methylmercury
should be 25 ng mostly originating from the Petit-Saut
Brought by a mean hydroelectric reservoir [ The dry flesh of
consumption of 5 g this animal contained 5 g Hg/g. Thus, a
pellets diet containing 0.1% of this fish flesh
could mimic Wayana's contamination
 MICE TREATMENT
Group 1 Group 2 Group 3 Group 4

FED FOR ONE MONTH

CROZZ MAZE TEST

KILL THE MICE BY

DECAPITATION

CUT THE
COLLECT THE DISSECT THE
GASTROCNEMIUS
BLOOD TISSUE
MUSCLE

MERQURY QUANTIFICATION RESPIRATION RATE

GENE EXPRESSION ANALYSIS
TES CROZZ MAZE
 MERCURY GENE EXPRESSION
QUANTIFICATION ANALYSIS
Analyses were carried out Total RNAs were extracted from
automatically after thermal 40 mg of fresh hippocampus,
decomposition at 750C under an
oxygen flow
liver, kidney, and muscle tissues

First-strand cDNA was synthesized

The detection limit was 0.01 ng Hg from 5 g total RNA

Thevalidity of the analytical methods

was checked duringeach series of Real time PCR
measurements using three standard
biological reference materials
http://www.bmb.leeds.ac.uk/teaching/icu3/lecture/20/index.htm
Impact of fish-
containing diets
on anxiety level

mice fed 1 month with diets containing 0.1% or 1% of fish-flesh developed higher
anxiety state behaviors compared to mice fed with control diet.
mice fed with the 7.5% fish-containing diet did not exhibit any statistically relevant
differences in anxiety-like state behavior compared to controls
MERCURY BIOACUMULATION WITHIN MICE
ORGAN
After 34 days feeding with fish meat contaminated with
mercury, there were no differences in body weight among
the four groups of rats.

2. At the lowest dose of fish (0.1% dose), accumulation of

mercury in the hair 80 times higher than mercury levels in
control. At the highest dose of fish (7.5%), rat hair
containing 10 3g Hg / g similar to those found in 84%
of the population Wayana Amerindians. Hair.

Kidney is the organ most contain mercury, and on dietary

regimens containing fish meat doses 0.1 and 1%
contained mercury concentrations comparable to those
on the hair.

In all organs observed mercury concentration gradient is

proportional to ii meat and fish in accordance with
intesintas mercury contamination.
Impact of fish-
containing diets on
gene expression
1. The highest basal expression of the COX1 gene, Sod2, Bax and Fos found in skeletal muscle. The highest
basal expression of genes found in the kidney Sod3. The highest basal expression of genes in liver Gst4
obtained. The highest basal expression of genes and MT2 Mt1 obtained in the hippocampus.

2. In fish, dietary regimens with the lowest dose (0.1%), ekspreis differential observed for COX1 and MT2
genes in the liver and for Fos in the kidney, but none of the genes in a test on hpokampus berespins and
skeletal muscle.

3. On a diet regimen with a dose of fish menenah (1%), in the liver occurs differential gene expression
levels of COX1, gst4, sod2, sod3 and MT2 that may indicate the occurrence of oxidative stress and
effects on mitochondrial (COX1 increased 16-fold).

4. On a diet regimen with a dose of fish menenah (1%), in the liver occurs differential gene expression
levels of COX1, gst4, sod2, sod3 and MT2 that may indicate the occurrence of oxidative stress and
effects on mitochondrial (COX1 increased 16-fold).

5. 5. Response genes can be observed for concentrations of mercury that accumulates in the liver tissue at
least 0.15ppm, 1.4ppm in the kidney and 0.4 ppm in the hippocampus.
Respiratory rates assayed on skinned muscle fibers
 DISCUSSION
a vegetarian diet containing as little
as 0.1% of mercury-contaminated
fish is able to trigger, after only one
month of exposure, bioenergetical
disorders in skeletal muscles, a
genetic response in liver and
kidneys, and an increase in the
anxiety-driven behavior of mice
demonstrating that the aimara flesh
is harmful
 DISCUSSION
1. methylmercury is the only known toxic compound
contaminating the food web of the Sinnamary River, and
apart from clandestine gold mining activities, no sources
of organic xenobiotics have been recorded so far in this
part of the Amazonian jungle.
2. the mercury accumulation in mice tissues is dependent on
the diet fish content.
3. gene expression studies are now powerful enough to
discriminate and classify toxicants on the basis of unique
gene expression profiles induced by putative toxic actions
4. our results on muscle mitochondrial respiration are fully
concordant with the long-known effects of methylmercury
on mitochondria of human and rat liver
 These comparative patterns of gene
expression data indicate that when shifting
from chronic to acute exposure the
intensity of gene response does not
increase as might have been expected but
rather that the qualitative nature of the
gene response is completely changed
resulting in a modified tissue metabolism
suggest a model in which low doses of
pollutant cause mild effects compatible
with life, allowing animal resistance-
through adaptive response, whereas higher
doses trigger acute effects threatening
animal's life, resulting in general stress
instead of adaptive response
 Mt I

Divalent mercury

Mt II

Induce free radicals

Methyl mercury
intoxication Gene p 53,
Induce the
transcription factor
apoptotic pathway
c Fos, bax/bcl2

Antioxidant
enzymes SOD, GSt
Inactivate the
enzymes
Respiration chain
enzyme
 gene expression studies are now powerful enough to
discriminate and classify toxicants on the basis of unique gene
expression profiles induced by putative toxic actions

Methyl mercury is more specific to Methyl mercury is specific to cause

cause nervous system disruption mitochondrial function disruption

Alteration of Reduce State Inactivation of

Anxiety test
gene 3 respiration cytochrome c
result
expression rate oxidase
CONCLUSION
 THANK YOU
FOR YOUR ATTENTION