2. ENVIRONMENTAL ChEMISTRy & TEChNOLOGy 2.1. Lectures

Chem. Listy, 102, s265–s1311 (2008) Environmental Chemistry & Technology
M u s S p r e t u s T r a n s c r i p t E x p r e s s i o n
S i g n a t u r e s i n C o n t a m i n a e d A r e a s
We have used the absolute measurement of mRnA
levels from selected key genes (CYPs and GSTs) to biomonitoring
the exposure and biological effects of pollutants on
free-living nonmodel M. spretus. For this purpose the mRnA
molecules of genes coding for different cytochrome P450 and
glutathione transferases were quantified in mice dwelling at
both the non-contaminated point SOL and contaminated area
PS.
As an example, Fig. 4. shows the concomitant up-regulation
of some Cyp transcripts in M. spretus PS population, as
compared to that at SOL in the Doñana Biological Reserve.
The possibilities of transcription quantification to assess the
level of contamination are clearly demonstrated. In addition,
the absolute Cyp transcript expression signature is depending
on the type of contaminant (Fig. 5.), which can use to identify
the nature of contamination under consideration.
Fig. 5. Transcripts expression signature related to the type of
contaminants. y-axis, nRNA molecules/pg total RNA
Fig. 6. Protein expression of liver cytosolic extract from Mus
spretus
P r o t e o m i c s S t u d y o f M u s s p r e t u s
The homology between M. musculus and M. spretus,
at DnA sequence level allows the proteomics evaluation of
s306
protein expression differences caused by contamination episodes
in Mus spretus, but usinf the gnetinc sequence of Mus
musculus in protein identificacion by MALDI-TOF-PMF. A
comparative study at proteomic level of cytosolic fractions
of liver from M. spretus sampled in Doñana and Domingo
Rubio stream were performed analysing the extracts by 2-DE
and searching the protein expression differences. Over 2500
spots were resolved in the pH range 4–7 and 14–70 kDa M r .
Image analysis of the gels yielded 36 spots with significantly
altered expression. Of them, 16 proteins were identified by
MALDI-TOF-PMF and heterologous search against Mus
musculus databases. When this approach is applied to contaminated
and non-contaminated points a clear diference in
spots intensities corresponding to differentially expressed
protreins was observed, which can be used for the environmental
pollution assessment of proteomics.
M e t a l l o m i c s A p p r o x i m a t i o n t o t h e
M o u s e M u s m u s c u l u s
In this study the presence of unknown metallobiomolecules
in M. musculus was studied for the first time. Firstable,
a general evaluation of the presence of total concentration of
metal in different organs of the mouse (lung, liver, spleen,
kidney, brain, testicle, heart and muscle) was performed.The
experiments were carried out on inbred M. musculus specimens.
An you can see the elements considered. Fe is the element
most abundant in the different organs with an averaged
concentration of 7,500 µg dm –3 in thecytosolic extracts. Other
elements are Zn (about 2,300 µg dm –3 ), Cu (550 µg dm –3 ),
ni (165 µg dm –3 ), Se (145 µg dm –3 ), and a toxic element such
as Pb (26 µg dm –3 ). These elements are differentially distributed
in the organs, for example, Cu and Zn are mainly present
in the liver, but Pb in the muscle. An similar comments
can be addressed to the other elements.
Metal-biomolecules profiles was obtained with ICP-MS
as metal tracer, using couplings with size exclusion chroma-
Fig. 7. SEC-ICP-MS metal profiles in Mus musculus corresponding
to Cu and Pb. hMM size exclusion column for 3,000 to
70,000 Da; LMM, size exclusion column for 300 to 10,000 Da