Transcript Slajd 1

2011 Student Practice in JINR Fields of Research
Dubna, Russia
July 13 – 29, 2011
Studying of regularity of
programmed cell death
after influence of demages
factors
University of Wroclaw
Speaker: Karolina Socha
Wroclaw Medical University
Our Group:
Laboratory of Radiation Biology (LRB)
Joint Institute for Nuclear Research
Supervisor: Ivanka Ravnachka
JINR Group:
Elena Baranova
Maria Saveleva
Student Group:
Tatiana Alexandru (Romania)
Karolina Socha (Poland)
Katarzyna Shteklova (Czech Republic)
Outline:
1.
2.
3.
Introduction.
Materials and methods.
Results.
1. Introduction
Studying of regularity of programmed cell death after influence of demages factors
Relation between LET (Linear Energy
Transfer) and the action type
Direct action is predominant
with high LET radiation, e.g.
alpha particles and neutrons.
Indirect action
is predominant with low
LET radiation, e.g. X and
gamma rays.
1. Introduction
Studying of regularity of programmed cell death after influence of demages factors
Types of radiation-induced lesions in DNA
Base damage
Single-strand breaks
Double-strand breaks
1. Introduction
Studying of regularity of programmed cell death after influence of demages factors
Consequences of DSB induction in the cell genome
Studying regularities
in the induction of
apoptosis and DNA
double-strand
breaks as the main
initiating events in
apoptotic cell death
after the action of
radiations with
different physical
properties.
Apoptosis
Neoplastic
transformation
of a cell
DNA
double-strand
break
Chromosome
aberrations
Mutations
1. Introduction
Studying of regularity of programmed cell death after influence of demages factors
Differences between necrosis and apoptosis
Necrosis
Accidental uncontrolled cell death
leads to lysis of cell, inflammatory
responses and serious health
problems.
Apoptosis
Figure 1.
Major diffrences between apoptosis and necrosis.
Programmed cell death
occurs normally during development
and aging and as a homeostatic
mechanism to maintain cell populations
in tissues; defense mechanism (immune
reactions or when sells demaged by
disease or noxious agents (Norbury and
Hickson, 2001).
Studying of regularity of programmed cell death after influence of demages factors
Materials and methods
1.
2.
3.
4.
Figure 2. Fluorescent
microscope (Carl Zeiss).
5.
Seperate lymphocytes from whole
human blood.
Add Hydrogen Peroxide.
Do preparations stained with mix
of dyes (ethidium bromide and
arcidine orange).
Use fluorescent microscope for
tracing morphological changes in
preparations.
Master the morphological method
of detecting various forms of cell
death.
2. Materials and methods
Studying of regularity of programmed cell death after influence of demages factors
Apoptosis visualization
Clasification criteria:
1.
2.
Bright green cells (irregularly shaped nucleus with
heterogeneously poited chromatin) → living cells
Green or red cells (intact chromatine structure) →
necrotic cells
3.
Green or red cells (characteristic nuclear morphology: a
rounded nucleus with collapsed chromatin) → apoptotic
cells
Figure 3.
Different forms of cell death
Studying of regularity of programmed cell death after influence of demages factors
Results
necrotic
apoptosis
living
H2O2
Apoptotic cell induction after
adding H2O2
30
0h
24h
48h
80
60
%
60
0
40
Tim
e(
70
20
h)
0
50
necrotic
apoptosis
living
Control
40
30
20
60
10
30
0
control
H2O2
0
48
24
Tim
e(
h)
0
%
Apoptotic cells, %
60
3. Results
Studying of regularity of programmed cell death after influence of demages factors
Apoptotic cell induction after 
iradiation
 -rays
Apoptotic cells [%]
80
60
40
20
0
0
1
2
3
4
5
6
7
8
9
10
Dose [Gy]
0h
24 h
48 h
Studying of regularity of programmed cell death after influence of demages factors
Thank you for attention!