Studying of regularity of programmed cell death after influence of demages factors
Speaker: Karolina Socha
2011 Student Practice in JINR Fields of Research Dubna, Russia
July 13 – 29, 2011
University of Wroclaw
Wroclaw Medical University
Our Group:
Laboratory of Radiation Biology (LRB)
JINR Group:Elena BaranovaMaria Saveleva
Student Group:Tatiana Alexandru (Romania)Karolina Socha (Poland)Katarzyna Shteklova (Czech Republic)
Joint Institute for Nuclear Research
Supervisor: Ivanka Ravnachka
1. Introduction.2. Materials and methods.3. Results.
Outline:
Relation between LET (Linear Energy Transfer) and the action typeDirect 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
NeoplasticNeoplastictransformationtransformation
of a cellof a cell
ChromosomeChromosomeaberrationsaberrations
MutationsMutations
ApoptosisApoptosis
DNADNAdouble-stranddouble-strand
breakbreak
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.
Consequences of DSB induction in the cell genome
1. Introduction Studying of regularity of programmed cell death after influence of demages factors
Figure 1. Major diffrences between apoptosis and necrosis.
NecrosisAccidental uncontrolled cell death
leads to lysis of cell, inflammatory
responses and serious health
problems.
Apoptosis
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).
Differences between necrosis and apoptosis1. Introduction Studying of regularity of programmed cell death after influence of demages factors
Materials and methods
Figure 2. Fluorescent microscope (Carl Zeiss).
1. Seperate lymphocytes from whole human blood.
2. Add Hydrogen Peroxide.
3. Do preparations stained with mix of dyes (ethidium bromide and arcidine orange).
4. Use fluorescent microscope for tracing morphological changes in preparations.
5. Master the morphological method of detecting various forms of cell death.
Studying of regularity of programmed cell death after influence of demages factors
Apoptosis visualizationClasification criteria:
1. Bright green cells (irregularly shaped nucleus with heterogeneously poited chromatin) → living cells
2. 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 factors2. Materials and methods
0
20
40
60 0
30
60
necrotic apoptosis living
%
Time (h)
H2O
2
0
24
480
30
60
necrotic apoptosis living
%
Time (h)
Control
Apoptotic cell induction after adding H2O2
Studying of regularity of programmed cell death after influence of demages factors
Results
0 1 2 3 4 5 6 7 8 9 100
20
40
60
80
Apo
ptot
ic c
ells
[%]
Dose [Gy]
-rays
Studying of regularity of programmed cell death after influence of demages factors3. Results
Apoptotic cell induction after iradiation
Thank you for attention!
Studying of regularity of programmed cell death after influence of demages factors