Chromosome Aberrations

CHROMOSOME
ABERRATIONS
AND
HUMAN LYMPHOCYTES
A LITTLE BACKGROUND
Traditionally the study of radiation damage
on chromosomes was principally conducted
on plant cells (e.g. Tradescantia paludosa)
 Study on mammalian cells hampered by large
number of chromosomes per cell and small size
of nucleus
 Plants contain fewer and generally much larger
chromosomes
Study of chromosome aberrations through the
effects of ionizing radiation described in terms
of their appearance at first metaphase after
exposure to radiation
CELL CYCLE
From: Brum GD, et al (1989). Biology:Exploring Life pg. 156

HOW IT HAPPENS
When cells are irradiated, breaks are
produced in the chromosomes
Broken ends appear to be ‘sticky’ and can
rejoin with any other sticky end
Once breaks are produced, different
fragments may behave in a variety of ways
 Breaks may rejoin in their original configuration
 Breaks may fail to rejoin and give rise to a
deletion
 Broken ends may resort and rejoin other broken
ends and give rise to chromosomes that appear
to be grossly distorted
TYPES OF ABERRATIONS
ABERRATIONS SEEN AT METAPHASE ARE OF 2 CLASSES
 Chromosome Aberrations
- result if cell is irradiated early in interphase (G1),
before chromosome material has been duplicated
- during DNA synthetic phase that follows, this
strand of chromatin lays down an identical strand
next to itself
Chromatid Aberration
- result if cell is irradiated later in interphase after
DNA has doubled (G2) and chromosome consist of
2 strands of chromatin
- break that occurs in a single chromatid arm, leaves
opposite arm of same chromosome undamaged
EXAMPLES OF ABBERATIONS
Many types of aberrations and
rearrangements are possible
3 Types are lethal to the cell
 Dicentric, Centric Ring (chromosome
aberrations) and Anaphase Bridge (chromatid
aberration)
2 Important Non-Lethal Rearrangements
 Symmetric translocations and small deletions
(interstitial and terminal)
 Associated with several human malignancies via
activation of oncogenes or loss of suppressor
genes
LETHAL ABERRATIONS
All 3 represent gross chromosomal distortions
Dicentric (or Tricentric)

 Involves interchange between 2 separate
chromosomes
 If break occurs in each one early in interphase and
sticky ends are close together they may join
 this interchange is replicated during DNA synthesis
and results in a distorted chromosome with 2 (or 3)
centromeres and an acentric fragment
DICENTRIC CHROMOSOME
From: Radiobiology for the Radiologist, pg 24

Centric Ring
 Break induced by radiation in each arm of a
single chromatid early in cell cycle
 The sticky ends may rejoin to form a ring with a

centromere and an acentric fragment
 Later during DNA synthetic phase the

chromosome is replicated
CENTRIC RING

Anaphase Bridge
Results from breaks that occur late in cell cycle (G2),
after chromosome has replicated
Breaks may occur in both chromatids of the same
chromosome, sticky ends may rejoin incorrectly to
form a sister union
At anaphase, when the 2 sets of chromosomes move
to opposite poles, the section of chromatin between
the centromeres is stretched across between the
poles, hindering separation into new daughter cells
Difficult to see in human cell cultures since bridge is
only evident at anaphase
ANAPHASE BRIDGE

Symmetric Translocation
 Involves break in 2 pre-replication chromosomes,
with broken ends being exchanged between the 2
chromosomes
 Divided into pericentric (includes centromere) and
paracentric (confined to 1 chromosome arm)
inversions
 Difficult to see in conventional preparation, but easy
to observe with fluorescent in situ hybridization
(FISH)
 A translocation is associated with several human
malignancies by the activation of an oncogene (e.g.
Burkitt’s lymphoma)
Small Deletion
 Terminal deletion:
 Loss of genetic material from end of chromatid
 Not possible to distinguish between non-sister
union isochromatid and chromosome-type
 Interstitial deletion
 Minute: very small deletion; show as small paired
dots
 Acentric ring: larger interstitial deletion; show as
acentric rings
 Deletion may be associated with
carcinogenesis if loss of material includes a
suppressor gene
From: Biological Dosimetry, pg 18
HUMAN LYMPHOCYTES AS A
BIOMARKER
Chromosomal aberrations in peripheral lymphocytes
currently most fully developed biological indicator of
exposure to ionizing radiation
In vitro and in vivo irradiation of blood lymphocytes
produces similar yields of chromosome damage per
rad, so that the observed of aberrations in exposed
persons can be related to the dose by comparison
with an in vitro produced dose-response curve
In circulation they are generally non-dividing and
accumulate damage typical of that caused by
irradiation in the G0/G1 cell cycle stage
RADIATION EFFECTS ON
LYMPHOCYTES
Undamaged cells contain 46 chromosomes,
each with 1 centromere
Damaged cells may display a number of
types of aberrations including dicentrics,
centric rings, acentric fragments and
translocations, all of which may be related to
radiation dose
From: Biological Dosimetry, pg 16
DICENTRICS AS A BIOMARKER
For biological dosimetry the dicentric historically has
been the aberration of choice
 most reliably scored, easily seen in
chromosome spread
 low level of occurrence in unirradiated
persons (about 1/1000 cells)

 10x more common than rings and about as
common as excess fragments

 Less reliably scored
 Have a higher occurrence (1/300 cell) in
unirradiated persons
SCORING LYMPHOCYTE
ABERRATIONS
Lymphocytes may be stimulated to divide in
vitro by adding phytohemagglutinin (PHA)
Stopped at their 1st metaphase by addition of
Colcemid after about 45 hrs of culture
Slides containing metaphase spreads stained
with Giesma or FISH probes and scored
Contamination with 2nd division cells made by
incorporating bromodeoxyuridine (BrdU) into
the culture
In suspected exposure case, typically 500
cells are scored
DOSE-RESPONSE CURVES
Y=c+D+D2
where Y is yield of dicentrics/cell, D is dose(Gy) and c,, are fitted constants
From: The use of Chromosomal Aberrations in Human Lymphocytes for Biological Dosimetry, pg S41
LIMITATIONS OF SCORING
DICENTRICS
Major limitation of using dicentrics for
dosimetry is loss of lymphocytes from the
blood
 Yield of measured dicentrics after an irradiation
would decrease with time (1/2 life is about 3 yrs),
although this is uncertain
When replaced by stem cell division, new
lymphocytes will tend not to contain dicentrics
due to elimination at anaphase
 unstable aberration since it is lethal to the cell
FISH AND TRANSLOCATIONS
Cells which contain balanced translocations
tend not to be eliminated at cell division
Difficult to see in a conventional preparation,
but easy to observe with FISH
 Probes now available for every human
chromosome that make them fluorescent in
bright colours
Since this is a stable aberration, the
advantage of this method is its application to
persons exposed to radiation at some time in
the past

Chromosome Aberrations

Uploaded by

Copyright:

Available Formats

Chromosome Aberrations

Uploaded by

Document Information

Original Description:

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Chromosome Aberrations

Uploaded by

Copyright:

Available Formats

CHROMOSOME

From: Brum GD, et al (1989). Biology:Exploring Life pg. 156

Dicentric (or Tricentric)

From: Radiobiology for the Radiologist, pg 24

 The sticky ends may rejoin to form a ring with a

 Later during DNA synthetic phase the

From: Radiobiology for the Radiologist, pg 24

From: Radiobiology for the Radiologist, pg 24

persons (about 1/1000 cells)

common as excess fragments

 Have a higher occurrence (1/300 cell) in

You might also like