Cytogenetics

Lecture 1

Faculty of Health and Wellness Sciences

Department of Biomedical Sciences
BHSc: Medical Laboratory Science
Mrs R Jacobs Snyders
2024
 Role of Cytogenetics in the medical laboratory

Cytogenetics

The analysis of various human tissue cells that reveals

the organization of chromosomes - morphology,
structure, pathology, function, and behaviour

Chromosomes

The physical structures that contain the genetic

material, DNA.
 Role of Cytogenetics in the medical laboratory

Samples brought to the Cytogenetics lab are studied:

To aid in the diagnosis of inherited conditions –

constitutional

&

To diagnose or monitor cancerous conditions –

acquired with associated chromosome abnormalities
 History

Time line

 1952 “dark ages” 2n=48

 1950’s Use of PHA (phytohemagglutinin) &
hypotonic (low salt) solution
 1956 2n=46
 1959 “trisomy period” extra 21 chr
 1960’s “banding era” – differential staining

“Molecular era” – Higher resolution screening.

 The Cell Cycle

Two modes of cell division:

Mitosis  responsible for proliferation of body/somatic

cells

Meiosis  responsible for production of gametes

The Cell Cycle
The Cell Cycle
 Chromosomes

Diploid nr of chromosomes  in autosomes

Haploid nr of chromosomes  in gametes

Chromosome Shape
 Chromosome Shape

 Replicated chromosomes at metaphase consist of

sister chromatids joined by a single centromere

 As chromosomes condense and become visible

during cell division, i.e. at Metaphase certain
features can be recognized

Centromere

A region of a chromosome to which microtubule

fibres attach during cell division & the location of a
centromere gives a chromosome its characteristic
shape
 Metaphase Chromosomes

Chromosomes are identified by the following:

 Size (from largest to smallest)

 Centromere location (metacentric, sub-

metacentric & acrocentric)

 Banding pattern (G-banding)

Metaphase Chromosomes
 Metaphase Chromosomes

15
8 Acrocentric
1
Submetacentric
Metacentric
 Types of Chromosomes

Sex Chromosomes

 X and Y chromosomes that are involved in sex

determination
 Different sizes and shapes

Autosomes

 Chromosomes other than the sex chromosomes

 In humans – chromosome 1 - 22
 How are Human Chromosomes analysed?

Karyotype

A complete set of chromosomes from a cell that has

been photographed during cell division at the
metaphase stage and arranged in a standard
sequence.
How are Human Chromosomes analysed?

Metaphase
Chromosomes arranged in a karyotype
 Chromosomes arranged in a karyotype

46,XX
Normal female
karyotype
Staining Procedures

 Routine stain in
cytogenetics

 Used to identify
Y chromosome

 Useful for examining

telomeres

 Used to identify
centromeres in
Dicentric chromosomes
 Staining Procedures

Quinacrine  Y chromosome, acrocentric long satellites

vs. Extra material
 Staining Procedures

C- banding  variations of heterochromatic regions,

dicentric chromosomes
 Staining Procedures

NOR (nucleolus organizing regions) Silver Staining 

studying satellites and stalks of acrocentric chromosomes
 Basic procedure
Specimen reception Culturing

Analysing
Reporting

Harvesting
Staining

Slide making
Basic procedure

Automatic Karyotyper
 Information obtained from a Karyotype

 Number of chromosomes:
Presence/absence of individual
chromosomes  Numerical disorders

 Sex chromosome content:

Identify & confirm sexual phenotype.
Distinguish between autosomes & sex
chromosomes.

 Structural Abnormalities:
Resulting from deletions, insertions,
duplications, translocations, and
environmental factors.
Karyogram: Chromosome Banding Patterns

• Normal chromosomes share

a common banding pattern

• Abnormalities are detected

when changes are seen in
banding pattern

• ISCN
ISCN

International System
for Human
Cytogenomic
Nomenclature
2020
 ISCN

 Allows any region to be identified by a

descriptive address (chromosome nr,
arm, region, and band)

Karyotype is written according to:

• Chromosome number
• Sex chromosomes
• Abnormality located using p and
q band numbering

Example:

46,XY,del(1)(q24)
ISCN – Chromosome grouping
ISCN
 ISCN

• Identification of a chromosome
by length and position of
centromere.
• Accurate reporting of
abnormalities.
• The smaller the defect, the
longer the chromosome needs
to be for detection – banding
quality
• Lower banding is adequate for
NUMERICAL abnormalities e.g.
400 bands.
• Higher banding is necessary for
structural abnormalities such as
deletions, translocations and
inversions.
300 400 550 850
Microscopic to Molecular
Microscopic to Molecular
Reasons for Cytogenetic Referrals
Chromosomal Abnormalities in Miscarriages
 Chromosomal Abnormalities

Aneuploidy  involves the gain or loss of individual

chromosomes

Monosomy  a condition in which one member of a

chromosomal pair
is missing; one less than diploid number (2n-1)
e.g. Turner Syndrome

Trisomy  a condition in which one chromosome is present in

three copies,
and all others are diploid; one more than the
diploid number (2n+1)
e.g. Down syndrome
 Causes of Aneuploidy

Nondisjunction

The failure of homologous chromosomes to

separate properly during either meiosis I or
meiosis II
 Nondisjunction

Meiosis  formation of gametes

Gametes  contain the haploid (half) nr of

chromosomes

+ = zygote

Nondisjunction can lead to an error in chromosome

number resulting in an abnormal clinical phenotype
 Nondisjunction

 May be maternal or paternal origin

 In most cases, the origin is maternal, where the
mother conceives at an older age.
 Due to older age, the maternal oocyte is exposed to
harmful environmental influences for a longer period.
 Other predisposing factors are radiation, viral
infections and genetic predispositions.
 The sperm has a shorter lifespan and therefore has
less chances of injurious exposure.
 Why is Maternal Age a Risk Factor?

Meiosis is not completed until ovulation

 Intracellular events may increase
risk of nondisjunction, resulting
in aneuploidy

Maternal selection
 Embryo-uterine interactions that
normally abort abnormal embryos
become less effective

Age of the mother is the best known risk factor for

trisomy
Why is Maternal Age a Risk Factor?
Meiosis
Meiosis

Females:
Delay in
completion of
the meiotic
cycle
Nondisjunction during meiosis
Nondisjunction during meiosis I
Nondisjunction during meiosis II
 Effects of Monosomy & Trisomy

Autosomal monosomy

 Lethal condition
 Eliminated early in development

Some autosomal trisomies

 Relatively common
 Most result in spontaneous abortion
 Three most common types that can result in live
births (13, 18, 21)
Trisomies in Spontaneous Abortion
 Trisomy 13 – Patau Syndrome

47,XX,+13
47,XY,+13

 Lethal condition
 1 in 10 000-20 000 births
 Most die within 1st month

Clinical features:

 Polydactyly
 Eye defects
 Severe brain, nervous system &
heart defects
Trisomy 13 – Karyotype

47,XX,+13
 Trisomy 18 – Edward Syndrome

47,XX,+18
47,XY,+18

 Lethal condition
 1 in 6 000- 8000
births
 Survival only 2-4
months
 Trisomy 18 – Edward Syndrome

Clinical features:

 IUGR
 Dolicocephaly
 Prominent occiput
 Short nose; upturned nares
 Small mouth – cleft palate may be present
 Low set, dysmorphic, posteriorly rotated auricles
 Short neck
 Clenched fists; overlapping fingers
 Dislocation of hip
 Rocker bottom feet
 Trisomy 21 – Down Syndrome
47,XX,+21
47,XY,+21
 Occurs in 1/700 births
 Only autosomal trisomy that
allows survival into adulthood

Clinical features:

 Mental Retardation
 Epicanthic fold (corner of eye)
 Large furrowed tongues
 40% have congenital heart
defects
 High risk of leukaemia &
Alzheimer’s disease
 Few reach age of 50
Trisomy 21 – Karyotype

47,XX,+21
 Aneuploidy of the Sex Chromosomes

 More common than autosomal aneuploidy

 Can involve both X and Y chromosomes
 A balance is needed for normal development
 At least one copy of the
X chromosome is required for
development
 Increasing numbers of X or Y
chromosomes causes progressively
greater disturbances in phenotype &
behaviour
 Turner Syndrome

 Monosomy of the X chromosome  results in female sterility

 Other phenotypic characteristics but otherwise normal.

45,X
 Klinefelter Syndrome

 Males have some fertility problems but few

additional symptoms

47,XXY
 XYY Syndrome
 Affected individuals are usually taller than normal and some,
but not all, have personality disorders
 Aneuploidy of the Sex Chromosomes

Changes in the number of

sex chromosomes have less
impact than changes in
autosomes