What Is a Chromosome?

A long, continuous strand of DNA, plus several

types of associated proteins, and RNA.

 Cytogenetics: the study of chromosomes, their

structure, and their inheritance.
The importance of chromosomes in medicine:
Clinical diagnosis
Eg, Turner syndrome

Gene mapping
Eg, cystic fibrosis

Cancer cytogenetics
Eg, retinoblastoma

Prenatal diagnosis
Eg, Down syndrome

Karyotype:apictureofallthechromosomesarrangedbytype:
Karyotypealsoreferstothestandardchromosomesetofanindividualoraspecies.

METACENTRIC
Short
Arm
(p)

SUBMETACENTRIC

ACROCENTRIC
p
Stalk
q
Satellite
Centromere
p

Long
Arm
(q)
3

21

Stalk

17

21

Size:
chromosome 1 is largest, chromosome 22 smallest
Centromere position:
metacentric, submetacentric, acrocentric
all chromosomes have a shorter arm, p, and a longer arm, q.

 G-banding:
pattern subdivides
each chromosome
arm into regions.

Karyotypesaremadefrommetaphasechromosomes:

1.

Cells (from blood, amniotic fluid, or chorionic villus) are grown in culture.
Mitogens may be required: lymphocytes require phytohemagglutinin

2.
3.
4.
5.
6.
7.

Colcemid stops cells at metaphase.

Hypotonic shock ruptures RBCs, swells lymphocytes.
Cells are fixed in MeOH/HOAc.
Chromosomes are spread on a slide.
Trypsinization and staining with Giemsa reveals G-bands.
The chromosome spread is photographed and arranged by type.

Ideogram of G-banding
pattern at 450-band
stage:
High-resolution bandings of 550-850 bands can
be made from prometaphase chromosomes.

Introduction of molecular
cytogenetics:
DNA in chromosome spreads is denatured and
hybridized to probes:
FISH: fluorescent in situ hybridization
Probes for specific genes or chromosomal regions
Rearrangements, deletions, abnormal chromosome number

Chromosome painting
Mixture of fluorescently-labeled probes for single-copy genes that
map along the entire length allow entire chromosomes to be
visualized
Anueploidies, translocations

SKY: spectral karyotyping

All chromosomes painted at once, each chromosome a different color

FISH : gene-specific probes

Pinkel D, Straume T, Gray JW. Cytogenetic analysis using quantitative, high-sensitivity, fluorescence
hybridization. Proc Natl Acad Sci U S A. 1986;83:2934-2938.

A,Partialmetaphaseshowingamicrodeletionoftheelastingene(ELN)(arrow)associatedwithWilliams
syndrome.
B,Interphasecellshowsamicroduplicationoftheperipheralmyelinprotein22gene(PMP22)(arrows)
associatedwithCharcotMarieToothsyndrome.

SKY: chromosome-specific probes

NormalkaryotypeCancercellkaryotype
note:translocations,polysomy,monosomy

Cells used for karyotyping:

Blood cells:
T-lymphocytes from peripheral blood are easily collected
Growth in culture requires mitogens (phytohemagglutinin)
Cultures are short-lived

Skin cells
Fibroblast cultures grow without mitogens
Cultures are long-lived, can be immortalized

Tumor cells
Obtained from biopsy

Amniocytes
From amniocentesis
Amniotic fluid is available after ~16 weeks
Amniotic fluid is waste containing cells shed from skin, respiratory tract, urinary tract.
Amniotic fluid can be biochemically tested for metabolic disorders

Placental cells
Obtained by chorionic villus sampling
Placental cells are dividing, and karyotypes can be obtained within hours

Indications for chromosome analysis:

Family history can indicate a need for prenatal testing:
Amniocentesis and chorionic villus sampling pose small risks of infection or
inducing spontaneous abortion, so samples are taken only if indicated by a risk
factor.

Known chromosomal abnormality of first-degree relative

Previous child with chromosomal aberration
Recurrence risk ~1-2%

Advanced maternal age: >35 years

Mother carries X-linked disorder
Male offspring would be at risk

 Prenatal testing:
Amniocentesis:Chorionicvillussampling:
Chorionic
villi
Developing
placenta

Ultrasound to monitor
procedure

Bladder

Developing fetus
Uterus
Chorion
Amniotic
cavity
Rectum

Catheter

 Other reasons for chromosome analysis:

Fertility problems: infertility or repeated miscarriage
Chromosomal abnormalities seen in one or both parents at 3-6%
Translocation or inversion

Stillbirth or neonatal death

Chromosomal abnormalities at ~10%
Karyotyping important for generating family history

Problems with a child

Failure to thrive, developmental delay, multiple malformations,
ambiguous genitalia, mental retardation

Neoplasia: all cancers show chromosomal abnormalities

Karyotype can provide diagnostic or prognostic information

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