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: a picture of all the chromosomes arranged by type:
Karyotype also refers to the standard chromosome set of an individual or a species.

METACENTRIC SUBMETACENTRIC ACROCENTRIC

p Stalk
Short q
Satellite
Arm Centromere
p
(p) p
21 Stalk
Long q q
Arm
(q)

3 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.
Karyotypes are made from metaphase chromosomes:

1. Cells (from blood, amniotic fluid, or chorionic villus) are grown in culture.
Mitogens may be required: lymphocytes require phytohemagglutinin
2. Colcemid stops cells at metaphase.
3. Hypotonic shock ruptures RBCs, swells lymphocytes.
4. Cells are fixed in MeOH/HOAc.
5. Chromosomes are spread on a slide.
6. Trypsinization and staining with Giemsa reveals G-bands.
7. 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, Partial metaphase showing a microdeletion of the elastin gene (ELN) (arrow) associated with Williams
syndrome.
B, Interphase cell shows a microduplication of the peripheral myelin protein 22 gene (PMP22) (arrows)
associated with Charcot-Marie-Tooth syndrome.
 SKY: chromosome-specific probes

Normal karyotype Cancer cell karyotype

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: Chorionic villus sampling:
Chorionic
Ultrasound to monitor
villi
procedure
Developing
placenta

Bladder
Developing fetus

Uterus
Chorion
Catheter
Amniotic
cavity
Rectum
 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
Kelainan Kromosom Pada Abortus
Spontan
Kelainan Kromosom Pada Bayi Baru
Lahir