KARYOTYPING

Karyotype • A karyotype is the number and appe

arance of chromosomes in the nucleu
s of a eukaryotic cell.

Analyzes:
• Sex chromosome content
• Presence or absence ofindividual chro
mosomes
• Nature or extent of chromosomal aberr
ations

The term is also used for the complete set of ch

romosomes in a species, or an individual organi
sm.
Karyotype • Karyotypes describe the number of chromosomes,
and what they look like under a light microscope.
• Attention is paid to their length, the position of the c
entromeres, banding pattern, any differences betwe
en the sex chromosomes, and any other physicalchar
acteristics.
• The preparation and study of karyotypes is part o
f cytogenetics.
• Genetic counseling is a relatively new field that allow
s parents the opportunity to determine the genetic
odds that their offspring may carry a particular negati
ve (or lethal) trait.
• As technology improved, a process called karyotype
analysis was developed. This procedure provides inf
ormation regarding the entire chromosomal compl
ement of an individual, as viewed during the proces
s of mitosis.
 Karyotyping Analysis
• Karyotypes may be performed by taking (1) a blood, bone marrow, or skin
sample from an adult; or (2) a sample of amniotic fluid (which contains stray
cells) or extra-embryonic cells (from the chorionic villi) from an unborn child.
• Once the cells have been obtained for a karyotype, they are mixed with plant-
derived chemicals called lectins that stimulate mitosis.
• After the required quantity of cells has been acquired, the cells are treated
with a drug called colchicine to stop the mitotic process at metaphase. Col
chicine arrests the action of spindle fiber microtubules; hence ceasing the m
itotic process.
• The cells are placed in a hypotonic solution (see: osmosis and diffusion lab)
that causes them to take on water, thereby increasing their overall size.
• This provides room for chromosomes to be spread out. The cells are fixed to a
microscope slide, stained and then photographed.
 • After they are photographed, the photo is enl
arged and the chromosomes are cut out and m
Karyotypin g •
atched up aspairs.

Analysis
Chromosomes may be distinguished from one
another based upon several keycharacteristics:
• (1) the length of the arms of the chromosome; a
pattern that is established by the position of the
• centromere along thearms,
• (2) shape and
• (3) general appearance of the chromosome, such
as size and placement of bands.

Karyotype analysis provides a mechanism to determ

ine if nondisjunctionaldiseases, such as Down
syndrome (Trisomy 21), are present in the fetus.

It also provides a look at the chromosomes to see if

there are any missing segments (deletions) or tran
slocations that may have occurred during crossin
g-over.
Karyotype
NORMALCELL CANCERCELLS
Karyotype • The study of whole sets of chromosomes is
sometimes known askaryology.
• The chromosomes are depicted (by rearranging a m
icrophotograph) in astandard format known as a ka
ryogram or idiogram:
• in pairs, ordered by size and position of
centromere for chromosomes of the same size.
• The basic number of chromosomesin
the somatic cells of an individual or a species is called
the somatic number and is designated 2n. Thus, in hu
mans 2n = 46. In the germ-line (the sex cells) the chr
omosomenumber
is n(humans: n = 23).
• So, in
normal diploid organisms, autosomal chromos omes
are present in two copies. There may, or may not, be
sex chromosomes.
• Polyploid cells have multiple copies of chromoso
mes and haploid cells havesingle copies.
 • Polyploidy – a chromosome number t
hat is a multiple of the normal haploid
set
•Aneuploidy – a chromosomalnumber
that varies by something less than a set

• –Monosomy – having only one member

Variations in of a homologouspair
Chromosome N • –Trisomy – having three copies of a
umber single chromosome
 • It is estimatedthat
• Humans have a rate of aneuploidy 10x high
er than other mammals, including primate
s
•1 in 2 conceptions areaneuploid
•35–70% of early embryonic deaths and spo
ntaneous abortions are caused by aneuploi
Aneuploidy Is dy
a Major Cause • 1in 170live births are at least partially an
of Reprod euploid

uctive Failure • 5–7%of early childhood deaths are rel

ated to aneuploidy
• Polyploidy
• Caused by
• Errors in meiosis
• Events at fertilization
• Errors in mitosis

• Triploidy
• Three sets of chromosomes (69)
• Most common form of polyploidy
• 15–18% ofall spontaneous abortions
• Approximately 75%have two sets of paternal chromosomes
• Probably due to polyspermy
• 1%conceptions are triploid but 99% die before birth (lethal condition)
Tetraploidy
• Four sets ofchromosomes (92)
• 5% of allspontaneous abortions
• Extremely uncommon in live births
• May result from failure of cytokinesis in the 1st mitotic
division
• Life threatening
 Most Common Cause of Aneuploidy
Is Nondisjunction in Meiosis
Nondisjunction is the failure of homologs
or sister chromatids chromosomes to
separate in meiosis or mitosis

• Produces abnormal gametes

• Phenotypic effects of aneuploidy vary
widely
Nondisjunc
tion
Autosomal • • Lethal condition
Monosomy • • Aneuploidy during gamete
formation produces equal
numbers ofmonosomic and
trisomic gametes and emb
ryos
• • Rarely seen in spontane
ous abortions and live birt
hs
• • Majority are lost early in
development
Autosomal • Most arelethal
• 50% of cases of chromosomal
Trisomy abnormalities that cause fetal
death are autosomaltrisomies
• Varies by chromosome
• Examples
• Trisomy 13: Patausyndrome
(47,+13)
• Trisomy 18: Edwardssyndrome
(47,+18)
• Trisomy 21: Downsyndrome
(47,+21)
Trisomy 18: • 1/11,000 births
Edwards • Average survival time2–4
Syndrome months
(47,+18) • Affected infants small at bi
rth grow slowly and are me
ntally retarded
• Malformation of heart, h
ands, and feet
• For unknown reasons 80%of
all trisomy18 are female
• Advanced maternalage is a ri
sk factor
Edward’s
Syndrome
Trisomy 13:
Patau • 1/15,000 births
Syndrome • Lethal; mean survival time 1
month
(47,+13
• Facial malformations, eye de
fects, extra fingers or toes, a
nd large protrudingheels
• Severe malformations of brain,
nervous system, andheart
• Parental age only known risk
factor
 Patau’s
Syndrome
Trisomy 21:
• First chromosomal abnor
Down mality discoveredin hum
Syndrome ans (1959)
(47,+21) • 1/900 live births
• Leading cause of mental
retardation and heart d
efects in US
• Wide flat skulls, skin folds in
the corner of the eyes, spots
on the irises, and thick, furr
owed tongues
• 40% congenital heart
defects
 Down
Syndrome
 Risk for
Autosomal
Trisomy

• Advanced maternal age

• Risk increases rapidly after35
years of age
 Turner’s • 1/10,000 births
• Females; short, wide chest;rudimentary
Syndrome: ovaries; and abnormal sexual develop
45, X ment
• Puffiness ofhands and feet
• Abnormalities ofthe aorta
• No mentaldysfunction
• Single X chromosome; two X chromos
omes are required fornormal female s
exualdevelopment
• Complete absence of an X chromosome
is lethal- (So, no Ymonosomies)
 Turner’s
Syndrome
Klinefelter • 1/1000
• Features do not develop until after
Syndrome: puberty
47, XXY • Affected individuals are male with low
fertility and may have mental dysfun
ction
• 60% due tomaternal nondisjunction
• Other forms XXYY, XXXY and XXXXY
Klinefelter’s
Syndrome
 XYY • 1/1000 births

Syndrome: • Above average inheight

• No established link with possible
47, XYY antisocial behavior