CYTOGENETICS

Chapter 6

A. Cytogenetics
- Is a discipline within genetics
- Deals with chromosome variations
- In general, excess genetic material has milder effects on health than a deficit
- Still, most large-scale chromosomal abnormalities present in all cells disrupt or halt prenatal
development
B. Portrait of a Chromosome
- A chromosome consists primarily of DNA and protein
- Distinguished by size and shape
- Essential parts are:
. Telomeres
. Origins of replication sites
. Centromere

Heterochromatin - darkly staining

- Consists mostly of repetitive DNA
Euchromatin - Lighter Staining
- Contains most protein-encoding genes
Telomeres - Are the chromosomes tips
- Composed of many repeats of TTAGGG
- Shorten with each cell division

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C. Karyotype
- A chromosome chart
- Displays chromosomes arranged by size and structure
- Humans have 24 chromosome types:
Autosomes are numbered 1-22 by size
Sex chromosomes are X and Y

Karyotype analysis can reveal chromosome abnormalities:

- Changes in chromosome number (too many or too few is bad)

- Changes in chromosome structure (too many or too few genes is also bad)
i. Normal human chromosome number is 46 with 44 autosomes and two sets of sex
chromosomes
ii. Normal human development requires 2 copies of most genes, if 2 copies are not
present there are “GENE Dosage” problems. Developmental abnormalities and
lethality are typical results of gene dosage problems
iii. Visualizing Chromosomes:
- Tissue is obtained from person:
Fetal tissue: Amniocentesis
Chronic villi sampling
Fetal cell sorting
Chromosome microarray analysis
Adult tissue: White blood cells
Skin-like cells from cheek swab
- Chromosomes are extracted
- Then stained with a combination of dyes and DNA probes
D. Prenatal Diagnosis often uses karyotype examinations
I. Reasons for doing prenatal diagnosis include:
. Maternal age > 35

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. Previous chromosomally abnormal child recurrence risks usually approach 1-2 %
. One parent carries balanced translocation
. Woman is heterozygous for x-linked recessive gene
. Both parents are heterozygous for autosomal recessive mutation in a known location
on the chromosomes
II. Maternal Age and Trisomic Conceptions

Visualizing chromosome by fetal tissue:

Amniocentesis:
- Detects about 1,00 of each of the more than 5,000 known chromosomal and biochemical
problems – culture for 10 days – probes detect chromosomes in 1-2 days
- Ultrasound is used to follow needle’s movement

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Chorionic Villi sampling:
- Performed during 8th week of pregnancy-use cells directly for testing
- Provides earlier results than amniocentesis
- However, it does not detect metabolic problems
- And has greater risk of spontaneous abortion
- Less accurate than amniocentesis

Fetal Cell Sorting:

- Fetal cells are distinguished from maternal cells by a fluorescence-activated cell sorter –
Identifies cell - surface makers -70% of pregnancies
- Considered to be a new technique of detecting fetal mRNA in the blood stream of the
mother

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Chromosome microarray analysis:

Drawing by German biologist Walther Fleming Micrograph of actual stained human chromosomes
1882 Now

FISH:
- Fluorescence in situ hybridization
- DNA probes labeled with fluorescing dye bind complementary DNA
- It is used to detect the physical location of a gene on an intact chromosome.
- “in situ” is Latin for “in place” and in this method the chromosomes are adhered “in place”
on a slide

Fluorescent dots
correspond to 3 copies of
chromosome 21

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There are 2 classes of chromosome number abnormalities:

A. Polyploidy – extra complete sets of chromosomes 3n-triploidy, 4n-tetraploidy
***Polyploidy is not compatible with human life-fetuses are aborted spontaneously or infants
die shortly after birth***
1. Polyploids are common among plants, bananas and seedless watermelons are examples of
triploids (triploids have problems producing ‘balanced gametes’ and are sterile as a result).
Bread wheat is a hexaploidy.
2. Caused by
Errors in meiosis
Events at Fertilization
Errors in mitosis
3. Examples:

Triploid (3n):

- Three sets of chromosomes (69 chromosomes)

- Most common form of human polyploidy 15-18% of all spontaneous abortions
- Approx. 75% have 2 sets of paternal chromosomes, due to polyspermy (Dispermy)
- 1% conceptions are triploid, but 99% die before birth

Tetraploid (4n):

- 4 sets of chromosomes (92), 5% of all spontaneous abortions

- Extremely uncommon in live births
- Most result from failure of cell division in the 1st mitotic division of the zygote
B. Aneuploidy:
Failure of chromosomes separation (nondisjunction) in Meiosis I or of chromatid separation in
meiosis II produces gametes with 22 or 24 chromosomes, only one chromosome pair is usually
involved
a. Monosomy-zygote with 45 chromosomes
2n-1 chromosome
b. Trisomy-zygote with 47 chromosomes
2n+1 chromosome
c. Aneuploidy is a major cause of human reproductive failure, it is estimated that:
– Humans have a rate of aneuploidy 10x higher than other mammals, including primates
– 1 in 170 live births are at least partially aneuploidy
The most common cause of aneuploidy is nondisjunction occurring in meiosis in females.
***NONDISJUNCTION is the failure of homologs or sister chromatids to separate in meiosis or
mitosis, it leads to ‘unbalanced’ gametes with too many or too few genes on the chromosomes
involved***

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Nondisjunction in Meiosis I for a Nondisjunction in Meiosis II for a

female (XX) female (XX)

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1. Autosomal Monosomy (2n-1); Often a lethal condition
A. Majority are lost very early in development
2. Autosomal Trisomy (2n+1)
A. The frequency and severity of the abnormality varies by chromosome

Aneuploidy Trisomy 21

Means that there are 3

copies of chromosome
21

Autosomal Trisomy

Shows the frequencies

of trisomy of different
chromosomes

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Examples of Human Autosomal Aneuploidy

1. Trisomy-21 (Down syndrome), severe – but variable effects on lifespan and other symptoms,
most fetuses with trisomy 21 are aborted but there is a relatively high frequency of this
condition among live births - 1 in 900. It is the leading cause of mental retardation and
congenital heart defects in USA.
Advanced maternal age is a risk factor for having children
with these trisomies.
Why is Age a risk? Because the homologous chromosomes have been sticking to each other for
a long time and they may not go apart from each other

Down syndrome includes a combination of birth defects:

- Most common chromosomal defect in humans

- including some degree of mental retardation
- characteristic facial features
- 40% have some kind of heart defects
- visual and hearing impairment and other health problems.
The severity of all of these problems varies greatly among affected individuals.

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2. Patau Syndrome (Trisomy 13):
- A newborn male with full trisomy 13 (Patau syndrome).
- This baby has:
• a cleft palate
• atrial septal defect
• inguinal hernia
• postaxial polydactyl of the left hand.

3. Edwards Syndrome: trisomy 18; 47 +18

It is a severe chromosome abnormality where the child has an extra chromosome 18 in every cell.
There are three types of the syndrome:

Full form (severe) - in this every cell in the body has three chromosome 18's instead of two.

Mosaic form (less severe) - in this some cells have two chromosome 18s while others have three.
The extent and severity of the condition will depend upon the ratio of normal to abnormal cells.

Partial form - in some cases there may be an extra copy of part of chromosome 18. This is referred
to as partial trisomy 18. The effects of this may be milder and would require further medical advice.

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Children with all their cells affected do not normally survive beyond infancy. Those affected by the
mosaic and partial forms may survive to adulthood.

Trisomy 18 is a relatively common syndrome affecting approximately 1 out of 11,000 live births. It is
three times more common in girls than boys. The extra material interferes with normal development.

Add to notes …

Medical diagnosis:

 Clenched hands

 Crossed legs (preferred position)

 Heart disease (congenital)

 Mental deficiency

 Grow very slowly

Human Sex Chromosome Aneuploids:

A. Monosomy - XO (Turner’s syndrome or Ullrich Turner) - 45, XO female, sexual
a. development problems, sterile, very common in zygotes but nearly all aborted, rare in live births
- 1 in 10,000 female live births, no effect on lifespan or mental function
b. (XO) encompasses several chromosomal abnormalities, of which monosomy X is the most
common.
c. It occurs in about 1 out of every 2500 female births.
d. individuals with Turner syndrome are 45,X. In Turner syndrome, female sexual characteristics
are present but generally underdeveloped.
e. short stature, webbed skin of the neck, abnormal bone development, such as a "shield-shaped,“
broad flat chest ,absent or retarded development of secondary sexual characteristics that
normally appear at puberty, infertility, absence of menstruation
B. Trisomy - XXY (Klinefelter syndrome) - 47, XXY male, sexual
a. development problems at puberty, low fertility, common in zygotes and live births - 1 in 1,000
male births, not aborted – no effect on lifespan, often some effect on mental function
b. is a condition caused by a chromosome aneuploidy.
c. Affected males have an extra X sex chromosome. The principal effect is small testes
development and reduced fertility.
d. A variety of other physical and behavioral differences and problems are common, though
severity varies and many boys and men with the condition have few detectable symptoms.
e. It is the second most common extra chromosome condition, and is named after Dr. Harry
Klinefelter, an endocrinologist who first described it in 1942. The condition exists in roughly 1
out of every 500 to 1,000 males.

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C. Trisomy XYY (Jacob’s syndrome) - 47, XYY male,

a. normal fertility and sexual development, common - at least 1 in 1,000 male births, not
aborted, above average in height, at most minor effects on mental function and social
behavior
b. As adults, these "super-males" are usually tall (above 6 feet) and generally appear and act
normal.
c. However, they produce high levels of testosterone. During adolescence, they often are
slender, have severe facial acne, and are poorly coordinated.

D. Trisomy XXX (Super female syndrome) - 47, XXX,

- no problems, at least 1 in 1,000 female births, not aborted
- Generally are an inch or so taller than average with unusually long legs and slender torsos, but
otherwise appear normal.
- They usually have normal development of sexual characteristics and are fertile.
- They may have slight learning difficulties and are usually in the low range of normal intelligence.

Barr Bodies Dosage Compensation:

Dosage compensation
Only one X chromosome will be activated in each cell; the other will form a Barr body (coiled chromatin)
in a female. Males only have one Barr body.
No Barr Bodies in Males; only one in each female

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A. Prader-Willi Syndrome:
i. Infants with partial deletion of chromosome 15
ii. Weak slow growth
iii. Children and adults display obesity and compulsive eating

B. Fragile X Syndrome:
i. Associated with a mental retardation known as Martin-bell syndrome
ii. Lower portion of the X chromosome appears to be hanging by a thread
iii. Extended jaw and large ears; degree of mental retardation
iv. Mom can be a carrier; most often found in males

Fragile Sites on the Human X chromosome:

- The FRAX A site is responsible for Fragile X Syndrome

- It is near the end of the long arm of the X chromosome as in picture
C. Cri Du Chat Syndrome
i. Caused by a deletion of chromosome number 5
ii. Child is mentally retarded; with defects in facial development, the glottis and larynx, and cry
sound like a cat
D. Deletions
A deletion refers to a missing genetic segment from a chromosome
Deletions are often not inherited
- Rather they arise de novo
Larger deletions increase the likelihood that there will be an associated phenotype
Cri-du-chat (cat cry) syndrome
- Deletion 5p–

2 Types of Translocation
Reciprocal Translocation Robertsonian Translocation

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2 nonhomologous chromosomes exchange parts Centromeres from two nonhomologous
chromosomes fuse and chromosomal material
from the short arms is lost.
No genetic information is gained or lost except 5% of Down syndrome cases involve a
for possibly a pair of genes affected at the Robersonian translocation between
breakpoints chromosomes 21 and 14.
If a parent is a CARRIER of a 14/21 chromosome
there is a 1 in 3 risks of a Down syndrome child,
typically down syndrome (true aneuploidy)
doesn’t run in the family.
Translocation Down syndrome does run in the
families, it is an inherited condition

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