Laboratory Exercise #6

HUMAN KARYOTYPING

Introduction:
Occasionally chromosomal material is lost or rearranged during the formation
of gametes or during cell division of the early embryo. Such changes, primarily the
result of nondisjunction or translocation, are so severe that the pregnancy ends in
miscarriage – or fertilization does not occur at all. It is estimated that one in 156 live
births have some kind of chromosomal abnormality. Some of the abnormalities
associated with chromosome structure and number can be detected by a test called
a karyotype. A karyotype can show prospective parents whether they have certain
abnormalities that could be passed on to their offspring, or it may be used to learn
the cause of a child’s disability. Karyotypes can also reveal the gender of a fetus or
test for certain defects through examination of cells from uterine fluid – a procedure
called amniocentesis – or through sampling of placental membranes. To create a
karyotype, chromosomes from a cell are stained and photographed. The photograph
is enlarged and cut up into individual chromosomes. The homologous pairs are
identified and arranged in order by size (with the exception of the sex chromosomes;
these appear last). These tests are typically done on a sample of blood, although
any body cell could be used. The cell must be undergoing mitosis – preferably in
metaphase – so that the chromosomes are replicated, condensed, and visible under
a microscope.

Objectives:

1. To understand what a karyotype is and how it is performed. -understand the

Materials:
The following materials are needed to perform this laboratory experience:
*scissors
*tape
*ruler
*small envelope

Procedure:
The following procedure is utilized to perform this laboratory experience:
1. Using the attached sheets, complete four different karyotypes: One normal male,
One normal female, two different disorders of your choice out of the four. Additional
laboratory time may be granted for work above and beyond the four required
karyotypes.
2. Working slowly and carefully, using scissors cut out the chromosome on one page
labeled “1” and find its’ EXACT match elsewhere on the page (it will not be
numbered). Cut out this chromosome and tape BOTH chromosomes side by side on
a “data page” that has the heading filled out.
3. Continue this procedure until you have matched all chromosomes and taped each
of them in the corresponding place on the data page.
4. If you are caught short of time, use the coin envelope to store any chromosomes
you may have clipped out and not matched. DO NOT CUT OUT ALL
CHROMOSOMES AND THEN ATTEMPT TO MATCH THEM!!! Cut out only one at a
time or you will lose chromosomes.
5. In the event that you have an extra chromosome, DO NOT THROW IT OUT! It is
the chromosome that causes your mutation/disorder and you must match it
correctly. 6. Once your chromosomes are all cut out and included in the karyotypes,
answer the questions and complete the lab.

Questions:

1. What four karyotypes did you choose to complete?

2. How could you determine if your karyotype was male or female?

3. Complete the following table:

Scatter Sheet Number: _____

EXPLANATION OF CHROMOSOME DISORDERS

Cri-du-chat
Babies with the "cry of the cat" syndrome have a cry which sounds like that of a cat
in distress because the infant's larynx is improperly developed. The cause of this
condition is a deletion of about half of the short arm of chromosome number five.
Cri-du-chat babies are severely mentally retarded, have a small cranium, a small
jaw and a moon-shaped face. The incidence of this syndrome is 1/100,000 live
births.
Karyotype: 46XX or 46XY with one chromosome #5 upper arm deletion

Down Syndrome
Trisomy 21, one of the most common causes of mental retardation is due to an extra
chromosome 21. This results in a number of characteristic features, such as short
stature, broad hands, stubby fingers and toes, a wide rounded face, a large
protruding tongue that makes speech difficult and mental retardation. Individuals
with this syndrome have a high incidence of respiratory infections, heart defects
and leukemia. The average risk of having a child with trisomy 21 is 1/750 live
births. Mothers in their early twenties have a risk of 1/1,500 and women over 35
have a risk factor of 1/70, which jumps to 1/25 for women 45 or older.
Karyotype: 47XX or 47XY with 3 of the chromosome #21
Edward's Syndrome
This syndrome, trisomy 18 (an extra chromosome 18), produces severe mental
retardation and a highly characteristic pattern of malformations such as elongated
skull, a very narrow pelvis, rocker bottom feet, malformed heart and a grasping of
the two central fingers by the thumb and little finger. In addition, the ears are often
low set and the mouth and teeth are small. Nearly all babies born with this condition
die in early infancy. The frequency of this syndrome is 1/5,000 live births.
Karyotype: 47XX or 47XY with 3 of the chromosome #18

Jacobs
A chromosome abnormality which is caused by nondisjuction (non-separation) of
the Y chromosome during the second phase of meiosis. This condition only occurs
in men and happens when they have an extra Y chromosome. Men with this
abnormality are tall, develop heavy cases of acne and have low mental ability.
Occurrence is 1/1,000 live male births.
Karyotype: 47XYY (extra sex chromosome - extra Y)

Klinefelter Syndrome
A condition occurring in 1/1,000 male live births. Characteristics associated with
this condition are tall stature, small testicles, developed breasts, sterility and mental
deficiency. Most men with this syndrome appear normal in other ways. This
syndrome only occurs in men and affects the sex chromosomes.
Karyotype: 47XXY or 47XXXY (extra sex chromosomes)

Patau Syndrome
This syndrome (trisomy 13 - extra chromosome 13) causes severely abnormal
cerebral functions and virtually leads to death in early infancy. The baby has very
pronounced clefts of the lip and palate, broad nose, polydactyly (extra fingers and
toes), small cranium and nonfunctional eyes. Heart defects and severe mental
retardation are also part of the clinical picture. The frequency is 1/15,000 live
births.
Karyotype: 47XX or 47XY with 3 of the chromosome 13

Triple X
Superfemale (XXX). Occurs at a frequency of 1/1,000 female live births. No specific
abnormalities are associated with this condition. The vast majority of women who
have this condition are normal mentally, but may have underdeveloped genitalia
and limited fertility. They also may have neuromotor delays. This condition only
occurs in females and affects the sex chromosomes.
Karyotype: 47XXX (affects sex chromosomes - extra X chromosome)

Turner Syndrome
This condition happens when an X-carrying sperm fertilizes an ovum that lacks an
X, or when a sperm lacking an X or Y chromosome fertilizes an X-bearing egg. This
syndrome only affects girls and causes them to be missing an X chromosome (XC).
These girls appear to be normal before puberty, although they are shorter and have
a chunky build. At birth, the distinguishable characteristics include a webbed neck.
At sexual maturity, the secondary sex characteristics are not developed. There also
is no menstruation or breast development and they are usually sterile. The
frequency is 1/2,500 live female births.
Karyotype: 45X or 45XO (affects sex chromosomes - missing an X)