Experiment 1

Title: Study of chromosomes in cell division

Introduction:

The passing down of genetic instructions from one generation to the next generation is called
inheritance. The genetic material, the DNA is carried in the chromosomes. When
homologous chromosomes are arranged from the largest pair to the smallest pair and
numbered according to their size, they form the karyotype of an individual. The cells of an
individual with a genetic disease or genetic disorder show a different karyotype from that of
a normal human being.

Aim: To prepare a karyotype and subsequently identify the phenotype of the individual

Materials:

Scissors, glue, photocopied randomly arranged chromosomes paper (figure 2), and A4 paper.

Procedure:

1. Figure 1 showed the karyotype (chromosome in pairs) of a normal human male while
figure 2 showed the karyotype of a human with the chromosomes randomly arranged.
2. Using a pair of scissors and Figure 1 as reference, the individual chromosomes from
figure 2 are cut out, arranged in pairs, and stuck on a piece of A4 paper which is to be
submitted together with the lab report.
3. Whether the karyotype belongs to a normal or abnormal male or female is identified.
If it belongs to an abnormal person, the abnormality is identified.

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Discussion:

Every cell has thread-like structures in its nucleus called chromosomes. A chromosome is an
organised structure of DNA and protein that is found in cells. Each chromosome is made up
of DNA tightly coiled many times around proteins called histones that support its structure.
Without such packaging, DNA molecules would be too long to fit inside cells. A
chromosome contains many genes, regulatory elements, and other nucleotide sequences.
Each chromosome has a constriction point called the centromere, which divides the
chromosome into two sections. The location of the centromere on each chromosome gives the
chromosome its characteristic shape, and can be used to help describe the location of specific
genes. The number of chromosomes present in the cells of each species of an individual
organism is constant. This number is referred to as the chromosomal number of the species.
Cells of individuals of a different species have a different chromosomal number. For
example, onions have 16 chromosomes, dogs have 78, while the fruit fly, Drosophila
melanogaster has 8 chromosomes. Humans, on the other hand have 46 chromosomes. The
nucleus of most human cells contains two sets of chromosomes, one set given by each parent.
Each set has 23 single chromosomes – 22 autosomes and an X or Y sex chromosome. (A
normal female will have a pair of X chromosomes; a male will have an X and Y pair.)
Chromosomes contain roughly equal parts of protein and DNA. DNA molecules are among
the largest molecules now known. Chromosomes are not visible in the cell’s nucleus – not
even under a microscope, when the cell is not dividing. However, the DNA that makes up
chromosomes becomes more tightly packed during cell division and is then visible under a
microscope. The DNA becomes supercoiled and condensed to facilitate distribution into
daughter cells. At this time, chromosomes can be seen under a light microscope and, when
stained with certain dyes, reveal a pattern of light and dark bands reflecting regional
variations in the amounts of A and T versus G and C. Differences in size and banding pattern
allow the 24 chromosomes to be distinguished from each other, an analysis called a
karyotype.

In relation to that, karyotype refers to the chromosomal complement of an individual,

including the number of chromosomes and any abnormalities. The term is also used to refer
to a photograph of an individual's chromosomes. The preparation and study of karyotypes is
part of cytogenetics. Usually, metaphase cells are required to prepare a standard karyotype,
and virtually any population of dividing cells could be used. Blood is easily the most
frequently sampled tissue, but at times, karyotypes are prepared from cultured skin fibroblasts

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or bone marrow cells. Karyotype test is a test to identify and evaluate the size, shape, and
number of chromosomes in a sample of body cells. Among the reasons why a karyotype test
would be done is to determine whether the chromosomes of an adult have an abnormality,
whether a chromosome defect is preventing a woman from becoming pregnant or causing
miscarriages, whether a chromosome defect is present in a foetus, and help determine the
appropriate treatment for some types of cancer. In a karyotype test, six different
characteristics of chromosomes are usually observed and compared, namely differences in
absolute sizes of chromosomes, position of centromeres, relative size of chromosomes, basic
number of chromosomes, number and position of satellites, and degree and distribution of
heterochromatic regions. Although the replication and transcription of DNA is highly
standardised in eukaryotes, the same cannot be said for their karyotypes, which are highly
variable between species in chromosome number and in detailed organisation despite being
constructed out of the same macromolecules. Although there are many protocols for
preparing a karyotype from peripheral blood lymphocytes, a rather standard series of steps
are involved. First, a sample of blood is drawn and coagulation is prevented by addition of
heparin. Then, mononuclear cells are purified from the blood by centrifugation through a
dense medium that allows red cells and granulocytes to pellet, but retards the mononuclear
cells (lymphocytes and monocytes). After that, the mononuclear cells are cultured for 3-4
days in the presence of a mitogen like phytohemaglutinin, which stimulates the lymphocytes
to proliferate vigorously. At the end of the culture period, when there is a large population of
dividing cells, the culture is treated with a drug such as colcemid, which disrupts mitotic
spindles and prevents completion of mitosis. This greatly enriches the population of
metaphase cells. The lymphocytes are then harvested and treated briefly with a hypotonic
solution. This makes the nuclei swell osmotically and greatly aids in getting preparations in
which the chromosomes don't lie on top of one another. Following that, the swollen cells are
fixed, dropped onto a microscope slide and dried. Slides are stained after treatment to induce
a banding pattern as described. A digital image of the chromosomes can be cut and pasted
using a computer. If standard staining is used, the orderly arrangement is limited to grouping
like-sized chromosomes together in pairs, whereas if the chromosomes were banded, they can
be unambiguously paired and numbered.

Genetic disorders sometimes occur in humans due to several factors. Down’s syndrome is an
example of a genetic disorder in humans. Down syndrome (DS), also called Trisomy 21 (the
presence of extra genetic material on the 21st pair of chromosomes), is a condition in which

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extra genetic material causes delays in the way a child develops, both mentally and
physically. Therefore, a person has 47 chromosomes instead of the usual 46. It affects about 1
in every 800 babies. The risk is higher with increasing maternal age. The affected child is
born with characteristic physical features and may have several associated medical problems.
Kids with Down syndrome tend to share certain physical features such as a flat facial profile,
an upward slant to the eyes, small ears, and a protruding tongue. Low muscle tone (called
hypotonia) is also characteristic of children with DS, and babies in particular may seem
especially "floppy." Though this can and often does improve over time, most children with
DS typically reach developmental milestones much later than other kids. Down syndrome
affects kids' ability to learn in different ways, but most have mild to moderate intellectual
impairment. Generally, individuals with Down syndrome tend to have a lower than average
cognitive ability, often ranging from mild to moderate developmental disabilities. A small
number have severe to profound mental disability. Kids with Down’s syndrome can and do
learn, and are capable of developing skills throughout their lives. They simply reach goals at
a different pace. Health concerns for individuals with Down syndrome include a higher risk
for congenital heart defects, gastro esophageal reflux disease, recurrent ear infections,
obstructive sleep apnea, and thyroid dysfunctions. Usually, confirmation that a child has
Down’s syndrome is by a blood test on the child to detect the extra chromosome 21. It is also
possible to diagnose the condition during pregnancy with blood tests, detailed
ultrasonography or amniotic fluid analysis.

Conclusion:

The karyotype sorted out from figure two belongs to an abnormal human male. An extra
chromosome on chromosome 21 causes Down’s syndrome.

References:

Websites:

 http://en.wikipedia.org/wiki/Chromosome.
 http://www.webmd.com/baby/karyotype-test.
 http://en.wikipedia.org/wiki/Karyotype.
http://en.wikipedia.org/wiki/Homologous_chromosome.
 http://www.bio-medicine.org/biology-dictionary/Homologous_chromosome/.