[TRANS] LESSON 2: INTRODUCTION TO CYTOLOGY

DOWN SYNDROME (TRISOMY 21)

OUTLINE
I Cytogenetics
A Walther Flemming
i Down Syndrome (Trisomy 21)
II Karyotyping
III Fluorescent In-Situ Hybridization (FISH)
IV DNA Microarray Analysis
V Chromosomes
A Polyploidy
B Homologous Chromosome
C Note: Chromosome
VI Borrelia Burgdorferi
VII Cellular Division
A Gap 0 or G0 Phase
B Interphase
C 3 Stages of Cell Division
D M Phases (Mitosis Phase)
i 2 Stages
• The other term for down syndrome is Trisomy 21.
E Stages of Mitosis • Why trisomy? Tri means three (3) meaning it have 3
i Prophase chromosomes in 21 or extra chromosomes in number 21.
ii Metaphase o Must remember: In each chromosome we only have
iii Anaphase 2 (pair), but in 21 chromosomes it has three (3) and
iv Telophase that’s the reason of having down syndrome.
F Meiosis • The image above is arranged from chromosomes 21-23
G Chiasmata
VIII Substages of Prophase
and it is called Karyotype or Karyotyping which is very
IX Spindle Fibers useful to visualize the structure of chromosomes from
X Important Note chromosomes 21 to chromosomes 23.
o The scientist based to the sizes of the chromosomes,
and if you are going to look from the largest which is
CYTOGENETICS chromosomes 1 and the smallest is chromosomes 22.
• A branch of genetics that studies the function of the cell,
specifically, chromosomes, in the process of inheritance. KARYOTYPING
o We only have 23 pairs of chromosome. • One of the most basic techniques in genetics.
o 22 autosomal or body chromosomes • Refers to the routine analysis of chromosomes at the
o 1 sex chromosomes. metaphase stage.
o Male: XY o Karyotyping is very useful to see if it has a defect. In
o Female: XX other word, useful to detect the inherited from the
o Autosomal or body chromosomes are same. parents and detection of gender. Each chromosomes
have their function in our body.
WALTHER FLAMMING o In what stage in mitosis performed the
• Describe the chromosomes using salamander. Karyotyping? It happens in metaphase, because in
• The number of human chromosomes was mistakenly metaphase all the chromosomes aligned in the cells.
identified to 48. o When we performing karyotyping, it can be using the
o They are mistakenly identified, but with the help of stain (Giemsa or Leishman)
Walther flaming, we described the chromosomes is
only 46.
o Must remember: 46 total of chromosomes, 23 pairs of
chromosomes, 22 autosomal and 1 sex chromosomes.
• In 1956, the date when proved that the chromosomes is
only 46 not 48.

• In the image above, there is no Y in the chromosomes, so

the probability is it pertaining to a female because there is
2 X chromosomes present in the karyotype. Also, it is good

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 TRANS: INTRODUCTION TO CYTOLOGY
to use to visualize or to see if the new born is female or
male.
o MedTech can also perform karyotyping but it
undergoes trainings and seminar to perform
karyotyping.
FLUORESCENT IN-SITU HYBRIDIZATION (FISH)
• Refers to the cytogenetic technique that uses fluorescent
probes that attach to specific chromosome areas with high
sequence complementary.
o What FISH do? It uses probes which is very specific.
Meaning, it attached to the chromosomes that has a
defect. FISH is more expensive because it uses probes
and it performed (FISH) because we want to check if
the new born has abnormalities.
o Fastidious organism is an organism that needs a
special requirement (like nutritional value) for grow.
o Hemophilus influenzae or Influenza they are bacteria
that require V factor and X factor for survival if the
nutrient is not present, they will not live. That is why
they are fastidious; they require special requirement
like nutrition and vitamins to live.
DNA MICROARRAY ANALYSIS
• Microarrays can also be used to study the extent to which
certain genes are turned on or off in cells and tissues.
• Today, DNA microarrays are used in clinical diagnostic
tests for some diseases.
o In DNA Microarray Analysis, we only need 1 specimen
which is the mRNA/messenger RNA.
▪ The messenger RNA has a function to carry
protein information from the DNA in a cell's
nucleus to the cell's cytoplasm (watery interior),
where the protein-making machinery reads the
mRNA sequence and translates each three-base
codon into its corresponding amino acid in a
growing protein chain. Meaning, they carry the
code and the message if the message will undergo
the translation process they are converted into
proteins.
C.J. ANIS, R. ARANGCO, C.D. GECALE, M.D.G. GOLEZ, A.J. MANGILIT | BSMT
o The 3 types of RNA: mRNA, tRNA, and rRNA.
• This cancer cells and normal cells we need to get the mRNA
via RNA isolation, because this mRNA is very useful to
transcribe the proteins.
• Transcriptase Labeling is a reverse transcriptase of
mRNA to DNA.
o The central dogma of cytogenetics or the 3 basics
steps: DNA Replication which is the DNA will convert
into mRNA and will be converted into proteins or amino
acids.
o The process of converting DNA to mRNA is called
Transcription.
o The process of converting mRNA to protein is called
Translation
• Reverse Transcriptase the mRNA will be converted into
DNA. The reverse transcriptase is an enzyme that reverse
from mRNA into DNA. We need to converted back into DNA
because DNA has the only capability to replicate. Once they
are converted into DNA, they are able to undergo
replication process.
o Must remember: The disease that have a reverse
transcriptase is the HIV (if you have HIV the viral lobe
is very high).
▪ We have what we called the window period
means positive in HIV but in the test you are
negative. The window period is every 6 months.
We need to wait 6 months before the test.
CHROMOSOMES
• Are organized structures containing the DNA of an
organism associated with the structural protein called
histones and other factors that help maintain its integrity
and facilitate its formation and replication.
o The only function of Histone is to prevent the DNA from
being tangled or twisted the DNA, since we all know
that the DNA is double helix structure. Just to reiterate,
the histone protects the DNA from damage or twisted.
o Must remember: The basic parts of DNA is Adenine-
Thymine and Cytosine-Guanine. If the DNA Helicase
come, they are able to break the hydrogen bond easily.
o The Y shape region is called Replication fork
meaning the DNA is already destroyed and it formed Y
shape.
o DNA is a double helix, the upper strand called leading
strand 5’-3’ and the lower strand called lagging
strand 3’-5’
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 TRANS: INTRODUCTION TO CYTOLOGY

NOTE: CHROMOSOME
• Linear chromosomes found among eukaryotes.
o Because the DNA of the eukaryotes are linear where
the prokaryotes, they are circular DNA.
o Prokaryotes - Do not have nucleus (Example:
Bacteria)
o Eukaryotes - Have nucleus
• Prokaryotes have circular DNA and are found in the
nucleoid region.

BORRELIA BURGDORFERI
• Causative agent of Lyme disease.
o This organism causes Lyme disease, we acquire
Table No. 1 Common Name, Genus and Species and Their through the bite of infected ticks specifically the
Chromosome Number Ixodes. If we bitten by the Borrelia it is typically a
COMMON GENUS AND DIPLOID characteristics of skin rash called erythema migrans
NAME SPECIES CHROMOSOMES (The redness of the site where we are bitten by Ixodes)
NUMBER • Contains linear chromosome.
Buffalo Bison bison 60 • Discovered by Hinnebusch and Tilly (1993)
Cat Felis catus 38
Cattle Bos taurus 60 CELLULAR DIVISION
Bos indicus
Dog Canis familiaris 78
Donkey Equus asinus 62
Goat Capra hircus 60
Horse Equus caballus 64
Human Homo sapien 46
Pig Sus scrofa 37
Sheep Ovis aries 54

POLYPLOIDY
• Having multiple sets of chromosomes.
• From the term itself “poly” means many, multiple sets of
chromosomes.
• The basics terminologies of chromosomes: We have what
we called; GAP 0 OR G0 PHASE
o Haploid which is a one (1) set of chromosomes • The resting or quiescent phase.
o Diploid which is a two (2) sets of chromosomes
• Inactive or non-cycling phase
o Triploid which is a three (3) of chromosomes
o Tetraploid four (4) sets of chromosomes
o Pentaploid which is a five (5) sets of chromosomes. INTERPHASE
• The non-dividing stage of the cell.
HOMOLOGOUS CHROMOSOMES • The longest stage in eukaryote cell division.
• In this phase replication of DNA occurs.

3 STAGES OF CELL DIVISION

Table No. 2 Three Stages of Cell Division
Subphase Processes
Gap 1 (G1) This is the Growth phase and
synthesis of amino acids
happens here (A.A) and other
biochemical needed for S
phase.
S phase (Synthesis) The DNA in chromosomes are
• Basic parts of chromosomes: replicated. Also, during this
o Which part of the chromosomes are the longest arm - phase, the cells are
Q arm synthesized.
o Which part of the chromosomes are the shortest arm? Gap 2 (G2) This is the production of
▪ P arm microtubules. Also, this is the
o The tip of chromosomes is called Telomere stages where the mitosis takes
o The center of the chromosomes is called Centromere place.
• We have what we called sister chromatid and non-sister
chromatid. To differentiate, the sister chromatid has 2 M PHASE (MITOSIS PHASE)
homologous chromosomes are joined together and non- • The dividing stage.
sister chromatid two different chromosomes.
2 STAGES
• Karyokinesis – division of the nucleus (1st step in M
phase).
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 TRANS: INTRODUCTION TO CYTOLOGY

• Cytokinesis – Division of the cytoplasm to form two

separate daughter cells immediately after mitosis (2nd
stage of M phase)

MITOSIS
• Nuclear division.
• (2) types:
o Somatic cells (blood cells, epithelial cells, bone cells,
and other differentiated cells).
o Reproductive cells (sperm cells and egg cells/ovum)
▪ Predominant sugar in sperm cell is Fructose
(fructose = fruits).

STAGES OF MITOSIS
PROPAHSE
• The nuclear membrane begins to disintegrate.
• Nucleoli disappear.
• DNA begin to supercoil and appear as chromosomes.
o Chromosomes move toward to the center.

METAPHASE
• Chromosomes appear as sister chromatids attached
together at the centromere, containing the kinetochore.
• Kinetochore are microtubules that bind chromosomes
(word means move or spread) these are located inside
centromere.
• Each chromosomes have 2 kinetochores.
• Centrioles appear on the both poles of the cells.
• Spindle fibers appear and attached to the kinetochores.
o Chromosomes align at the center of the cell.
o Kinetochore- found inside the centromere, 2
kinetochore per chromosomes
▪ Kineto means move
▪ Chore means spread
ANAPHASE
• Sister chromatids are pulled towards opposite poles.
• When sister chromatids are pulled together it will form a
cleavage.
TELOPHASE
• The nuclear membrane reappears.
• The DNA begin to diffuse into the nucleus.
• Nucleoli reappears.
o In here two daughter cells will appear.
o The chromatin fibers are uncoiled.
o The mitosis ends in this phase.
MEIOSIS
• A course of double division producing (4) daughter cells
with half the number of chromosomes as that of the parent
cells.
• This chromosome number is referred as “haploid” in
contrast to the parent cell which is “diploid.”
• Necessary for the sexual reproduction among eukaryotes.
• Has 9 stages which are:
1. Interphase
2. Prophase 1
3. Metaphase 1 (line up in the equator)
4. Anaphase 1 (pulled-apart)
5. Telophase 1 and cytokinesis (cell pinches in the
middle)
6. Prophase 2 (2 daughter cells)
7. Metaphase 2
8. Anaphase 2
9. Telophase 2 and cytokinesis (2 daughter cells)
• Same process with mitosis but the process in meiosis is x2
and they utilize the two daughter cells that came from
mitosis.
CHIASMATA
• A point of overlap of paired chromatids at which fusion and
exchange of genetic material takes place during prophase
of meiosis.
o When two overlaps each other they will be able to
exchange the genetic material from one another.

SUBSTAGES OF PROPHASE
• Leptotene
o The first stage of the prophase of meiosis, during which
each chromosome becomes visible as two fine threads
(chromatids) within the nucleus.
• Zygotene
o Homologous chromosomes pair up during synapsis.

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 TRANS: INTRODUCTION TO CYTOLOGY

▪ Remember that synapsis begins with

synaptonemal complex between two homologous
chromosomes
• Pachytene
o Crossing over happens.
o The paired chromosomes shorten and thicken, the two
chromatids of each separate, and exchange of
segments between chromatids may occur.
▪ Cross over of genetic material between two
chromatids happens in here.
▪ During this process we will have chiasmata
because you are able to exchange genetic
material.
• Diplotene
o Chromosomes separate slightly.
o The paired chromosomes begin to separate into two
pairs of chromatids.
• Diakinesis
o Nucleolus disappears.
o Nuclear envelope disintegrates completely.
o In this stage spindle fibers begin to form.

SPINDLE FIBERS
• They are chiefly involved in moving and segregating the
chromosomes during nuclear division.
• Spindle fibers are made up of microtubules.
o This is a protein structure that divides the genetic
materials since they are able to pull apart the cell from
both poles.

IMPORTANT NOTES
• Meiosis 1- reductional phase
• Meiosis 2- equational phase

REFERENCES

Notes from the discussion by: Sir. Roel R. Guinto Jr., RMT,
MSMT

National University powerpoint presentation:

C.J. ANIS, R. ARANGCO, C.D. GECALE, M.D.G. GOLEZ, A.J. MANGILIT | BSMT 5
 TRANS: INTRODUCTION TO CYTOLOGY

C.J. ANIS, R. ARANGCO, C.D. GECALE, M.D.G. GOLEZ, A.J. MANGILIT | BSMT 6