Exercise Number 3

CELL DIVISION

All living organisms consists of cells. Unicellular and multicellular organisms grow and
reproduce by cell division. The physical and metabolic activities of growing cells are cyclic – e. i.
they constitute a sequence of events that is dividing and repetitive as long as the cell is growing
and dividing. The cell cycle comprises essentially two periods: the interphase which consists of
G1, S and G2 phases; and the period of division which can either be by mitosis or meiosis.

INTERPHASE

The non-dividing cell has distinct nucleus enclosed by a definite nuclear membrane. Within
the nucleus are one or more small, round dense bodies called nucleoli and extremely long and thin
chromosomes which appear as a granular network of darkly staining material called chromatin.

This. On mitotic stage is divided into three distinct phases which are biochemically but not
morphologically distinguishable.

1. G1-phase – the first growth stage in which the nucleus and cytoplasm are enlarging toward
mature size.

2. S-phase the stage where the synthesis of DNA and histone occur. The DNA doubles in amount.
This is reflected in the chromosomes becoming longitudinally double so that each consists of two
chromatids.
3. G2 phase – the time of active synthesis of RNA and proteins necessary for chromosome
synthesis and for the mitotic spindle.

DIVISION

A. Mitosis

Sexually reproducing organisms usually possess two sets (2n) of chromosomes, one set (n)
received from the father and the other (n) from the mother. Normally, each of the chromosomes or
homologue in the other. Transmission of the chromosomes from the parent to offspring is
accomplished through fertilization of gametes. Gametes have only one set (n) of chromosomes.
The fertilized egg or zygote (2n) develops into a multicellular organism in which each cell typically
has the same number of chromosomes as the original zygotes. This is so because the cells have
divided. This chromosome constancy is accomplished by mitosis, an orderly continuous series of
events wherein each cell divides to form two cells which are exact replicas of the cell from which
they were derived.
Mitosis also occurs in individuals that are asexually reproducing and whose chromosome
numbers are not diploid. This process, therefore, is undergone by all somatic (body cells)
irrespective of their type of reproduction and chromosome number.
In multicellular organisms, mitosis is a means of increasing the number of cells and
replacing worn out tissues; in unicellular organisms, it is simply a mode of reproduction. Mitosis
can be divided into four distinct stages:
1. Prophase
The chromosomes shorten and thicken and become visible as thick rods. Each of the
chromosomes appear longitudinally split into two duplicates. Nucleoli and nuclear membrane
complete disappear by late prophase.

2. Metaphase
The chromosomes which have continued to get thicker and shorter line up on a single plane
midway between the poles of the spindle (metaphase plate). The spindle is composed in part of
protein fibers which play a role in chromosome movement during nuclear division. The duplicated
chromosomes are still attached to each other at the centromere which becomes associated with the
spindle fiber.
In animal cells, the spindle is usually formed between two centrioles that were formally
together on one side of the nucleus. As these centrioles separate and move to the new positions on
the opposite sides of the nucleus, they appear to radiate distinctive lines (astral rays). In many
plants, the centrosomes and their accompanying centrioles are missing although spindles are
nevertheless present.

3. Anaphase
The duplicated chromosomes separate at the centromere, forming their respective
centromeres which move apart. They toward opposite poles of the spindle where the arms of the
chromosomes are dragged along passively.

4. Telophase
Once the chromosomes are at the poles, the spindle disintegrates, a new nuclear membrane
is formed around the chromosomes and nucleoli reappear. The chromosomes begin to uncoil and
lengthen, and finally lose their visible identity.
Cytokinesis (cytoplasmic division) usually follows mitosis. In plant tissues, the cytoplasm
is divided via cell plate formation where separation starts from the inside of the cell towards the
periphery and in animal tissues via furrowing (this process works in opposite direction).
The most important feature of mitosis from the standpoint of the study of heredity is that
the chromosome number remains constant through successive cell divisions. There is a
chromosome duplication followed by cell division. The result is an exact distribution of
chromosomes to the new cells as they are formed, and the chromosome make up of a cell is the
same as that of the cell from which it has been divided.

B. Meiosis

When sexually reproducing individuals reach maturity, gametes (n) are formed in the
gonads of animals and in the sporangia of stamens and pistils of plants. Transmission of the
chromosomes from parent to offspring is accomplished through fertilization of gametes resulting
to a zygote (2n). Since a zygotic nucleus contains twice as many chromosomes as do the gametic
the gametic nuclei and since the chromosome number remains constant from generation to
generation, there must be a process which reduces the diploid chromosome complement back to
the haploid number. This is accomplished through meiosis which occurs during gametogenesis,
the whole process involved in gamete formation. In male animals, gametogenesis is specifically
termed spermatogenesis (sperm formation); in females, oogenesis (egg formation) in higher plants,
micro or mega sporogenesis (spore formation).
Meiosis is preceded by G1, S, and G2 phases of the cell cycle. It consists of two nuclear
divisions which follow each other in rapid sequence. The first involved the separation of
homologous chromosomes resulting in two haploid nuclei while the second results in four haploid
nuclei.

1. Meiosis I

a. Prophase I. The cells gradually increase in size. Just as mitotic prophase, each chromosome
contains two duplicates coiled about each other. There are several well defined substages.

(1) Leptonema. The chromosomes are long and thin, more so than in the earliest mitotic
prophase. Nucleolus is distinct.

(2) Zygonema. Corresponding (homologous) chromosomes pair with each other. The pairing
process, known as synapsis, is very exact, being not merely between homologous chromosomes
but between corresponding individual points. Synapsis proceeds zipperwise until the two
homologous are completely paired.

(3) Pachynema. The pairing threads are so intimately associated that it is difficult to see that
they are composed of two chromosomes. A synaptonemal complex forms between homologues.
This is an avenue of genetic crossing over (an actual exchange of corresponding segments between
homologous chromosomes).

(4) Diplonema. The tight pairing is relaxed, whereupon it can be clearly seen that each pair of
synapsed chromosomes contains four threads because each chromosome has itself doubled. A pair
of synapsed chromosomes is known as bivalent (composed of two univalent) when referring to
chromosomes.

Separation of paired chromosomes is still incomplete since they appear attached to each
other at various points along their length, each point is called a chiasma (plural chiasmata). The
chiasma is important since it is the physical basis for the genetic phenomenon of crossing over.
Chiasma terminalization (movement of chiasmata toward the end of chromosome arms i.e.
away from the centromere) takes place. The chromosomes also become thicker and shorter.
(5) Diakinesis. Bivalents attain maximal contraction and are distributed throughout the nucleus.
The chromosome number of the species could be counted. By the end of this stage, nucleoli and
nuclear membrane have disappeared, the spindle has formed, and Prophase I is completed.

b. Metaphase I. Bivalents align themselves at the equatorial plate.

c. Anaphase I. The univalent in each bivalent separate from each other at the region of the
centromere and proceed to opposite poles of the spindle.

d. Telophase I. Two haploid nuclei are formed. Nucleolus and nuclear membrane reappear;
chromosomes uncoil and lengthen. Cytokinesis may not occur in some species.

2. Meiosis II

a. Prophase II. Univalents contract, nucleolus and nuclear membrane disappear.

b. Metaphase II. Univalents line up at the equatorial plate.

c. Anaphase II. The members of a univalent separate and proceed to the poles as a s ingle
chromosomes.

d. Telophase II. Chromosomes uncoil and lengthen; nuclear membrane and nucleolus reappear.
Four haploid daughter nuclei are formed with the division of the cytoplasm.

Mitosis and meiosis are the mechanisms that permit the orderly distribution of
chromosomes to daughter cells and the orderly reduction in chromosome number, respectively.

OBJECTIVES

1. To differentiate the stages that an actively growing cell undergoes with emphasis on the
chromosomal changes.
2. To be able to determine the consequences of then different types of cell divisions in terms of
heredity.

MATERIALS

Prepared slides for mitosis – Allium cepa root tip and whitefish blastula
Prepared slides for meiosis – Rhomaleum testis

METHODS

I. Preparation of slide showing cells of onion in active mitosis.

1. With the help of a sharp blade, carefully snip the dry roots of the onion. Place the bulbs in a
beaker containing water to grow the root tips. It may take around 4 to 6 days for the new roots to
grow and appear

2. Trim around 3 cm of the newly grown roots and place them in a watch glass. With the help of
forceps, shift it to a vial holding freshly prepared aceto-alcohol i.e., a mixture of glacial acetic acid
and ethanol in the ratio 1:3. Allow the root tips to remain in the vial for one complete day.
3. Place the fixed root on a slide. Note the whitish tip of the root; this is the root cap. Cut the
terminal 1 mm of the root using a piece of razor to remove the root cap. Discard the root cap.
Actively dividing cells will be found in the meristematic region of the root, which follows the root
cp.

4. Cut a very thin section of the meristematic region. Discard the rest of the root.

5. Add one drop of acetocarmine on the thin root section. Let the preparation stand for 1 minute.

6. Cover the stained root section with coverslip and heat gently by passing it through the flame of
alcohol lamp. Heating facilitates staining of the chromosomes and clears the cytoplasm. Be careful
not to allow the specimen to dry out.

7. Gently tap the coverslip with the pointed end of a straight needle or with unused eraser end of a
pencil to spread the cells.

8. Observe the specimen under low power objective (LPO) to access if the chromosomes have
been properly stained.

a. If chromosomes are deeply stained, destain them using 45% acetic acid. Place a drop of acetic
acid on one side of coverslip and place a piece of filter paper against the opposite side of the
coverslip to draw the acetic acid across specimen. Heat the slide by passing it 3-4 times through
the flame to facilitate staining. Reassess the staining of the chromosomes after staining.

b. If the chromosomes are poorly stained, restain them using acetocarmine. Place a drop of
acetocarmine on one side of the coverslip and place a piece of filter paper against the opposite side
of the coverslip to draw the stain across the specimen. Heat the slide by passing 3-4 times through
the flame to facilitate staining. Reassess the staining of chromosomes after heating.

9. Once the chromosomes have been properly stained, seal the specimen with paraffin. Heat a bent
needle over the flame, place it on the paraffin and apply the melted paraffin to the edges of the
coverslip.

10. Using low power objective (LPO), look for cells undergoing mitosis. Switch to the high power
objective (HPO) to identify the stages of mitosis exhibited by the dividing cell. Observe the
arrangement and morphology of the chromosomes at each stage of mitosis. Draw one cell per stage
of mitosis.

Wire Models of Cell Division (Group Activity)

1, Get two long wires (one green and one red) and two short wire (one green and one red). These
would represent the original chromatin fibers in a cell. Wires of the same length are homologous
to each other. Red wires represent chromatin from the female parent of the cell and green wires
represent chromatin from its male parent.
2. Using tape, mark the centromere of each chromatin fiber; place the tape exactly at the middle
of the short wires, and at a point ¾ from one end of the long wires.

3. Using tape, label the chromatin at their tips as follows:

long green = 1 p long red = 1 m
short green = 2 p short red = 2m

4. Simulate what happens as the cell goes through the S phase. Replicate each chromatin fiber by
getting an identical set of wires. Connect sister chromatids to each other by placing tape around
their centromeres.

5. Convert the chromatin fibers into chromosomes by coiling the wires.

III. Microscopic examination of prepared slides of actively dividing cells.

1. Prepared slides of cells in active mitosis from the root tip of Allium cepa and whitefish blastula.
Identify all stages of mitosis as seen in your slides. Consult the instructor for the accuracy of
identification. Draw the different stages of mitosis both from plant and animal specimen.

2. Prepared slide observation for meiosis Rhomaleum testis

3. Identify all stages of meiosis seen in your slide. Consult your instructor for accuracy of
identification. Draw the various stages as seen under the microscope. Label all stages observed.
In a tabular form (Table 3.1), list down the difference between mitosis and meiosis.
 Exercise No. 3
CELL DIVISION

Name: ___________________ Score: ______

A. Stages of Mitosis

Interphase Prophase Metaphase

Anaphase Telophase Two Daughter Cells

Table 3.1 Differences between mitosis and meiosis

Criteria Mitosis Meiosis

Chromosome number of daughter cells
Number of cell division
Stages
Presence of synapsis
Presence of crossing over
Cell type that undergoes division
Number of daughter cells formed
DNA content of cells at the start of
division
DNA content of daughter cells
Genetic consequences

Questions:

1. In what aspect does mitotic prophase nucleus different from a daughter telophase nucleus?
2. What is the significance of mitosis? meiosis?
3. Why is meiotic division a must in gamete formation? What is its relationship to fertilization?
4. What is the relationship of meiosis with Mendelian Laws of Inheritance?
5. Define each of the following terms:
a. centromere
b. tetrad
c. genome
d. chromatin
e. cytokinesis
f. chromosome
g. chromatid
h. centriole
i. homologue
j. kinetochore
k. synapsis
l. crossing over
m. cytokinesis
n. autosomes
o. sex chromosome
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