The mitotic cell cycle

Nabeela Ashraff
Structure of chromosome
The importance of mitosis

• Mitosis is the process of nuclear division by which two genetically identical

daughter nuclei are produced that are also genetically identical to the parent
nucleus
• The process of mitosis is of great biological significance and is fundamental to
many biological processes:
• Growth of multicellular organisms
• The two daughter cells produced are genetically identical to one another
(clones) and have the same number of chromosomes as the parent cell
• This enables unicellular zygotes (as the zygote divides by mitosis) to grow
into multicellular organisms
• Growth may occur across the whole body of the organism or be confined to
certain regions, such as in the meristems (growing points) of plants
• Replacement of cells & repair of tissues
• Damaged tissues can be repaired by mitosis followed by cell division
• As cells are constantly dying they need to be continually replaced by
genetically identical cells
• In humans, for example, cell replacement occurs particularly rapidly in the skin
and the lining of the gut
• Some animals can regenerate body parts, for example, zebrafish can regenerate
fins and axolotls regenerate legs and their tail amongst other parts
• Asexual reproduction
• Asexual reproduction is the production of new individuals of a species by a
single parent organism – the offspring are genetically identical to the parent
• For unicellular organisms such as Amoeba, cell division results in the
reproduction of a genetically identical offspring
• For multicellular organisms (as seen with many plant species) new individuals
grow from the parent organism (by cell division) and then detach (‘bud off’)
from the parent in different ways. Some examples of these are budding
in Hydra and yeast and runners from strawberries
The cell cycle

• Mitosis is part of a precisely controlled process known as the cell cycle

• The cell cycle is the regulated sequence of events that occurs between one cell
division and the next
• The cell cycle has three phases:
• interphase
• nuclear division (mitosis)
• cell division (cytokinesis)

• The length of the cell cycle is very variable depending on environmental conditions, the
cell type and the organism
• For example, onion root tip cells divide once every 20 hours (roughly) but human intestine
epithelial cells divide once every 10 hours (roughly)

• The movement from one phase to another is triggered by chemical signals called cyclins
Interphase

During Interphase the cell increases in mass and size and carries out its normal cellular
functions (eg. synthesising proteins and replicating its DNA ready for mitosis)
• Interphase consists of three phases:
• G1 phase
• S phase
• G2 phase

• It is at some point during the G1 phase a signal is received telling the cell to divide again

• The DNA in the nucleus replicates (resulting in each chromosome consisting of two
identical sister chromatids)
• This phase of the interphase stage of the cell cycle is called the S phase – S stands
for synthesis (of DNA)
• The S phase is relatively short
• The gap between the previous cell division and the S phase is called
the G1 phase – G stands for gap
• Cells make the RNA, enzymes and other proteins required for
growth during the G1 phase

• Between the S phase and next cell division event the G2 phase occurs
• During the G2 phase, the cell continues to grow and the new DNA that
has been synthesised is checked and any errors are usually repaired
• Other preparations for cell division are made (eg. production of tubulin
protein, which is used to make microtubules for the mitotic spindle)
• Interphase = G1 + S + G2
 Nuclear division
• Follows interphase
• Referred to as the M
phase – M stands for mitosis
• Cell growth stops during the M
phase

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 Cytokinesis
• Follows M phase
• Once the nucleus has divided into two
genetically identical nuclei, the whole cell
divides and one nucleus moves into each
cell to create two genetically identical
daughter cells
• In animal cells, cytokinesis
involves constriction of the
cytoplasm between the two nuclei and in
plant cells a new cell wall is formed

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The stages of mitosis

• Mitosis is the process of nuclear division by which two genetically identical daughter
nuclei are produced that are also genetically identical to the parent cell nucleus (they have the
same number of chromosomes as the parent cell)
• Although mitosis is, in reality, one continuous process, it can be divided into four main stages
• These stages are:
• Prophase
• Metaphase
• Anaphase
• Telophase

• Most organisms contain many chromosomes in the nuclei of their cells (eg. humans have 46) but
the diagrams below show mitosis of an animal cell with only four chromosomes, for simplicity
• The different colours of the chromosomes are just to show that half are from the female parent
and half from the male parent
Exam tip

• Make sure you learn the four stages of mitosis. Cytokinesis is often mistaken as a stage of mitosis,
but remember this is a separate part of the cell cycle.
• The acronym PMAT can be helpful to remind you what happens during each stage of mitosis:
• P = Prophase, where the cell Prepares to divides
• M = Metaphase, where the chromosomes align along the Middle
• A = Anaphase, where the chromosomes move Away from each other
• T = Telophase, where Two nuclei reform
• The chromosome number is important too; after interphase but before the parent cell undergoes
mitosis, the human parent cell nucleus actually contains 92 DNA molecules! This is because during
interphase (S phase), the 46 DNA molecules in the parent cell have replicated to form sister
chromatids. As human cells have a diploid number of 46 this replication results in 92 molecules. This
ensures the two daughter cells will be diploid (have 46 chromosomes each) when mitosis occurs.
Remember to read the questions carefully as only human diploid cells have 46 chromosomes so if the
question refers to another organism, its diploid number will be different.
Prophase

• Chromosomes condense and are now visible when stained

• The chromosomes consist of two identical chromatids called sister
chromatids (each containing one DNA molecule) that are joined together at
the centromere
• The two centrosomes (replicated in the G2 phase just before prophase) move
towards opposite poles (opposite ends of the nucleus)
• Spindle fibres (protein microtubules) begin to emerge from
the centrosomes (consists of two centrioles in animal cells)
• The nuclear envelope (nuclear membrane) breaks down into small vesicles
Metaphase

• Centrosomes reach opposite poles

• Spindle fibres (protein microtubules) continue to extend from centrosomes
• Chromosomes line up at the equator of the spindle (also known as the
metaphase plate) so they are equidistant to the two centrosome poles
• Spindle fibres (protein microtubules) reach the chromosomes and attach to
the centromeres
• Each sister chromatid is attached to a spindle fibre originating from opposite
poles
Anaphase

• The sister chromatids separate at the centromere (the centromere divides

in two)
• Spindle fibres (protein microtubules) begin to shorten
• The separated sister chromatids (now called chromosomes) are pulled to
opposite poles by the spindle fibres (protein microtubules)
Telophase

• Chromosomes arrive at opposite poles and begin to decondense

• Nuclear envelopes (nuclear membranes) begin to reform around each set of
chromosomes
• The spindle fibres break down
Practical : Observing the stages of
mitosis
• Growth in plants occurs in specific regions called meristems
• The root tip meristem can be used to study mitosis
• The root tip meristem can be found just behind the protective root cap
• In the root tip meristem, there is a zone of cell division that contains cells
undergoing mitosis
• Pre-prepared slides of root tips can be studied or temporary slides can be
prepared using the squash technique (root tips are stained and then gently
squashed, spreading the cells out into a thin sheet and allowing individual
cells undergoing mitosis to be clearly seen)
Apparatus

• Roots of onion or garlic

• Microscope slide
• Cover slip
• 1 M hydrochloric acid
• Stain (acetic orcein, toluidine blue for example)
• Paper towel
• Scalpel blade
• Cutting tile
• Water bath at 60oC
• Boiling tube
• Optical microscope
Method

1.Garlic or onion (Allium cepa) root tips are most commonly used (the bulbs can
be encouraged to grow roots by suspending them over water for a week or two)
2.Prepare a boiling tube of 1M hydrochloric acid and place in a water bath at 60oC
for 10 minutes
3.Remove the tips of the roots (about 1cm) and place in the warmed
hydrochloric acid for 5 minutes
4.Rinse the tips well in cold water using a pipette and blot dry with a paper towel
5.Cut approximately 2mm off the tip and place onto a microscope slide
6. Add a drop of a suitable stain (eg. warm, acetic orcein, which stains
chromosomes a deep purple)
7. The stained root tip is gently squashed on a glass slide using a blunt
instrument (eg. the handle of a mounting needle or a cover slip using your fist to
gently apply pressure in a rolling motion)
8. View the slide under the microscope
9. Cells undergoing mitosis (similar to those in the images below) can be seen and
drawn
10. Annotations can then be added to these drawings to show the different
stages of mitosis
Results
Calculating Mitotic index

• The mitotic index is the proportion of cells (in a group of cells or a sample of
tissue) that are undergoing mitosis
• The mitotic index can be calculated using the formula below:
• mitotic index = number of cells with visible chromosomes ÷ total
number of cells
• You can multiply the answer by 100 if you need to give the mitotic index as
a percentage
Question

A student who wanted to observe mitosis prepared a sample of cells. They

counted a total of 42 cells in their sample, 32 of which had visible
chromosomes. Calculate the mitotic index for this sample of cells (give your
answer to 2 decimal places).

• Mitotic index = number of cells with visible chromosomes ÷ total number of

cells
• Mitotic index = 32 ÷ 42
• Mitotic index = 0.76
Question

The table below shows the number of cells in different stages of mitosis in a
sample from a garlic root tip. Calculate the mitotic index for this tissue (give your
answer to 2 decimal places).

• Mitotic index = number of cells with

visible chromosomes ÷ total number of
cells
• Mitotic index = (prophase + metaphase
+ anaphase + telophase) ÷ total
number of cells
• Mitotic index = (14 + 5 + 3 + 6) ÷ (36
+ 14 + 5 + 3 + 6)
• Mitotic index = 28 ÷ 64
• Mitotic index = 0.44
Question

The micrograph below shows a sample of cells from an onion root tip. Calculate
the mitotic index for this tissue (give your answer to 2 decimal places).
Answer
• Number of cells with visible chromosomes (green) = 20
• Total number of cells (green + red) = 20 + 55 = 75
• Mitotic index = number of cells with visible chromosomes ÷ total number of cells
• Mitotic index = 20 ÷ 75
• Mitotic index = 0.27
 The Significance of Telomeres
• The ends of the chromatids in chromosomes are ‘sealed’ with protective structures called telomeres
• They are made of non-coding DNA (DNA that does not contain genes) that is made up of short base
sequences that are repeated many times (multiple repeat sequences)
• In telomeres, one strand is rich in the base guanine (G) and the other strand is rich in the complementary base
cytosine (C)
• The main function of telomeres is to ensure that the very ends of the DNA molecules are included in DNA
replication during mitosis (the copying enzyme responsible for DNA replication is unable to run right to the very
end of the DNA molecule and stops a little short of the end)
• If this end part of the DNA molecule contained an important gene, that piece of genetic information would be
lost during DNA replication
• In each subsequent cell division, a little more genetic information would be lost
• Telomeres therefore act as a ‘buffer’ region of non-essential DNA and ensure that no important coding
sections near the ends of the DNA molecules are left out of the replication process
• This ensures no genes are lost during cell division (the loss of vital genes can even result in cell death)
and allows for continued replication of a cell
• To avoid the risk of losing genes most cells have an enzyme called telomerase that adds additional bases at
each end (the telomeres)
• Some cells (generally specialized cells) do not have telomerase to ‘top up’ their telomeres and therefore after a
certain number of cell divisions the cell dies, this has been connected with the ageing process
The significance of stem cells

• A stem cell is a cell that can divide (by mitosis) an unlimited number of
times
• Each new cell (produced when a stem cell divides) has the potential to remain
a stem cell or to develop into a specialised cell such as a blood cell or a
muscle cell (by a process known as differentiation)
• This ability of stem cells to differentiate into more specialised cell types is
known as potency
• There are three types of potency:
• Totipotency – totipotent stem cells are stem cells that can differentiate into any cell
type found in an embryo, as well as extra-embryonic cells (the cells that make
up the placenta). The zygote formed when a sperm cell fertilises an egg cell is
totipotent, as are the embryonic cells up to the 16-cell stage of human embryo
development
• Pluripotency – pluripotent stem cells are embryonic stem cells that can differentiate
into any cell type found in an embryo but are not able to differentiate into
extra-embryonic cells (the cells that make up the placenta)
• Multipotency – multipotent stem cells are adult stem cells that have lost some of
the potency associated with embryonic stem cells and are no longer pluripotent
Multipotent adult stem cells
• As tissues, organs and organ systems develop, cells become more and more specialized
• Having differentiated and specialized to fulfil particular roles, most adult cells gradually
lose the ability to divide until, eventually, they are no longer able to divide
• However, small numbers of stem cells (known as adult stem cells) remain to produce
new cells for the essential processes of growth, cell replacement and tissue repair
• Although these adult stem cells can divide (by mitosis) an unlimited number of times, they
are only able to produce a limited range of cell types – they are multipotent
• For example, the stem cells found in bone marrow are multipotent adult stem cells – they
can only differentiate into blood cells (red blood cells, monocytes, neutrophils and
lymphocytes)
• In adults, stem cells can be found throughout the body (eg. in the bone marrow, skin,
gut, heart and brain)
• Research is being carried out on stem cell therapy, which is the introduction of adult
stem cells into damaged tissue to treat diseases (eg. leukemia) and injuries (eg. skin
burns)
 How a Tumor Forms
• Cancers demonstrate how important it is that cell division is precisely controlled, as
cancers arise due to uncontrolled mitosis
• Cancerous cells divide repeatedly and uncontrollably, forming a tumour (an irregular
mass of cells)
• Cancers start when changes occur in the genes that control cell division. A change
in any gene is known as a mutation. If the mutated gene is one that causes cancer it is
referred to as an oncogene
• Mutations are common events and don’t lead to cancer most of the time
• Most mutations either result in early cell death or result in the cell being destroyed
by the body’s immune system
• As most cells can be easily replaced, these events usually have no harmful effect on
the body
• The mutations that result in the generation of cancerous cells do not result in early cell death or
in the cell being destroyed by the body’s immune system

• This means that the harmful mutation occurring in the original cell can be passed on to all that
cell’s descendants

• A typical tumor contains around a thousand million cancerous cells by the time it is detected

• Carcinogens are any agents that may cause cancer (eg. UV light, tar in tobacco smoke and X-rays).
If the agent causes cancer it is described as carcinogenic

• Some tumors (such as warts) do not spread from their original site – these are known
as benign tumors and do not cause cancer

• Some tumors spread through the body, invading and destroying other tissues – these are known
as malignant tumors and cause cancer
• Malignant tumors interfere with the normal functioning of the organ / tissue in which they have
started to grow (eg. they may block the intestines, lungs or blood vessels
• Malignant tumor cells can break off the tumor and travel through the blood and / or
lymphatic system to form secondary growths in other parts of the body
• The spreading of cancers in this way is known as metastasis
• Metastasis is very dangerous as it can be very difficult to detect, locate and
remove secondary cancers