Chapter – 10- Cell cycle and cell Division - Mitosis

INTRODUCTION:
 Growth and reproduction are characteristics of cells, indeed of all living organisms.

 All cells reproduce by dividing into two, with each parental cell giving rise to two daughter cells
each time they divide.

 These newly formed daughter cells can themselves grow and divide; giving rise to a new cell
population that is formed by the growth and division of a single parental cell and its progeny.
 In other words, such cycles of growth and division allow a single cell to form a structure
consisting of millions of cells.
HISTORY:
Name of the Scientists Contributions/role
1. Rudolf Virchow  Law of cell Lineage/Cell inheritance theory: “Omnis
cellula e cellula” New cells arise from pre-existing
cells.
2. Strasburger  First study of cell division in plants.
3. Walter Flemming  First study of cell division in animals.
4. Boveri and Flemming  Studied details of somatic cell division.
5. Flemming  Coined the term ‘Mitosis’.
6. Van Beneden  Discovered Meiosis
7. Sutton, Winiwater and  Studied details of Meiosis.
Strasburger
8. Farmer and Moore  Coined the term ‘Meiosis’.
9. Gregoire  Used term Meiosis I and Meiosis II.
TYPES OF CELL DIVISION:

i. Amitosis:
 It is characterized by the splitting of the nucleus followed by that of cytoplasm.
 It is seen in unicellular cell organisms like primitive algae, monerans, protozoans and the cells of
foetal membranes (membranes associated with the developing fetus).
 It does not involve the appearance of nuclear membrane, formation of chromosomes and
spindle and hence called direct cell division.
 **It starts with the elongation of the nucleus followed its division.
 **Amitosis does not involve nuclear events (prophase, metaphase, anaphase and telophase)
 **Spindle fibres are not involved.
 ii. Mitosis:
 The term mitosis was proposed by Flemming & its detailed study was given taken up A.
Schneider.
 Mitosis produced genetically identical cells, which are similar to mother or parental cell.
 **It is also called equational division or homeotypic division.

 It takes place when new cells are added to multicellular organisms as they grow and when tissues
are repaired or replaced.
 **Root tips (Root apical meristem) of onion (Allium cepa, 2n = 16) are best plant material for
the study of mitosis in labs.
 Root tips of Vicia faba (broad bean) are also used.
 In animals, cells at base of nail, bone marrow cells and skin cells (stratum germinativum) are
taken to study mitosis.
 **Acetocarmine is a nuclear basic stain used to study the cell division in plant material.
CELL CYCLE:
 The entire sequence of events which takes place in a cell between one cell division and the next.
 Cell division is a very important process in all living organisms.
 During the division of a cell, DNA replication and cell growth also take place.
 **All these processes, i.e., cell division, DNA replication, and cell growth, have to take place in a
coordinated way to ensure correct division and formation of progeny cells containing intact
genomes.
 The sequence of events by which a cell duplicates its genome, synthesizes the other constituents
of the cell and eventually divides into two daughter cells is termed cell cycle.
 Although cell growth (in terms of cytoplasmic increase) is a continuous process, DNA synthesis
occurs only during one specific stage in the cell cycle (S- phase).
 The replicated chromosomes (DNA) are then distributed to daughter nuclei by a complex series
of events during cell division.
 These events are themselves under genetic control. Complete life cycle of a cell is called cell cycle.
 PHASES OF CELL CYCLE:
 A typical eukaryotic cell cycle is illustrated by human cells in culture. These cells divide once in
approximately every 24 hours.
 Yeast (Saccharomyces cerevisiae – unicellular fungus) can progress through the cell cycle in only
about 90 minutes/1.30 minutes.
 **The time period of cell cycle is varied from organism to organism and also from cell type to
cell type.

Figure: The phases of the mitotic cell cycle.

 The correct sequence of a cell cycle is – (I Play Mostly AT Cricket ground)
 (Interphase) G1 →S →G2 →M→ Prophase →Metaphase→ Anaphase → Telophase →
Cytokinesis.
 Cell Cycle of an ordinary animal cell is –

STAGES OF CELL CYCLE

I. Interphase – Non-dividing, Non -resting phase: -
 This is phase between two successive M-phases.
 It is significant to note that in the 24-hour average duration of cell cycle of a human cell, cell
division (M-phase) proper lasts for only about an hour.

 The interphase, though called the resting phase, is the time during which the cell is preparing for
division by undergoing both cell growth and DNA replication in an orderly manner.
 In interphase cell grows in size and prepares itself for next division.
 **Interphase is most active phase of cell cycle misnomerly called resting phase.
 The interphase lasts more than 95% of the duration of cell cycle – 23 hours in human.
 Interphase is metabolically very active and important phase -DNA, RNA, Proteins, Fats,
Carbohydrates are synthesized, energy is stored and cell organelles doubled.
 Howard and Pelc classified interphase into 3 sub stages: -
(i) G1 – phase or Post-mitotic or Pre DNA synthesis phase (1st Gap phase) (Longest phase of cell
cycle)
 G1 phase corresponds to the interval between mitosis and initiation of DNA replication.
 During G1 phase the cell is metabolically active and continuously grows but does not replicate its
DNA.
 **During G1-most of cell organelles increases in cell and cell rapidly synthesizes different types
of RNA (RNA polymerase) (r- RNA; m- RNA; r- RNA) and proteins- synthesis of non-histone
proteins (Peptidyl transferase) - Heterocatalysis.
 Due to availability of protein, synthesis of new protoplasm takes place in cell and it starts growing
in size.
 **Cell grows maximum in G1 stage.

(ii) S – phase (DNA Synthesis phase or Replication phase- Autocatalysis):

 **It is the longest phase in interphase
 **Replication of nuclear DNA – DNA polymerase and synthesis of histone protein takes place in
S-phase.
 Replication of cytoplasmic or extranuclear DNA (It is now known that small circular
chromosomes, called extranuclear, or cytoplasmic, DNA, are located in two types of organelles
found in the cytoplasm of the cell. These organelles are the mitochondria in animal and plant
cells and the chloroplasts in plant cells.) may occur in the G1 phase of cell cycle.
 **During this time the amount of DNA per cell doubles. If the initial amount of DNA is denoted
as 2C (C-value is the amount, in picograms, of DNA contained within a haploid nucleus) then it
increases to 4C.
 However, there is **no increase in the chromosome number; if the cell had diploid or 2n number
of chromosomes at G1, even after S phase the number of chromosomes remains the same, i.e., 2n.
 S-phase marks the phase of DNA replication and chromosome duplication (DNA content in a
chromosome become doubled).
 **In animal cells, during the S phase, DNA replication begins in the nucleus, and the centriole
duplicates in the cytoplasm.
 Centriole replicates in late S-phase.
  Amount of DNA: G1 (2C = 50%) →S (4C) → G2 (4C) → Prophase (4C) →Metaphase
(4C) → Anaphase(2C) → Telophase(2C) → 2 daughter cells - 2C in each cell.
 Percentage of DNA transferred to daughter cells from S – phase of a mitocyte is
- 50 %
 In G1 phase of interphase, the number of chromosomes and amount of DNA in human
cell is 46 and 2C respectively. In G2 phase the number of chromosomes and amount of
DNA respectively is - 46 and 4C
(iii) G2 – phase (2nd Gap phase) or Post DNA synthesis phase (Pre mitosis phase)
 **Actual preparation (Final preparation) of M-phase occurs during this phase.
 Special materials required for M-phase are synthesized in G2 phase. eg. Tubulin protein. –
Required for formation of spindle fibres.
 During this phase proteins are synthesized in preparation for mitosis while cell growth continues.
 After G2 phase cell enters in division or M–phase.

(iv) G0 – phase (Quiescent stage):

 Some cells in the adult animals do not appear to exhibit division (e.g., heart cells) and many
other cells divide only occasionally, as needed to replace cells that have been lost because of injury
or cell death.
 These cells that do not divide further exit G1 phase to enter an inactive stage called quiescent
stage (G0) of the cell cycle.
 **Cells in G0 stage remain metabolically active but no longer proliferate unless called on to do so
depending on the requirement of the organism.

 Protein synthesis occurs in all 3 phases of interphase but DNA replication occurs
only in S- phases.
 In animal cells, during the S phase, DNA replication begins in the nucleus, and the
centriole duplicates in the cytoplasm.
 Tumour cells behave differently than normal cells in the body.
 They grow and divide in an uncontrolled manner (actively proliferating) and fail to respond to
signal.
 However, there are cells that become inactive and reside in quiescent phase (G0).
 These cells are known as quiescence cells that are less sensitive to drug treatments (radiotherapy
and chemotherapy) than actively proliferation cells.

 **Some cells like meristematic cells in plants and epidermal cells in the skin of mammals
undergo cell cycles continuously.
 Some cells like nerve cells, muscle cells, RBC are withdrawing from G1 phase and enter
permanently; into non dividing phase called G0 phase.
 Some cells like immuno-competent B-cells, T-cells etc., enter into temporary G0 phase and when
activated they reenter in to cell cycle.

II. M-phase (Mitotic) – Dividing phase :-( Shortest phase in cell cycle)
 **Division phase or M–phase or mitotic phase lasts for only about an hour-5% in the 24hour
duration of cell cycle of a human cell.
 The M-phase represents the phase when the actual cell division or mitosis occurs.
 **This is the most dramatic/distinct period of the cell cycle, involving a major reorganization of
virtually all components of the cell.
 Since the number of chromosomes in the parent and progeny cells is the same, it is also called
equational division.
 **The M-phase starts with nuclear division, corresponding to the separation of daughter
chromosome (Karyokinesis) and usually ends with division of cytoplasm (cytokinesis).
 **DNA replication, Karyokinesis, division of centromere/chromosome replication &
cytokinesis occurs only once.
 **The cell which undergoes mitotic cell division is called “Mitocyte”
 In animals, mitotic cell division is only seen in the diploid somatic cells (2n).
 Some social insects like male honey bees (Drones), haploid cells (n) divide by mitosis.
 In plants mitotic divisions occurs in both haploid and diploid cells.
 Though for convenience mitosis has been divided into four stages of nuclear division.

(1) PROPHASE: :(Longest stage in M-phase)

 **Prophase which is the first stage of karyokinesis of mitosis follows the S and G2 phases
of interphase.
 In the S and G2 phases the new DNA molecules formed are not distinct but intertwined.
 **Prophase is marked by the initiation of condensation/shrinkage of chromosomal
material.
 The chromosomal material becomes untangled during the process of chromatin condensation.
Chromatin threads condenses to form mitotic chromosomes
 Metabolism of cell decreases, cytoplasm becomes viscous, refractive and pale.

ANASTRAL AND AMPHIASTRAL MITOSIS:

 **In animals, the centriole, which had undergone duplication during S phase of
interphase, now begins to move towards opposite poles of the cell.
 **Astral ray (star shaped) forms due to gelation of proteins around centrioles.
 In animals, the asters are present and the mitosis is described as amphiastral, or astral
mitosis
 **In higher plants, centrioles are absent and no asters are formed. Mitosis without asters
is known as anastral mitosis.
 CHARACTERISTIC EVENTS OF PROPHASE:
i. At early prophase chromosomal material/chromatin fibres condenses to form compact
mitotic chromosomes.
ii. At mid prophase chromosome arms divide vertically except at centromere, hence each
chromosome has two chromatids attached together at the centromere.
iii. Initiation of the assembly of mitotic spindle, the microtubules, the proteinaceous
components of the cell cytoplasm help in the process.
iv. At late prophase when viewed under the microscope, do not show Golgi complexes,
endoplasmic reticulum nucleolus and nuclear envelope.
(2) METAPHASE:
 ** It is second stage of karyokinesis of mitosis follows the prophase.
 In this stage nuclear envelope/membrane undergoes complete
disintegration/degeneration hence the chromosomes are spread through the cytoplasm of
the cell.
 **Maximum condensation of chromosomes is completed and they can be observed clearly
under the microscope.
 **It is the best stage to study morphological structure of chromosomes.
 At this stage, metaphase chromosome is made up of two sister chromatids, which are held
together by the centromere.
 Small disc-shaped structures at the surface of the centromeres are called kinetochores.
 Kinetochores serve as the sites of attachment of spindle fibres - 97% tubulin protein and
3% RNA to the chromosomes that are moved into position at the centre of the cell.
 Metaphase is also characterised by all the chromosomes coming to lie at the equator with one
chromatid of each chromosome connected by its kinetochore to spindle fibres from one pole
and its sister chromatid connected by its kinetochore to spindle fibres from the opposite
pole.
 The plane of alignment of the chromosomes (centromeres) at metaphase is referred to as the
metaphase/equatorial plate.
 If 2n = 4 chromosomes, then
 Total number of chromosomes – 4
 Total number of centromeres – 4 (1 for each chromosome)
 Total number of chromosome arms – 8 (2 for each chromosome)
 Total number of kinetochores – 8 (2 for each chromosome)
 Total number of sister chromatids/DNA strands – 8 (2 for each chromosome)
 Total number of chromatid arms – 16 (4 for each chromosome)
CHARACTERISTIC EVENTS OF METAPHASE:
i. Nuclear envelope/membrane undergoes complete disintegration/degeneration &
chromosomes are released into cytoplasm.
ii. Maximum condensation of chromosomes is completed
iii. Spindle fibres attach to kinetochores of chromosomes.
iv. Formation of single metaphase/equatorial plate & single bipolar spindle apparatus.
v. Chromosomes are moved to spindle equator and get aligned along metaphase plate
through spindle fibres to both poles.ie., Centromere lies at equator and arms remain
directed towards poles
vi. Ca2+ and Mg2+ ions are necessary for assembly of microtubules.
(3) ANAPHASE: :(Shortest stage in M-phase)
 At the onset (the beginning) of anaphase, each chromosome arranged at the metaphase
plate is split simultaneously due to division of centromere and the two daughter
chromatids, now referred to as daughter chromosomes of the future daughter nuclei,
begin their migration towards the two opposite poles.
 As each chromosome moves away from the equatorial plate, the centromere of each
chromosome remains directed towards the pole and hence at the leading edge (the
foremost edge), with the arms of the chromosome trailing behind (the rear edge of a
moving body).
  **Approximately 30 ATP are required to carry a chromosome to pole. Chromosomes
reach at poles in late anaphase.
 Anaphase PromoComplex (APC) is a protein degradation machinery necessary for
proper mitosis of animal cells. If APC is defective in a human cell chromosomes will not
segregate.
If 2n = 4 chromosomes, then
 Total number of daughter chromosomes (=chromatids) inside the parental cell – 8
 Total number of daughter chromosomes (=chromatids) at each pole – 4
 Total number of centromeres – 4 (1/2 for each daughter chromosome)
 Total number of chromosome arms – 8 (2 for each daughter chromosome)
 Total number of kinetochores – 8 (1 for each daughter chromosome)
 Total number of sister chromatids – 8 (2 for each daughter chromosome)
 Number of DNA strands found at each pole – 8 (2 for each daughter chromosome)
 Total number of chromatid arms – 16 (2 for each daughter chromosome)

CHARACTERISTIC EVENTS OF ANAPHASE:

i. Centromeres split/divide and form 2 chromatids (=daughter chromosome) from each
parental chromosome.
ii. Chromatids move to opposite poles.
iii. The chromosomes at this stage may look like the shape of alphabets V (Metacentric),
L(Submetacentric), J(Acrocentric) and I(Telocentric) depending upon the position
of centromere in them.
(4) TELOPHASE (Reverse/opposite to prophase):
 At the beginning of the final stage of mitosis/karyokinesis, i.e., telophase, the
chromosomes that have reached their respective poles decondense and lose their
individuality.
 The individual chromosomes can no longer be seen and chromatin material tends to
collect in a mass in the two poles.

If 2n = 4 chromosomes, then
 Total number of chromatin fibres inside the parental cell – 8
 Total number of chromatin fibres at each daughter nucleus – 4
 Total number of daughters nuclei-2
 Total number of daughter nucleus at each pole-1
CHARACTERISTIC EVENTS OF TELOPHASE:
i. Chromosomes cluster at opposite spindle poles and their identity is lost as discrete
elements.
ii. Nuclear envelope assembles around the chromosome clusters.
iii. Nucleolus, Golgi complex and ER reform/reappears.
III. Cytokinesis (Division of cytoplasm):
 Mitosis accomplishes not only the segregation of duplicated chromosomes into daughter
nuclei (karyokinesis), but the parental cell itself is divided into two daughter cells by the
separation/division of cytoplasm called cytokinesis at the end of which cell division gets
completed.
a) Cytokinesis in animal cells by constriction/cleavage furrow method:
 In animal cells cytokinesis takes place by cell “furrow method.
 It is achieved by the presence of a furrow in the plasma membrane.
 The furrow gradually depends and ultimately joins in the centre dividing the cell
cytoplasm into two.
 In animals’ cytokinesis occurs in centripetal order i.e., from periphery towards centre.

b) Cytokinesis in plants by cell plate method:

 Cytokinesis in plants takes place by cell plate formation because constriction is not
possible due to presence of the rigid cell wall.
 Plant cells however, are enclosed by relatively inextensible cell wall, therefore they undergo
cytokinesis by a different mechanism.

 In plant cells, wall formation starts in the centre of the cell and grows outward to meet
the existing lateral walls of parental cell.
 In plants, cytokinesis occurs in centrifugal order (cell plate formation is from centre to
periphery).
 The formation of the new cell wall begins with the formation of a simple precursor, called
the cell-plate that represents the middle lamella between the walls of two adjacent cells.
 **Many Golgi vesicles and spindle microtubules arrange themselves on equator to form
phragmoplast – Barral shaped structure formed by microtubules at the centre of cell.
 Membrane of Golgi vesicles (Phragmosomes) fuse to form a plate like structure called
cell plate.
 **Golgi vesicles secret calcium and magnesium pectate. Further cell plate is modified into
middle lamella.
 **At the time of cytoplasmic division, organelles like mitochondria and plastids get
distributed between the two daughter cells.
 Multinucleate condition or syncytium:
 **In some organisms karyokinesis is not followed by cytokinesis as a result of which
multinucleate condition arises leading to the formation of syncytium
 Example: Liquid (coconut water) endosperm in coconut is a kind of free nuclear
endosperm contains thousands of nuclei.
 Coconut meat/kernel is a kind of cellular endosperm.ie., Dimorphic endosperm is found
in coconut
SIGNIFICANCE OF MITOSIS:
1. Mitosis or the equational division is usually restricted to the diploid cells only. However,
in some lower plants and in some social insects like male honey bees/drones (n=16)
haploid cells also divide by mitosis.
2. Mitosis usually results in the production of diploid daughter cells with identical genetic
complement.
3. The growth of multicellular organisms is due to mitosis.
4. Cell growth results in disturbing the ratio between the nucleus and the cytoplasm. It
therefore becomes essential for the cell to divide to restore the nucleo-cytoplasmic ratio.

5. A very significant contribution of mitosis is cell repair.

6. In animals’ cells of the upper layer of the epidermis, cells of the lining of the gut, and
blood cells are being constantly replaced.
7. Mitotic divisions in the meristematic tissues – the apical and the lateral cambium, result in
a continuous growth of plants throughout their life.
STAGES OF CELL IMPORTANT EVENTS
CYCLE
1. Interphase – Non-  Cell grows in size and prepares itself for next division.
dividing phase - Longest  Longest phase of cell cycle lasts more than 95% of the
phase in cell cycle. duration of cell cycle – 23 hours in human.
 Metabolically very active and important phase -DNA,
RNA, Proteins, Fats, Carbohydrates are synthesized,
energy is stored and cell organelles doubled.
a. G1- phase- Longest  Number of cell organelles increases in cell.
phase in interphase.  Cell rapidly synthesizes different types of RNA (RNA
polymerase) (r- RNA; m- RNA; r- RNA) and
proteins- synthesis of non-histone proteins (Peptidyl
transferase) - Heterocatalysis.
 Cell grows maximum.
b. S- phase – Longest  Replication nuclear (DNA content in a chromosome
synthetic phase in become doubled 2C to 4C).
interphase.  DNA - replication, synthesis of histones and formation
of new nucleosomes
 In animal cells, during the S phase, DNA replication
begins in the nucleus, and the centriole duplicates in
the cytoplasm.
c. G2- phase - Shortest  Actual preparation (Final preparation) of M-phase
phase in interphase. occurs during this phase.
  Tubulin protein. – Required for formation of spindle
fibres are synthesized in preparation for mitosis while
cell growth continues.
2. M- phase – Dividing  Most dramatic/distinct period of the cell cycle.
phase  Division phase or M–phase or mitotic phase lasts for
only about an hour-5% in the 24hour duration of cell
cycle of a human cell.
 It is the phase of shortest time in cell cycle.
I. Karyokinesis  Chromatin fibres condenses to form compact mitotic
a. Prophase - Longest chromosomes.
phase in M-phase.  Formation of two chromatids attached together at the
centromere.
 Initiation of the assembly of mitotic spindle
 Disappearence of Golgi complexes, endoplasmic
reticulum, nucleolus and nuclear envelope/membrane.
b. Metaphase  Nuclear envelope/membrane undergoes complete
disintegration/degeneration & chromosomes are
released into cytoplasm.
 Maximum condensation of chromosomes is completed.
 Best stage to study morphology of chromosomes
 Spindle fibres attach to kinetochores of chromosomes.
 Formation of single metaphase/equatorial plate &
single bipolar spindle apparatus.
 Centromere lies at equator and arms remain directed
towards poles
c. Anaphase - Shortest  Centromeres split/divide and form 2 chromatids
phase in M- phase (=daughter chromosome) from each parental
chromosome.
 Number of chromosomes doubled inside the cell.
 Chromatids move to opposite poles.
 The chromosomes at this stage may look like the shape
of alphabets V (Metacentric), L(Submetacentric),
J(Acrocentric) and I(Telocentric) depending upon the
position of centromere in them.
 Best stage to study various shapes of chromosomes
d. Telophase  Also called reverse prophase.
 Compact mitotic chromosomes undergoes
decondenses to form chromatin fibres.
 Chromosomes cluster at opposite spindle poles and
their identity is lost as discrete elements.
 Nuclear envelope Nucleolus, Golgi complex and ER
reform/reappears.
 Formation of two daughter nuclei inside the cell.
II. Cytokinesis  Formation of two daughter cells, each with same
number of chromosomes like that of parental cell.
 NUMERICALS AND SOLUTION IN MITOSIS:
1. Number of mitotic cell cycles/generations required to form 512 cells – 9

Solution: 2n – n- no of generations (=29)

 The question means how many times a single parent cell divides to give 512 cells.
 The total number of cells formed in a mitotic division = N X 2^n (We take 2 in the formula
because 1 cell divides to give 2 cells).
 where N is the number of cells undergoing mitosis and n is the number of cycles in a mitotic
division.
 Thus N = 1 (As 1 parent cell is undergoing mitosis).
 The total number of cells formed = 512.
 So, 1 X 2n = 512;2n = 512
 n = 9 cycles
2. After 6 generations of mitosis from a single cell, half of the cells are found entering G0
phase and the remaining half undergo 3 more generations of mitotic divisions. The
total number of cells of that system are – 288

3. Number of spindle apparatii formed during Rhizobium cell cycle – 0

Solution: Rhizobium is unicellular prokaryotic bacterium undergoes direct/amitotic cell
division.
4. Number of spindle apparatii formed during the formation of 512 cells – 511
Solution:Number of spindle apparatii formed during cell cycle = n-1; n= Total number cells-
1
5. Number of metaphasic plates found in metaphase of a Haplopappus mitocyte with
4 chromosomes – 1
Solution:Number of metaphasic plates during cell cycle = n-1; n= n= Total number cells-1