Cytogenetics

College of Allied Health Sciences

Bachelor of Science in Medical Laboratory Science
First Semester, A.Y. 2022-2023

[TRANS] THE CELL || THE CELL CYCLE

 Condenser
MICROSCOPY o Focuses light on the specimen and controls the light
 ZACCHARIAS JANSSEN for uniform illumination
o First inventor of the compound microscope  Light Source (Rheostat)
o Issue: poor quality microscope o Regulate the intensity of the light
 JOSEPH JACKSON LISTER
o Made a better compound microscope after Janssen BRIGHTFIELD
 Distinguishing Feature:
MAGNIFICATION o Uses visible light as source of illumination; cannot
 Ratio of an object's image to its real size resolve structures smaller than about 2 μm;
 Total Magnification specimen appears against a bright background.
o Computed by multiplying the magnification of the Inexpensive and easy to use
objective lens by the ocular lens  Principal Uses:
o Mostly used magnification of ocular lens: 10x o To observe various stained specimens and to count
MAGNIFICATION OBJECTIVES microbes; does not resolve very small specimens
Objective Magnification such as viruses
Scanner 4x
Low Power Obj 10x DARKFIELD
High Power Obj 40x  Distinguishing Feature:
Oil Immersion Obj 100x o Uses a special condenser with an opaque disk that
blocks light form entering the objective lens directly;
light reflected by specimen enters the objective lens
RESOLUTION / RESOLVING POWER and the specimen appears light against a black
background
 Measure the clarity of the image
 Principal Uses:
 It is the minimum distance between two points can be
o To examine living microorganisms that are invisible
separated and still be distinguished as separate points
in brightfield microscopy, do not stain easily, or are
 It is the ability of the lenses to distinguish fine detail
distorted by staining; frequently used to detect
structure
Treponema pallidum in the diagnosis of syphilis

CONTRAST
PHASE-CONTRAST
 The difference in brightness between the light and dark
 Distinguishing feature:
areas of an image
o Uses a special condenser containing an annular
 Has better contrast if we change the refractive index of the
(ringshaped) diaphragm. The diaphragm allows light
specimen
to pass through the condenser, focusing light on the
o Refractive Index- measure of the light bending
specimen and a diffraction plate in the objective lens.
ability of the medium
Direct and reflected or diffracted light rays are
o Staining Technique- technique used to change the
brought together to produce the image. No staining
refractive index
required.
o Forms halo around the image
PARTS  Principal Uses:
 Lens System o To facilitate detailed examination of the internal
o Ocular Lens – further magnification structures of living specimens
o Objectives – performs initial magnification o Useful in examining living unpigmented cells
o Course Adjustment Knob – moves mechanical stage

noticeably
DIFFERENTIAL INTERFERENCE MICROSCOPE (DIC)
o Fine Adjustment Knob – sharpens the image
 Distinguishing feature:
 Illumination System
o Like phase-contrast, uses differences in refractive
o Light Spurce
indexes to produce images. Uses two beams of light
o Condenser
separated by prisms; the specimen appears colored
o Iris Diaphragm
as a result of the prism effect. No staining required.
o Field Diaphragm
 Principal Uses:
 Body System
o To provide Three-dimensional images
o Base
o a.k.a Nomarski Microscopy/ Nomarski Interference
o Body Tube
Contrast
o Revolving Nose Piece
o Good in resolution compared to phase-contrast
 Note: Technique in Microscopy
o Can give almost or nearly threedimensional image
LPO → Coarse Adjustment Knob (isagad, hindi
o Interference Contrast
mababasag basta naka LPO) → slowly move downward
 Modulation Contrast Hoffman
until image becomes clear (tapos na, okay na)
 Differential Interference Contrast Nomarski
Additional: LPO → HPO (Parfocal/ Parfocal Distance)
 Brightfield Microscope can be a Modulation
 Contrast and Interference Contrast

ALVAREZ RMT’26 1
 Throughout Heaven and Earth, I alone am the Honored One

FLUORESCENCE  enabling today's researchers to know which cell fraction

 Distinguishing Feature: they should collect in order to isolate and study particular
o Uses an ultraviolet or nearultraviolet source of organelles.
illumination that causes fluorescent compounds TYPES OF MICROSCOPE
(green-colored) in a specimen to emit light MICROORGANISMS
 Principal Uses: Acellular Cellular
o For fluorescent-antibody techniques VIRUSES Prokaryotes Eukaryotes
(immunofluorescence) to rapidly detect and identify - Not Eubacteria

Cyanobacteria

Archaebacteria

Parasites

Fungi

Cell
Animal
microbes in tissues or clinical specimens cellular Gram Gram
 Fluorochromes- fluorescent compounds/fluorescent dyes but only
(+) (-)
consider
 Auramine O (used in Mycobacterium tuberculosis) ed as an Color Color
o Is a diarylmethane dye used as a fluorescent stain.

and
infectiou blue or red or
In its pure form, Auramine O appears as yellow s agent purple pink
needle crystals. It is insoluble in water and soluble in under under

Plant
ethanol and dimethyl sulfoxide micros micros
 Fluorecein Isothiocyanate (FITC)- used in Bacillus cope cope
anthracis (causative agent of anthrax)
 Anthrax- a serious infectious disease caused by gram- 3 MAJOR KINGDOMS
positive, rod-shaped bacteria known as Bacillus anthracis.  Bacteria
Anthrax can be found naturally in soil and commonly affects o Lack a membrane-bounded nucleus and
domestic and wild animals around the world. mitochondria, are surrounded by a cell wall, and
divide by binary fission
CONFOCAL o Example: most bacteria and cyanobacteria (blue-
 Distinguishing Feature: green algae)
o Uses a photon to illuminate one plane of a specimen  Archaea
at a time o Cell walls lack PEPTIDOGLYCAN
 Principal Uses o Share some common characteristics with bacteria.
o To obtain two-and three- dimensional images of the o Can stain gram (+) and gram (-)
cell for biomedical applications o Structure of the cell envelope and enzymes allows
them to survive under stressful or extreme conditions
ELECTRON MICROSCOPES  Eukarya
 Distinguishing Feature: o Cells contain an elaborate network of internal
o Uses a beam of electrons instead of light; electrons membranes, a membrane-bounded nucleus, and
pass through the specimen; because of the shorter mitochondria.
wavelength electrons, structures smaller than 2 μm o DNA is organized into true chromosomes, and cell
can be resolved. The image produced is two- division takes place by means of mitosis
dimensional. o Example: plants, animals, fungi and single-celled
 Principal Uses: organisms
o To examine viruses of the internal ultrastructure in
thin sections of cells (usually magnified 10,000-
100,000x)

SCANNING
 Distinguishing Feature:
o Uses a beam of electrons instead pf light; electrons
are reflected from the specimen; because of the
shorter wavelength of electrons, structures smaller
than 2 μm can be resolved. The image produced
appears threedimensional
 Principal Uses:
o To study the surface features of cells and viruses
(usually magnified 1000-10,000x)

CELL FRACTIONATION
 APPLICATION: Used to isolate or fractionate cell
components based on size and density
 TECHNIQUE: Cells are homogenized in a blender to break
them up. The resulting mixture is centrifuged. The
supernatant (liquid) is poured into another tube and
centrifuged at a higher speed for a longer period. This
process is repeated several times. This "differential
centrifugation" results in a series of pellets, each containing
different cell components.
 RESULTS: In early experiments, researchers used
microscopy to identify the organelles in each pellet and
biochemical methods to determine the metabolic functions.
These identifications established a baseline for this method,
ALVAREZ RMT’26 2
 Throughout Heaven and Earth, I alone am the Honored One

PROKARYOTES VS EUKARYOTES Gram (+) Gram (-)

PROKARYOTE EUKARYOTE Violet Primary Purple Purple
Nuclear Body NO NUCLEAR Classic Stain
MEMBRANE Membrane (Crystal
Cell Division Binary Fission Mitosis Violet)
Cell Wall (Eubacteria) (Animals, Iodine Mordant Purple Purple
Peptidoglycan Protozoans) (Iodine)
WITHOUT CELL Alcohol Decolorizer Purple Colorless
WALL (Alcohol)
(Archaebacteria) (Plants, fungi) Safranin Secondary Purple Red
Resembles WITH CELL Stain
Peptidoglycan WALL (Safranin)
Cytoplasmic (Phospholipid (Phospholipid
Membrane bilayer) without bilayer) with
CHO and Sterol CHO and Sterol
component component
Cell Organelles ABSENT Present
Site of Energy Cytoplasmic Mitochondria
Production Membrane
Site of Protein Free Ribosomes Rough ER
Synthesis

 BACTERIA – Single-Cell Prokaryotic Organisms

 FUNGI – Single-Cell or Multicellular Eukaryotic
Microorganisms
 YEASTS – Unicellular, Eukaryotic Microorganism
 PARASITES – Single-cell or Multicellular Eukaryotic
Microorganisms
 VIRUSES – Dependent on the host cells for survival and
therefore are NOT CONSIDERED CELLULAR
ORGANISMS but rather, INFECTIOUS AGENT BACTERIAL SIZE
 Most clinically relevant bacterial species range in size from
BACTERIA 0.25 - 1 μm in width and 1-3 μm in length
 Unicellular organisms that lack a nuclear membrane and  Bacterium is some hundred-fold larger than a virus, and ten-
true nucleus fold smaller than a eukaryotic cell
 Classified as prokaryotes (Greek: before kernel [nucleus]),  SIZE: eukaryotic cell > bacterium > virus
having no mitochondria, endoplasmic reticulum (ER), or 
Golgi bodies BACTERIAL SHAPE
 Common bacterial cellular morphologies include:
BACTERIAL MORPHOLOGY o Cocci – circular
 Vary in size, morphology, and cell-to-cell arrangements and o Coccobacilli – ovoid
in the chemical composition and structure of the cell wall o Bacillus – rod shaped
 Bacterial cell wall differences provide the basis for the Gram o Fusiform – tapered, pointed ends
Stain o Curved
 GRAM STAIN - the most fundamental test used in bacterial o Spiral – helical, like corkscrew.
identification.  Spirochetes – group of bacteria that is all
o Gram Stain Reagents helical.
 V – Crystal Violet, the Primary stain  Spirochetes vary in length and in the
 I – Gram’s ID, the Mordant (enhances the number of helical turns
stain; acts as bridge between the cell wall and o Pleomorphic – no defined shape
the stain)  Note: But not all helical bacteria are called spirochetes
 A – Ethyl Alcohol, the Decolorizer (thicker cell
wall in gram + enables the stain to stick) BACTERIAL ARRANGEMENT
 S – Safranin, the Secondary stain a) Pairs (DIPLO-)
b) Chains (STREPTO-)
c) Grape-like clusters (STAPHYLO-)
d) Group of four (TETRAD)
e) Packets of eight (SARCINAE)
f) Palisades (PALISADING)
g) Chinese characters (XVYL)

ALVAREZ RMT’26 3
 Throughout Heaven and Earth, I alone am the Honored One
BACTERIAL CELL STRUCTURE
CELL ENVELOPE
 Outermost structure, comprises:
o Outer membrane
 in gram-negative bacteria only
o Cell wall (MUREIN LAYER)
 composed of the
macromolecule
o Periplasm or Periplasmic Space
 in gram-negative bacteria only
o Cytoplasmic or cell membrane
 encloses the cytoplasm
 Gram negative: contains outer membrane, cell wall,
periplasmic space, and cell membrane
 Gram positive: contains cell wall and cell membrane only
OUTER MEMBRANE
 Found only in GRAM-NEGATIVE BACTERIA
 Function as the cell’s initial barrier to the environment
 Serve as primary permeability barriers to hydrophilic and
hydrophobic compounds and contain essential enzymes
and other proteins located in the periplasmic space
 Bilayered structure composed of lipopolysaccharide
o LIPOPOLYSACCHARIDE (responsible for the net
negative charge of gramnegative bacteria)
- Gives the surface of gramnegative bacteria a
net negative charge
 Proteins present in the outer membrane:
o PORINS – protein structures scattered throughout
the lipopolysaccharide macromolecules
 Water-filled structures that control the
passage of nutrients and other solutes,
including antibiotics, through the outer
membrane
 Number and types of porins vary with bacterial
species
 Influence the extent to which various
substances pass through the outer
membranes of different bacteria
 Passageway of different nutrients and other
solutes
o MUREIN LIPOPROTEINS – facilitate the attachment
of the outer membrane to the next internal layer in
the cell envelope, the cell wall
CELL WALL
 Peptidoglycan is thick in gram positive bacteria and thin in
gram negative bacteria
 Referred to as the Peptidoglycan or Murein Layer
 Gives the bacterial cell shape and strength to withstand
changes in environmental osmotic pressures that would
otherwise result in cell lysis
o Composition:
 A backbone composed of alternating sugar
components N-acetylglucosamine (NAG) and
N-acetylmuramic acid (NAM) connected by B
1-4 linkage
 Diaminopimelic acid (DAP) → unique element
of bacterial cell wall
o Some components responsible for pathogenicity:
 M-PROTEIN – present in Streptococcus
peptidoglycan pyogenes
 MYCOLIC ACID – present in Mycobacterium
tuberculosis
o Mycobacterium spp. have an unusual cell wall
structure:
 Cell wall contains N-glycolmuramic acid
instead of N-acetylmuramic acid
 Has a very HIGH LIPID CONTENT, which
creates hydrophobic permeability barrier (acid
fast staining)
o Different types of cell wall structures traditionally
have been categorized according to their staining
characteristics
 MAJOR TYPES OF CELL WALLS: Gram-Positive and
Gram-Negative Types
o Mycobacteria
 Stain gram-positive, have a modified cell wall
called an ACID-FAST CELL WALL
o Mycoplasmas and Ureaplasma
 Microorganisms that have no cell wall
ALVAREZ RMT’26 4
 Throughout Heaven and Earth, I alone am the Honored One

TYPES OF CELL WALL o BETA-HYDROXYMYRISTIC ACID → unique to

 gram positive cell wall Lipid A
 gram negative cell wall  LPS Functions:
 acid fast cell wall o Vital in evading the host defenses
 no cell wall o Contribute to the negative charge of the bacterial
surface, which stabilizes the membrane structure
 Gram Positive Cell Wall o Considered as an endotoxin
o composed of a very thick protective peptidoglycan
(murein) layer CHARACTERISTIC GRAM GRAM NEGATIVE
o consists of glycan (polysaccharide) chains of S POSITIVE
alternating N-acetyl-d-glucosamine (NAG) and Peptidoglycan Thick Thin
Nacetyl-d-muramic acid (NAM) Layer (Multilayered (Bilayered/Trilayered
o many antibiotics effective against gram-positive ) )
organisms (e.g., penicillin) act by preventing Teichoic Acids PRESENT ABSENT
synthesis of peptidoglycan IN MANY
o Types of Antibiotic: Periplasmic Space ABSENT PRESENT
 antibiotics that inhibit cell wall synthesis Outer Membrane ABSENT PRESENT
 antibiotics that inhibit protein synthesis LPS Content Virtually High
 antibiotics that inhibit nucleic acid synthesis None
NOTE: Gram-negative bacteria - thinner layer of Lipid and LPP Low High
peptidoglycan and a different cell wall structure, are less Flagellar Structure 2 rings in 4 rings in basal body
affected by these antibiotics basal body
Toxins produced EXOTOXIN ENDOTOXIN AND
 Other components of the gram-positive cell wall that EXOTOXIN
penetrate to the exterior of the cell are: Resitance to High Low
o TEICHOIC ACID Physical
 anchored to the peptidoglycan (N- Disruption
acetylmuramic acid) Cell Wall High Low
 glycerol or ribitol phosphate polymers combined Disruption by
with various sugars, amino acids, and amino Lysozyme
sugars Susceptibility to High Low
o LIPOTEICHOIC ACID Pen and
 anchored to the Plasma Membrane (PM) Sulfonamide
 linked to the next underlying layer, PM or Susceptibility to Low High
cellular membrane Strep. Chloram and
These two components are unique to the gram-positive cell Tetra
wall Inhibitions by High Low
Basic Dyes
o TEICHURONIC ACIDS Resistance to Low High
 Similar polymers, but the repeat units Anionic Detergents
include sugar acids (eg, N- Resistance to High Low
acetylmannosuronic or d-glucosuronic acid) Sodium Azide
instead of phosphoric acids Resistance to High Low
 synthesized in place of teichoic acids when Drying
phosphate is limiting
PROPER GRAM GRAM NEGATIVE
GRAM NEGATIVE CELL WALL TIES POSITIVE
 Composed of two layers: Shape Spherical, rod- Spherical, oval, straight or
o INNER PEPTIDOGLYCAN LAYER shape or curved, helical or
 much thinner than in gram-positive cell filamentous filamentous
walls
Metabolis CHEMOORGA PHOTOTROPHIC – use
o OUTER PEPTIDOGLYCAN LAYER
m NOHE light energy for certain
 Outside the peptidoglycan layer is an
TEROTROPHI metabolic functions
additional outer membrane
C – They CHEMOLITHOAUTOTROP
 contains proteins, phospholipids, and
require organic HIC – obtain the necessary
lipopolysaccharide (LPS)
substrate to get carbon for metabolic
its carbon for processes from carbon
THREE REGIONS OF LIPOPOLYSACCHARIDE (LPS) growth and dioxide in the environment
o ANTIGENIC O-SPECIFIC POLYSACCHARIDE development CHEMOORGANOHETERO
o CORE POLYSACCHARIDE TROPHIC
 ketodeoxyoctanoic acid (KDO)and Endospor PRESENT IN ABSENT
heptose e SOME
o INNER LIPID A GROUPS
 Also called ENDOTOXIN Reproduc Binary Fission Binary Fission
 Responsible for producing fever and tion
shock conditions in patients infected with
gram negative bacteria

ALVAREZ RMT’26 5
 Throughout Heaven and Earth, I alone am the Honored One

 Clinical Use of Cell Wall: GRAM STAINING o Tsukamurella

o Corynebacterium
REAGENTS (VIAS) GRAM + GRAM -  Cell wall without mycolic acid
1st Stain Crystal Stains Stains o Streptococcus
Violet PURPLE PURPLE o Actinomadura
Mordant Gram’s Remains Remains o Dermatophilus
Iodine PURPLE PURPLE o Nocardiopsis
Decolorizer 95% Remains Becomes o Oerskavia
Alcohol or PURPLE COLORLESS
Acetone NO CELL WALL
Counterstain Safranin Remains Stains PINK  lack a cell wall and contain STEROLS in their cell
PURPLE membranes
 lack the rigidity of the cell wall
 GRAM POSITIVE BACTERIA:  seen in various shapes microscopically
o Micrococcus, Staphylococcus, Streptococcus,  Example: Mycoplasma and Ureaplasma
Peptococcus, Peptostreptococcus, Sarcina,  NOTE: Gram-positive and gram-negative cells can lose
Bacillus, Corynebacterium, Erysipelothrix, Listeria, their cell walls and grow as L-forms in media
Mycobacterium, Nocardia, Actinomyces, supplemented with serum or sugar to prevent osmotic
Clostridium, Propionobacterium rupture of the cell membrane
 GRAM NEGATIVE BACTERIA:
o Branhamella, Neisseria, Veilonella, Acinetobacter, PERIPLASMIC SPACE
Aeromonas, Alcaligenes, Bordetella, Brucella,  typically found only in gram-negative bacteria
Enterobacteria, Francisella, Legionella, Pasteurella,  bounded by the internal surface of the outer membrane and
Pseudomonas, Vibrio, Fusobacterium, Bacteriodes the external surface of the cellular membrane
encompassing the thin peptidoglycan layer
ACID FAST CELL WALL  contains the murein layer, consists gel-like matrix
 have a gram-positive cell wall structure containing nutrient-binding proteins that assist in the
 contain a waxy layer of glycolipids and fatty acids (mycolic capture of nutrients from the environment
acid) bound to the exterior of the cell wall  contains several enzymes involved in the degradation of
 More than 60% of the cell wall is lipid macromolecules and detoxification of environmental
solutes, including antibiotics that enter through the outer
MYCOLIC ACID membrane
 major lipid component  Periplasmic space is absent in gram-positive bacteria
 strong “hydrophobic” molecule that forms a
 lipid shell around the organism and affects its permeability
 makes Mycobacterium spp. difficult to stain with the Gram
stain

MYCOBACTERIUM AND NOCARDIA

 stain a faint blue (gram-positive) color
 best stained with an acid-fast stain

ZIEHL- KINYOUN ACID NON-

NELSEEN (cold FAS ACID
(hot method) T FAST
method)
1ST CARBOLFU CARBOLFU Stain Stains
Stain CHSIN CHSIN s RED
RED CYTOPLASMIC (INNER) MEMBRANE
Mordant STEAN TERGITOL Rem Remains  present in both gram-positive and gram-negative bacteria
ains RED and is the deepest layer of the cell envelope
RED  consist of phospholipid bilayer, various proteins (70%),
Decolori HCl, Acid HCl, Acid Rem Become including a number of enzymes vital to cellular metabolism
zer Alcohol Alcohol ains s  serves as an additional osmotic barrier
RED COLOR  Absence of sterols
LESS
Counter Methylene Methylene Rem Stains  Exceptions: Mycoplasma
stain Blue Blue ains BLUE o incorporate sterols (e.g., cholesterol), into their
RED membranes when growing in sterol-containing
media
CLINICALLY RELEVANT AEROBIC
ACTINOMYCETES II. CYTOPLASMIC STRUCTURE
 Cell wall containing mycolic acid  RIBOSOMES
o Nocarida o site of protein biosynthesis and gives the cytoplasm
o Rhoococcus a granular structure
o Gordonia o Consist of RNA and proteins

ALVAREZ RMT’26 6
 Throughout Heaven and Earth, I alone am the Honored One
o 70S in size and separates into two subunits, 50S and
30S
 STREPTOMYCIN AND GENTAMYCIN
o attach to the 30S subunit and interfere with protein
synthesis
 ERYTHROMYCIN AND CHLORAMPHENICOL
o interfere with protein synthesis by attaching to the
50S subunit
GENOME
 Consist of a single, circular chromosome
 lacks nuclear membrane and mitotic apparatus
 Appears as diffused nucleoid or chromatin body that is
attached to a mesosome (sac-like structure)
 Consists of a single continuous circular molecule ranging in
size from 0.58 to almost 10 million base pair
 Exemptions:
o Borrelia burgdorferi and Streptomyces coelicolor
 Few bacteria have dissimilar chromosomes: Vibrio cholera
and Brucella melitensis
PLASMID
 Extrachromosomal, double-stranded element of DNA that
is associated with virulence
 Located in the cytoplasm and serve as a site for the genes
to code for antibiotic resistance and toxin production
 Not essential for bacterial growth so a bacterial cell may or
may not contain a plasmid
 Sometimes disappears during cell division and it can make
bacteria (mostly Gram-neg) pathogenic
TWO KINDS OF PLASMID:
 LARGE PLASMID
o Responsible for the production of B-lactamase that
provide resistance to Blactam antibiotics (penicilli
and oxacillin)
 SMALL PLASMID
o Resistant to tetracyclines and chloramphenicol
INCLUSIONS BODIES
 Serve as the energy source or food reserve of the bacteria
or as a reservoir of structural building blocks
 Composed mainly of polysaccharides, they lessen osmotic
pressure
 Examples:
o glycogen, cyanophysin granules,poly-
Bhydroxybutyrate granules, carboxysomes
(cyanobacteria, nitrifying bacteria and thiobacilli),
gas vacuoles (cyanobacteria, halobacterium and
thiothrix) and polyphosphate granules(volutin and
metachromatic granules)
TWO COMMON TYPES OF GRANULES:
 GLYCOGEN
o storage form of glucose
 PYROPHOSPHATE GRANULES
o storage form for inorganic phosphates.
o Source of phosphate for nucleic acid and
phospholipid synthesis
 Examples:
o METACHROMATIC/VOLUTIN/BABES - ERNST
GRANULES (Corynebacteiurm diphteriae)
o BIPOLAR BOODIES (Yersinia pestis)
o MUCH GRANULES (Mycobacterium tubercolosis)
FOOD RESERVES/ENERGY SORCE:
 Poly-B-hydroxybutyric acid (PHB)
o Lipid like compound consisting of chains of B-
hyroxybutyric acid units connected through ester
linkages
o Produced when the source of nitrogen, sulfur or
phosphorus is limited and there is excess carbon in
the medium
 PHB and Glycogen
o carbon source when protein and nucleic acid
synthesis are
o resumed
 Sulfur granules
o Hydrogen sulfide and thiosulfate
ENDOSPORES/ASEXUAL SPORES
 Small, dormant structures located inside the bacterial cell
 Aid in the survival of bacteria against external conditions
 Produced within vegetative cells of some Gram-pos
bacteria
 Composed of dipicolinic acid and calcium ions = CALCIUM
DIPICOLINATE
 Some locations could be a means of microscopically
identifying bacteria (ex: Clostridum tetani – lollipop-like
appearance bc endospore is located in the terminal part of
the cell)
o Responsible for perpetuation, but not multiplication
 Examples: Bacillus and Clostridium
TYPES OF SPORES ACCORDING TO LOCATION:
 Terminal spore – Clostridium tetani
 Subterminal spore – Clostridium botulinum (between
central and terminal)
 Central spore – Bacillus anthracis
PROPERTIES OF ENDOSPORES
 CORE
o spore protoplast
o contains a complete nucleus (chromosome), all of
the components of the protein-synthesizing
apparatus, and an energy-generating system based
on glycolysis
 SPORE WALL
o innermost layer surrounding the inner spore
membrane
o contains normal peptidoglycan and becomes the cell
wall of the germinating vegetative cell
 CORTEX
o thickest layer of the spore envelope
o contains an unusual type of peptidoglycan, with
many fewer cross-links than are found in cell wall
peptidoglycan
 COAT
o composed of a keratin-like protein containing many
intramolecular disulfide bonds
o Impermeability of this layer confers on spores their
relative resistance to antibacterial chemical agents
 EXOSPORIUM
o composed of proteins, lipids, and carbohydrates
o consists of a paracrystalline basal layer and a
hairlike outer region
CELLULAR APPENDAGES
 play a role in the mediation of infection and in laboratory
identification, varies among bacterial species and even
among strains within the same species
ALVAREZ RMT’26 7
 Throughout Heaven and Earth, I alone am the Honored One
GLYCOCALYX
 Outward complex of polysaccharide on the bacterial
surface and other cells
 Helps the bacteria to attach to the surface of the solid
objects or tissues
 Appears as a capsule or a slime layer
CAPSULE
 organized and is firmly attached to the cell wall
 immediately exterior to the murein layer of gram-positive
bacteria and the outer membrane of gram-negative bacteria
 Made up of polysaccharide polymers
 Exception: Poly-D-glutamic acid capsules of Bacillus
 anthracis and Bacillus Licheniformis
 Protects the bacteria (virulence factor) from the attacks of
human defense system since it resists phagocytosis and
desiccation
SLIME LAYER
 unorganized material that is loosely attached to the cell wall
 Made up of polysaccharide
 Can either inhibit phagocytosis or aid in the adherence of
the bacteria to the host tissue or synthetic implants
 Facilitates and maintains bacterial colonization of biologic
(e.g., teeth) and inanimate (e.g., prosthetic heart valves)
surfaces through the formation of biofilms Extracellular
Polymetric Substance (EPS)
 helps cells in a biofilm attach to their target environment and
to each other
FLAGELLA
 exterior protein filaments that rotate and cause bacteria to
be motile
 complex structures, mostly composed of the protein
flagellin, intricately embedded in the cell envelope
 plays an important role in survival and the ability of certain
bacteria to cause disease
 antigenic (H antigens), and some of the immune responses
to infection are directed against these proteins
 Gliding motility:
o Capnocytophaga, Cyanobacteria, Myxobacteria
ARRANGEMENT OF THE FLAGELLA:
 ATRICHOUS - without flagellum
 MONOTRICHOUS - single flagellum at one end
 AMPHITRICHOUS - single flagellum at both ends
 LOPHOTRICHOUS - tuff or group of flagella on one end or
both ends
 PERITRICHOUS - entire cell surface covered with flagella
 True motility and Brownian Movement are best observed
through the HANGING DROP METHOD
 True motility
o bacteria seem to be going in a definite direction
 Brownian movement
o bacteria bounce back and forth rapidly due to the
bombardment of molecules of water
 Taxis
o Movement of bacteria toward or away from a
particular stimulus
WAYS OF DEMONSTRATING MOTILITY IN THE LAB:
 Hanging Drop Method
 SIM
 Flagellar staining
 Serologic test
 Fluorescent Antibody Technique (FAT)
 Swarming Phenomenon
 Darkfield Microscopy
AXIAL FILAMENTS
 bundles of fibrils that arise at the ends of the cell beneath
an outer sheath and spiral around the cell
 anchored at one end of the spirochete
 have a structure similar to that of flagella
 Rotation of the filaments produces a movement of the outer
sheath that propels the spirochetes in a spiral motion
 Movement is similar to the way a corkscrew moves through
a cork
 SPIROCHETES
o group of bacteria that have unique structure and
motility
o move by means of AXIAL FILAMENTS OR
ENDOFLAGELLA
PILI (FIMBRIA)
 Hair-like, proteinaceous structures that extend from the cell
membrane into the external environment; some may be up
to 2 μm long
 Hair-like microfibrils usually produced by flagellated Gram-
negative bacteria observable by electron microscopy
 serve as adhesinsthat help bacteria attach to animal host
cell surfaces, often as the first step in establishing infection
 composed of structural protein subunits Pilins
 Two types: for attachment and reproduction
 Twitching Motility
o a pilus extends by the addition of subunits of pilin,
makes contact with a surface or another cell, and
then retracts (powerstroke) as the pilin subunits are
disassembled – grappling hook model
o Results in short, jerky, intermittent movements
o Example: Pseudomonas aeruginosa, Neisseria
gonorrheae, and some stains on E. coli
NUCLEUS: INFORMATION CENTRAL
 NUCLEUS
o Control center of the cell
o contains the cell’s genome/DNA/genetic material
o DNA dictates the cell on what it’s going to do and
how it’s going to do it
ALVAREZ RMT’26 8
 Throughout Heaven and Earth, I alone am the Honored One

 NUCLEAR ENVELOPE organelles such as lysosomes, or for export

o double membrane that encloses the nucleus, from the cell (secretion)
separating its contents from the cytoplasm
o perforated by pore structures (100 nm diameter)
 PORE COMPLEX
o intricate protein structure that lines each pore
o regulates the entry and exit of proteins and RNAs, as
well as large complexes of macromolecules
 NUCLEAR LAMINA
o netlike array of protein filaments that maintains the
shape of the nucleus by mechanically supporting the ENDOPLASMIC RETICULUM: BIOSYNTHETIC
nuclear envelope lines the nuclear side of the FACTORY
envelop
 Extensive network of membranes that accounts for more
 NUCLEAR MATRIX
than half the total membrane in many eukaryotic cells.
o Framework of protein fibers extending throughout
 Consists of a network of membranous tubules and sacs:
the nuclear interior
Cisternae (Latin cisterna, a reservoir for a liquid)
 CHROMOSOMES
o ER membrane separates the internal compartment
o Discrete units of organized DNA that carry the
of the ER (ER lumen or cisternal space) from the
genetic information
cytosol
o Each contains one long DNA molecule associated
with many proteins.
 Smooth ER
o When cells are ready to divide DNA condenses into
o Outer surface lacks ribosomes
structures known as chromosomes
o Synthesis of lipids, metabolism of carbohydrates,
 CHROMATIN
detoxification of drugs and poisons, and storage of
o Complex of DNA and proteins making up
calcium ions
chromosomes
 Rough ER
o Tangled spread out form of DNA
o Studded with ribosomes on the outer surface
 NUCLEOLUS
o Secrete proteins that are produced by ribosomes
o Mass of densely stained granules and fibers
adjoining part of the chromatin.
o Ribosomal RNA (rRNA) is synthesized from
instructions in the DNA
o Structure where ribosomes are made

GOLGI APPARATUS: SHIPPING AND RECEIVING

CENTER
 Consists of flattened membranous sacs: CISTERNAE
 Has distinct structural directionality

2 SIDES:
RIBOSOMES: PROTEIN FACTORIES
 CIS FACE
 Made of ribosomal RNA and protein that carry out protein
o Means “on the same side,” and usually located near
synthesis
the ER
 Ribosomes build proteins in two cytoplasmic locales o Transport vesicles move material from the ER to the
o FREE RIBOSOMES Golgi apparatus
 suspended in the cytosol  TRANS FACE
 proteins made on free ribosomes function within
o Means “on the opposite side” that gives rise to
the cytosol
vesicles that pinch off and travel to other sites
o BOUND RIBOSOMES
 attached to the outside of the endoplasmic
reticulum or nuclear envelope
LYSOSOMES: DIGESTIVE COMPARTMENTS
 make proteins that are destined for insertion  Membranous sac of hydrolytic enzymes used to hydrolyze
into membranes, for packaging within certain macromolecules

ALVAREZ RMT’26 9
 Throughout Heaven and Earth, I alone am the Honored One

 Enzymes work best in the acidic environment found in COMPONENTS

lysosomes
 Garbage collectors that takes in damaged or worn out cell
parts; have enzymes that breaks it down
o Excessive leakage from a large number of
lysosomes can destroy a cell by self-digestion
o Lysosomes also use their hydrolytic enzymes to
recycle the cell’s own organic material:
AUTHOPHAGY
VACUOLES: DIVERSE MAINTENANCE
COMPARTMENTS
 Large vesicles derived from the endoplasmic reticulum and
Golgi apparatus
 Selective in transporting solutes
THE EVOLUTIONARY ORIGINS OF MITOCHONDRIA
AND CHLOROPLASTS
ENDOSYMBIONT THEORY
 An early ancestor of eukaryotic cells engulfed an oxygen-
using non-photosynthetic prokaryotic cell
 Engulfed cell formed a relationship with the host cell in
which it was enclosed, becoming an endosymbiont (a cell
living within another cell).
MITOCHONDRIA: CHEMICAL ENERGY
CONVERSION
 Sites of cellular respiration, the metabolic process that uses
oxygen to drive the generation of ATP by extracting energy
from sugars, fats, and other fuels
o CRISTAE
 Infoldings of the inner membrane
o INTERMEMBRANE SPACE
 Narrow region between the inner and outer
membranes
o MITOCHONDRIAL MATRIX
 Enclosed by the inner membrane.
 Contains many different enzymes as well as the
mitochondrial DNA and ribosomes
 MICROTUBULES - thickest of the three types
 MICROFILAMENTS - aka actin filaments; thinnest
 INTERMIDIATE FILAMENTS - fibers with diameters in a
middle range
MICROTUBULES
 Hollow rods constructed from a globular protein called
TUBULIN
 Each tubulin protein is a dimer: α-tubulin and β-tubulin.
 Shape and support the cell and also serve as tracks along
which organelles equipped with motor proteins can move
 Guide vesicles from the ER to the Golgi apparatus and from
the Golgi to the plasma membrane
 Involved in the separation of chromosomes during cell
division
CENTROSOMES AND CENTRIOLES
 CENTROSOME
o Region that is often located near the nucleus
o These microtubules function as compression-
resisting girders of the cytoskeleton
 CENTRIOLES
o Located within the centrosome
o Composed of nine sets of triplet microtubules
arranged in a ring
 CILIA AND FLAGELLA
o Specialized arrangement of microtubules is
responsible for the beating of flagella and cilia
microtubule-containing extensions that project from
some

PEROXISOMES: OXIDATION
 Specialized metabolic compartment bounded by a single
membrane
 Contain enzymes that remove hydrogen atoms from
various substrates and transfer them to oxygen (O2),
producing hydrogen peroxide (H2O2) as a by-product
 Use oxygen to break fatty acids down
 Detoxify alcohol in the liver and other harmful compounds

CYTOSKELETON: SUPPORT AND MOTILITY

 Give mechanical support to the cell and maintain its shape
 Provides anchorage for many organelles and even cytosolic
enzyme molecules.
 Involve in some types of cell motility

ALVAREZ RMT’26 10
 Throughout Heaven and Earth, I alone am the Honored One

MICROFILAMENTS (ACTIN FILAMENTS)

 Thin solid rods
 Built from molecules of actin a globular protein
 Bears tension (pulling forces)
 Three-dimensional network formed by microfilaments just
inside the plasma membrane (cortical microfilaments) helps
support the cell’s shape.
 Role in cell motility
 Contraction of muscle cells: ACTIN and MYOSIN

INTERMEDIATE FILAMENTS
 Diameter larger than the diameter of microfilaments but
smaller than that of microtubules
 More permanent fixtures of cells than are microfilaments
and microtubules, which are often disassembled and
reassembled in various parts of a cell

CELL CYCLE OF PROKARYOTES (BINARY

FISSION):
 Chromosomes are replicated (single-circularly arranged
chromosome)
 The identical copies are attached to the plasma membrane
 Cell elongates and forms new cell wall in between the two
chromosomes which separates them

 RESULT: 2 cells with identical copy of chromosome

 Difference with eukaryotes: bacteria do not need to
dissolve nuclear membrane or assemble mitotic spindle
 Note: each bacterium can divide every 20 mins. They
can produce billions in a matter of 10 hours

SIMILARITY OF CELL CYCLE OF EUKARYOTES TO

 CELL ENVELOPE STRUCTURES PROKARYOTES:
 PLASMA MEMBRANE  DNA replication
 Phospholipid bilayer with embedded proteins that envelops  Copy separation
the cytoplasm and regulates transport of macromolecules  Cytoplasm division
into and out of the cell  *more complex on eukaryotes
 Contains a substantial amount of cholesterol  NOTE: Each eukaryotic specie has a characteristic
 Presence of STEROLS number of chromosomes per cell
 CELL WALL o Ex: humans has 46 chromosomes – 2 sets of
 Provide rigidity and strength to the exterior of the cell chromosomes (consequence of sexual
 Most eukaryotic cells do not have cell walls reproduction): 1 set from male parent and 1 set from
 Fungi have cell walls principally made of polysaccharides female parent – together consisting of homologous
pair – bc each chromosome in 1 set contains a
THE CELL CYCLE complementary in the other set
 Interphase – where cells spends most of its time: where o There are 23 homologous pairs
cell growth primarily occurs  22 pair of autosomes (alike in structure and
o G1 – longest phase; produce extra organelles and size)
proteoins  1 pair of sex chromosomes (not identical)
o S phase (synthesis) – DNA replication  Diploid – 2 sets of genetic information
o G2 – prep for mitosis o Describes a cell that contain two copies of each
 Mitosis – where active cell division occurs (M phase) chromosome
o Prophase o Somatic cells
o Prometaphase  Not all eukaryotic cells are diploid
o Metaphase  Haploid – 1 set of genetic information; reproductive cells
o Anaphase o Egg cells, sperm cells, spores
o Telophase
3 ESSENTIAL ELEMENTS OF FUNCTIONAL
CHROMOSOMES
 CENTROMERE – the attachment part of the spindle
microtubules
 It is a constricted region that stain less strongly than the rest
of the chromosomes
o spindle microtubules – filaments responsible for
moving chromosomes during cell division
 Chromosomes are classified based on the
ALVAREZ RMT’26 11
 Throughout Heaven and Earth, I alone am the Honored One
 POSITION OF CENTROMERE:
 Metacentric – gitna
o Sub metacentric – medyo taas ng konti
o Acrocentric chromosomes – medyo taas
o Telocentric chromosomes – dulo
o P stands for short arm
o Q stands for long arm
 TELOMERES – natural ends or tips of linear chromosome
 Function: stabilizes chromosome
 ORIGIN OF REPLICATION – where the DNA synthesis
begin
CELL CYCLE
 Sequence of activities as a cell prepares for division and
then divides
2 MAJOR STAGES:
 INTERPHASE (not dividing)
o Composed of G1, S phase, G1, and G0 in some
cases
 MITOTIC PHASE/M PHASE (dividing)
o Composed of prophase, prometaphase, metaphase,
anaphase, and telophase
DIFFERENCE OF MEIOSIS AND MITOSIS (ACTIVE
CELL DIVISIONS):
 MITOSIS – division of non-sex cells; SOMATIC CELLS
o Cell duplicates its chromosomes, then apportions
one set into each of two resulting daughter cells
 MEIOSIS – division of sex cells; SEX CELLS/GERM
CELLS/NONSOMATIC CELLS
o Half the amount of genetic material in somatic cells
FUNCTIONS OF CELL DIVISION
 IN PROKARYOTIC CELLS: for reproduction (asexual
reproduction); BINARY FISSION
 IN EUKARYOTIC CELLS: for renewal or repair - normal
wear and tear or accidents; MITOSIS
 Growth, development, maintaining health, and healing from
 disease or injury require an intricate interplay between the
rates of these two processes:
o MITOSIS gives rise to two somatic cells from one
o APOPTOSIS form of cell death/ program cell death
 Greek for “leaves falling from a tree,” is a
precise, genetically programmed sequence of
events that is a normal part of development
PHASES OF THE CELL CYCLE
 Mitosis is just one part of the cell cycle.
 In fact, the mitotic (M) phase, *10% which includes both
mitosis and cytokinesis, is usually the SHORTEST PHASE
of the cell cycle.
 The mitotic phase alternates with a much longer stage
called INTERPHASE, which often accounts for about 90%
of the cycle.
o Interphase can be divided into subphases: the G1
PHASE (“first gap”), the S PHASE (“synthesis” -DNA
synthesis), and the G2 PHASE (“second gap”).
 G1 growth of cell
 S production of DNA and some substances such as
proteins and carbohydrates
 G2 additional metabolic activities
 Checkpoints – occurs before the next phase; (ex:
checkpoint in G1 checks if the size of a certain cell is correct
before it proceeds to the S phase)
o The G PHASE were misnamed as “gaps” when they
were first observed because the cells appeared
inactive, but we now know that intense metabolic
activity and growth occur throughout interphase.
INTERPHASE
 It is the extended period of growth and development
between cell divisions.
 Although little activity can be observed with a light
microscope, the cell is quite busy: DNA is being
synthesized, RNA and proteins are being produced, and
hundreds of biochemical reactions necessary for cellular
functions are taking place.
 In addition to growth and development, interphase includes
several CHECKPOINTS, which regulate the cell cycle by
allowing or prohibiting the cell’s division.
o These checkpoints, like the checkpoints in the M
phase, ensure that all cellular components are
present and in good working order before the cell
proceeds to the next stage.
o Checkpoints are necessary to prevent cells with
damaged or missing chromosomes from
proliferating.
o Defects in checkpoints can lead to unregulated cell
growth, as is seen in some cancers.
o By convention, interphase is divided into three
subphases: G1, S, and G2.
3 SUBPHASES OF INTERPHASE:
 G1
o Interphase begins with G1 (for gap 1).
o In G1, the CELL GROWS, and proteins, (lipids and
carbohydrates - imporatant in extraplasma
membrane) necessary for cell division are
synthesized; this phase typically lasts several hours.
ALVAREZ RMT’26 12
 Throughout Heaven and Earth, I alone am the Honored One
o Near the end of G1, a critical point termed the G1/S
CHECKPOINT holds the cell in G1 until the cell has
all of the enzymes necessary for the replication of
DNA.
o After this checkpoint has been passed, THE CELL
IS NOW COMITTED TO DIVIDE
o Before reaching the G1/S checkpoint, cells may exit
from the active cell cycle in response to regulatory
signals and pass into a nondividing phase called G0,
which is a stable state during which cells usually
maintain a constant size.
 A cell in G0 maintains its specialized
characteristics but does not replicate its DNA or
divide.
 From G0 , a cell may also proceed to mitosis
and divide, or die.
 Apoptosis may ensue if the cell’s DNA is so
damaged that cancer might result.
 G0, then, is when a cell’s fate is either decided
or put on hold.
 They can remain in G0 for an extended length
of time, even indefinitely, or they REENTER
G1and the active cell cycle.
 Many cells never enter G0; rather, they cycle
continuously.
 Examples of cells that goes in G0: cells in the liver, nerve
cells and heart cells. This is because once they reach
maturity, nerve and heart cells do not divide again, so they
stay in the G0 phase.
 Note: CELLS IN THE EMBYRO MAY SKIP G1 CELLS IN
THE BONE MARROW SPEED UP THROUGH G1 IN 16-
24 HOURS
S PHASE
 After G1, the cell enters the S phase (for DNA
SYNTHESIS), in which each chromosome duplicates.
 Although the cell is committed to divide after the G1/S
checkpoint has been passed, DNA synthesis must take
place before the cell can proceed to mitosis.
 If DNA synthesis is blocked (by drugs or by a mutation), the
cell will not be able to undergo mitosis.
 Before the S phase, each chromosome is unreplicated;
after the S phase, each chromosome is composed of two
chromatids.
 The basis in counting chromosomes is the number of
centromeres
o During S phase, the cell replicates its entire genome.
o As a result, each chromosome then consists of two
copies joined at an area called the CENTROMERE
o In most human cells, S phase takes 8-10 HOURS
o Many proteins are also synthesized during this
phase, including those that form the MITOTIC
SPINDLE that will pull the chromosomes apart.
G2
 After the S phase, the cell enters G2 (gap 2)
 In this phase, several additional biochemical events
necessary for cell division take place.
 G2 TAKES PLACE BEFORE MITOSIS BUT AFTER THE
DNA HAS BEEN REPLICATED
 More proteins are synthesized during this phase.
 Membranes are assembled from molecules made during
G1 and are stored as small, empty vesicles beneath the
plasma membrane.
 These vesicles will merge with the plasma membrane to
enclose the two daughter cells.
 The important G2/ M CHECKPOINT is reached near the
end of G2.
 This checkpoint is passed only if the cell’s DNA is
undamaged.
 Damaged DNA can inhibit the activation of some proteins
that are necessary for mitosis to take place.
 After the G2/M checkpoint has been passed, the cell is
ready to divide and enters the M phase.
 Although the length of interphase varies from cell type to
cell type, a typical dividing mammalian cell spends
 about 10 hours in G1, 9 hours in S, and 4 hours in G2.
MITOSIS
 As mitosis begins, the replicated chromosomes are
condensed enough to be visible, when stained, under a
microscope.
 The two long strands of identical chromosomal material in
a replicated chromosome are called chromatids.
 At a certain point during mitosis, a replicated chromosome’s
centromere splits, allowing its chromatid pair to separate
into two individual chromosomes. (Although the centromere
of a replicated chromosome appears as a constriction, its
DNA is replicated.)
 Mitosis occurs in both haploid and diploid cells.

 Mitosis is conventionally broken down into FIVE STAGES:
o PROPHASE
o PROMETAPHASE
o METAPHASE
o ANAPHASE, and
o TELOPHASE
o
ALVAREZ RMT’26 13
 Throughout Heaven and Earth, I alone am the Honored One
 Overlapping with the latter stages of mitosis,
cytokinesis completes the mitotic phase.
5 STAGES OF MITOSIS:
PROPHASE (CONDENSATION OF CHROMOSOME)
 In the G2 stage of the cell cycle, just prior to the start of M,
each chromosome consists of two sister chromatids, and
the centrioles have duplicated to produce two pairs.
 In prophase, the CHROMATIN CONDENSES so they
gradually appear shorter and fatter under the microscope.
 By late prophase, each chromosome, which was duplicated
during the preceding S phase of interphase, can be seen to
consist of two sister chromatids.
 While condensation is occurring, the NUCLEOLUS
SHRINKS and eventually disappears in most species.
 Many mitotic events depend on the mitotic spindle (spindle
apparatus), a structure consisting of fibers composed of
microtubules made of special proteins called tubulins.
 The mitotic spindle assembles outside the nucleus
duringprophase.
 In most animal cells, the centrioles are the focal pointsfor
spindle assembly; higher plant cells usually lack centrioles,
but they do have a mitotic spindle.
 Then, during mitosis, each new centriole pair becomes the
focus of a radial array of microtubules called the aster.
 Prometaphase (DISINTEGRATION OF NUCLEAR
MEMBRANE)
 The nuclear envelope breaks down at the end of prophase,
denoting the beginning of prometaphase.
 THE DISINTEGRATION OF THE NUCLEAR MEMBRANE
MARKS THE START OF PROMETAPHASE
 The developing spindle now enters the former nuclear area.
 A specialized multiprotein complex called a kinetochore
binds to each centromere.
 The kinetochores are the sites for the attachment of the
chromosomes to spindle microtubules known as
KINETOCHORE MICROTUBULES
 Nonkinetochore microtubules—spindle microtubules that
do not bind to kinetochores—also originate from each
spindle pole and overlap in the middle of the spindle
METAPHASE (CENTROMERE ALIGN IN THE
MIDDLE)
 During metaphase, the kinetochore microtubules orient the
chromosomes so that their centromeres become aligned at
the METAPHASE PLATE, a plane halfway between the two
spindle poles, with the long axes of the chromosomes
oriented at 90 degrees to the spindle axis.
 The centrosomes, now at opposite ends of the cell with
microtubules radiating outward and meeting in the middle
of the cell, center at the spindle poles.
 A SPINDLE-ASSEMBLY CHECKPOINT ensures that each
chromosome is aligned on the metaphase plate and
attached to spindle fibers from opposite poles.
 It had been accepted for many years that metaphase
chromosomes are the most condensed form of
chromosomes in mitosis (and meiosis).
 Recently, examination of 3D reconstructions of microscope
images of living mammalian cells taken with high-power
microscopes has revealed that further chromosome
condensation occurs just after the chromosomes have
finished separating in the subsequent anaphase stage. This
late condensation serves to minimize the potential problem
of chromosome arms extending over the plane of division,
which could result in mechanical damage to the
chromosomes.
ANAPHASE (PULLING AWAY OF CHROMATIDS)
 Anaphase begins suddenly when the COHESINE holding
together the sister chromatids of each chromosome are
cleaved by an enzyme called SEPARASE
 -ase for enzyme
 -ine for protein
 Once separated, the chromatids become full-fledged
chromosomes that move toward opposite ends of the cell.
 During anaphase, the joined centromeres of sister
chromatids separate, giving rise to two daughter
chromosomes.
 Once the paired kinetochores on each chromosome
separate, the sister chromatid pairs undergo disjunction
(separation), and the daughter chromosomes move toward
the opposite poles.
 In anaphase, the daughter chromosomes are pulled toward
the opposite poles of the cell by the shortening microtubules
attached to the kinetochores.
 The microtubules that connect the chromosomes to the
spindle poles are composed of subunits of a protein called
TUBULINE
 Chromosome movement is due to the disassembly of
tubulin molecules at both the kinetochore end (called
 the end) and the spindle end (called the end) of the spindle
fiber.
 Special proteins called MOLECULAR MOTORS
disassemble tubulin molecules from the spindle and
generate forces that pull the chromosome toward the
spindle pole.
 ANAPHASE - shortest stage of mitosis
ALVAREZ RMT’26 14
 Throughout Heaven and Earth, I alone am the Honored One
TELOPHASE (NUCLEAR ENVELOPE REFORMS)
 At the start of telophase, the two sets of daughter
chromosomes are assembled into two groups at opposite
ends of the cell.
 The chromosomes begin to uncoil and assume the
elongated state characteristic of interphase.
 A nuclear envelope forms around each group of
chromosomes, the spindle microtubules disappear, and the
nucleolus or nucleoli reform.
 At this point, nuclear division is complete and the cell now
has TWO NUCLEI.
CYTOKINESIS
 Cytokinesis is DIVISION OF CYTOPLASM; usually, it
follows the nuclear division stage of mitosis and is
completed by the end of telophase.
 Cytokinesis compartmentalizes the two new nuclei into
separate daughter cells, completing mitosis and cell. In
animal cells, cytokinesis occurs by a process known as
CLEAVAGE
 The first sign of cleavage is the appearance of a
CLEAVAGE FURROW, a shallow groove in the cell surface
near the old metaphase plate.
 On the cytoplasmic side of the furrow is a contractile ring of
actin microfilaments associated with molecules of the
protein myosin.
 The actin microfilaments interact with the myosin
molecules, causing the ring to contract.
 The contraction of the dividing cell’s ring of microfilaments
is like the pulling of a drawstring. The cleavage furrow
deepens until the parent cell is pinched in two, producing
two completely separated cells, each with its own nucleus
and its own share of cytosol, organelles, and other
subcellular structures.
GENETIC CONSEQUENCES OF THE CELL CYCLE
 From a single cell, the cell cycle produces two cells
thatcontain the same genetic instructions.
 The resulting daughter cells are genetically identical with
each other and with their parent cell because DNA
synthesis in the S phase creates an exact copy of each
DNA molecule, giving rise to two genetically identical sister
chromatids.
 Mitosis then ensures that one of the two sister chromatids
from each replicated chromosome passes into each new
cell.
 Another genetically important result of the cell cycle is that
each of the cells produced contains a full
 complement of chromosomes: there is no net reduction or
increase in chromosome number.
 Not all cells resulting from the cell cycle are identical in their
cytoplasmic content.
THE CELL CYCLE CLOCK: CYCLINS AND CYCLIN-
DEPENDENT KINASES
 Rhythmic fluctuations in the abundance and activity ofcell
cycle control molecules pace the sequential events of the
cell cycle.
 These regulatory molecules are mainly proteins of two
types: protein kinases and cyclins.
PROTEIN KINASES
 Are enzymes that activate or inactivate other proteins by
phosphorylating them.
 Many of the kinases that drive the cell cycle are actually
present at a constant concentration in the growing cell, but
much of the time they are in an inactive form.
 To be active, such a kinase must be attached to a cyclin, a
protein that gets its name from its cyclically fluctuating
concentration in the cell.
o Cyclin-dependent kinases – cannot function without
cyclin; there should be a cyclin that will bind to it to
activate it
o Because of this requirement, these kinases are
called cyclin-dependent kinases, or CDKS.
o CDKs + Cyclin = MPF (maturation promoting
factor/m-phase promoting factor)
o The activity of a Cdk rises and falls with changes in
the concentration of its cyclin partner.
 MPF, the cyclin-CDK complex, activity correspond to the
peaks of cyclin concentration.
 The cyclin level rises during the S and G2 phases and then
falls abruptly during M phase.
 The initials MPF stand for “maturation-promoting factor,”
but we can think of MPF as “M-phase-promoting factor”
because it triggers the cell’s passage into the M phase, past
the G2 checkpoint.
 NOTE:
 MPF is for cyclin-dependent kinases, or CDKs CDKs -
activates different kinases that initiates proteins in S
phase. MPF - aids in nuclear membrane breakdown,
spindle formation, and chromosome condensation.
Important in regulation and processing of cell cycle.
There is a peak of MPF activity during mitotic phase
because of cyclin.
ALVAREZ RMT’26 15
 Throughout Heaven and Earth, I alone am the Honored One
 When cyclins that accumulate during G2 associate with
CDK molecules, the resulting MPF complex phosphorylates
a variety of proteins, initiating mitosis.
 MPF acts both directly as a kinase and indirectly by
activating other kinases.
 During anaphase, MPF helps switch itself off by initiating a
process that leads to the destruction of its own cyclin.
 The noncyclin part of MPF, the CDK, persists in the cell,
inactive until it becomes part of MPF again by associating
with new cyclin molecules synthesized during the S and G2
phases of the next round of the cycle.
CYCLINS
 Proteins known as cyclins (named because their
concentration increases and decreases in a regular pattern
through the cell cycle) and enzymes known as cyclin-
dependent kinases (Cdks) are the key components in the
regulatory events that occur at checkpoints.
 At the G1-to-S checkpoint, two different G1 cyclin–Cdk
complexes form, resulting in activation of the kinases.
 The kinases *are enzymes catalyze a series of
phosphorylations (addition of phosphate groups) of cell
cycle control proteins, affecting the functions of those
proteins and leading, therefore, to transition into the S
phase.
o Important in our checkpoints
 A similar process occurs at the G2-to-M checkpoint.
 A cyclin binds to a Cdk to form a complex.
 Until the cell is ready to enter mitosis, phosphorylation of
the Cdk by another kinase keeps the Cdk inactive.
 At that time, a phosphatase removes the key phosphate
from the Cdk, activating the enzyme.
 Phosphorylations of proteins by the Cdk move the cell into
mitosis.
**Without MPF, they cannot enter mitosis
STOP AND GO SIGNS: INTERNAL AND EXTERNAL
SIGNALS AT THE CHECKPOINTS
 Animal cells generally have built-in stop signals that halt the
cell cycle at checkpoints until overridden by go-ahead
signals. (The signals are transmitted within the cell by the
kinds of signal transduction pathways)
 Many signals registered at checkpoints come from cellular
surveillance mechanisms inside the cell.
 These signals report whether crucial cellular processes that
should have occurred by that point have in fact been
completed correctly and thus whether or not the cell cycle
should proceed.
 Checkpoints also register signals from outside the cell.
 Three important checkpoints are those in G1, G2, and
M phases.
o G1/S CHECKPOINT – if you passed, cell is
committed to divide
o G2/M CHECKPOINT
 For many cells, the G1 checkpoint—dubbed the “restriction
point” in mammalian cells—seems to be the most
important.
 *Requirement: enough proteins, lipids, and
carbohydrates, and the size of the cell is enough
 If a cell receives a go-ahead signal at the G1 checkpoint, it
will usually complete the G1, S, G2, and M phases and
divide.
 If it does not receive a go-ahead signal at that point, it may
exit the cycle, switching into a nondividing state called the
G0 phase.
 Most cells of the human body are actually in the G0 phase.
 Mature NERVE CELLS AND MUSCLE CELLS never
divide.
 Other cells, such as liver cells, can be “called back” from
the G0 phase to the cell cycle by external cues, such as
growth factors released during injury.
LOSS OF CELL CYCLE CONTROLS IN CANCER
CELLS
 CANCER - Uncontrolled growth of cells
 Cancer cells do not need the normal signals that regulate
the cell cycle.
 In culture, they do not stop dividing when growth factors are
depleted.
 A logical hypothesis is that cancer cells do not need growth
factors in their culture medium to grow and divide.
 They may make a required growth factor themselves, or
they may have an abnormality in the signaling pathway that
conveys the growth factor’s signal to the cell cycle control
system even in the absence of that factor.
 Another possibility is an abnormal cell cycle control system.
 In these scenarios, the underlying basis of the abnormality
is almost always a change in one or more genes (for
example, a mutation) that alters the function of their protein
products, resulting in faulty cell cycle control.
 If and when they stop dividing, cancer cells do so at random
points in the cycle, rather than at the normal
 checkpoints. Cancer cells can go on dividing indefinitely in
culture if they are given a continual supply of nutrients; in
essence, they are “immortal.”
 A striking example is a cell line that has been reproducing
in culture since 1951, called HeLa cells because their
original source was a tumor removed from a woman named
Henrietta Lacks.
 Cells in culture that acquire the ability to divide indefinitely
are said to have undergone transformation, the process that
causes them to behave like cancer cells.
 By contrast, nearly all normal, nontransformed mammalian
cells growing in culture divide only about 20 to 50 times
before they stop dividing, age, and die.
 The abnormal cells may remain at the original site if they
have too few genetic and cellular changes to survive at
ALVAREZ RMT’26 16
 Throughout Heaven and Earth, I alone am the Honored One

another site. In that case, the tumor is called a BENIGN  In most cases, the divisions are accompanied by
TUMOR. cytokinesis, so the meiosis of a single diploid cell produces
 Most benign tumors do not cause serious problems and can HAPLOID CELLS (4 HAPLOID CELLS)
be removed by surgery.
 In contrast, a MALIGNANT TUMOR includes cells whose
genetic and cellular changes enable them to spread to new
tissues and impair the functions of one or more organs;
these cells are also considered “transformed cells.”
 METASTASIS – the process by which cancer cells spread
to other parts of the body

MEIOSIS
 aka reduction division
 formation of sex cells/gametes
 MITOSIS is similar to MEIOSIS, particularly MEIOSIS II
 From 46 chromosomes to 23 chromosomes
 From diploid to haploid
 From 2n to 4n

OVERVIEW:
 Interphase - cell growth, DNA replication, cellular activities
 PPMAT I (Homologous Pairs)/ Meiosis I - Homologous
pairs
 PROPHASE I
o Leptonema – condensation of chromatids
o Zygonema – they line up with their homologous pairs
o Pachynema – crossing over (it is the transfer of
genetic information and exchange it with each other,
that results to recombinant chromosomes)
o Diplonema – synaptonemal complex begin to move
apart
 PROMETAPHASE I - Nuclear envelop disappears,
microtubules enter to prometaphase I
 METAPHASE I - Homologous pairs align in the middle
 ANAPHASE I - Homologous pairs are pulled away by
spindle fibers
 TELOPHASE I - Production of two new cells
 CYTOKINESIS – cell division
 PPMAT II (Sister chromatids)/Meiosis II - Sister
chromatids; NO crossing over, NO pairing/line up of
homologous pair
MEIOSIS
 Meiosis is the two successive divisions of a DIPLOID
nucleus after only one DNA replication (chromosome
duplication) cycle.
 The original diploid nucleus contains one haploid set of
chromosomes from the mother and one set from the father.
 It results in the formation of haploid gametes (eggs and
sperm by gametogenesis);
 * From 2n to 4n (4n is the resulting cell)
 Before meiosis, the DNA that makes up homologous
chromosomes replicates, and during meiosis these
chromosomes pair and then undergo two divisionsmeiosis
I and meiosis II—each consisting of a series of stages.
 Note:
o Meiosis I – REDUCTIONAL DIVISION
o Meiosis II - EQUATIONAL DIVISION, similar to
mitosis
 MEIOSIS I results in a reduction in the number of
chromosomes in each cell from diploid to haploid
(reductional division—each resulting pair of attached sister
chromatids counts as a single chromosome)
 MEIOSIS II results in the separation of the sister
chromatids.
MEIOSIS I: THE FIRST MEIOTIC DIVISION
 Meiosis I, in which the chromosome number is reduced
from diploid to haploid, consists of FIVE
 STAGES:
o Prophase, prometaphase, metaphase, anaphase,
telophase
PROPHASE I
 When prophase I begins, the chromosomes have already
duplicated, with each consisting of two sister chromatids
attached at centromere.
 Prophase I is divided into a number of substages.
 Prophase I of meiosis is similar to prophase of
mitosis.(there’s condensation)
 The important difference between prophase I of meiosis
and prophase of mitosis is that HOMOLOGOUS
CHROMOSOMES PAIR WITH EACH OTHER in meiosis,
and CROSSING-OVER OCCURS only in meiosis.
 LEPTONEMA (condensation of chromatids)
o In leptonema (early prophase I, the LEPTOTENE
STAGE) the extended chromosomes begin to
condense and become visible as long, thin threads.
o Once a cell enters leptonema, it is committed to the
meiotic process.
 ZYGONEMA (line up with their homologous pairs)
o In zygonema (early to middle prophase I,
ZYGOTENE STAGE), the chromosomes continue to
condense.
o The homologous pairs of chromosomes actively find
each other and align roughly along their lengths.

ALVAREZ RMT’26 17
 Throughout Heaven and Earth, I alone am the Honored One

o Each pair of homologs then undergoes

SYNAPSIS—the formation along the length of the
chromatids of a zipperlike structure called the
SYNAPTONEMAL COMPLEX which aligns the two
homologs precisely, base pair for base pair.
 Synaptonemal complex – fibers between homologous
pairs; results to synapsis resulting to crossing over

 PACHYNEMA (crossing over)

o Pachynema (middle prophase I, the PACHYTENE)
starts when synapsis is completed.
o Because of the replication that occurred earlier, each
synapsed set of homologous chromosomes consists
of four chromatids and is called a bivalent or a tetrad.
o During pachynema, a most significant event for
genetics occurs: CROSSING OVER - the reciprocal
physical exchange of chromosome segments at
corresponding positions along pairs of homologous
chromosomes.
o The positions at which crossing-over occur along the
chromosomes are largely random, and vary from
one meiosis to another.
o The physical exchange that occurs in crossing-over
is facilitated by the alignment of the homologous
chromosomes brought about by the synaptonemal
complex.
o If there are genetic differences between the
homologs, crossing-over can produce new gene
combinations in a chromatid.
 Because of this crossing over along with
independent assortment and random
fertilization, we have GENETIC VARIATION
among offsprings.
o There is usually no loss or addition of genetic
material to either chromosome, since crossing-over
involves reciprocal exchanges.
o A chromosome that emerges from meiosis with a
combination of alleles that differs from the
combination with which it started is called a
RECOMBINANT CHROMOSOME
o Therefore, crossing-over is a mechanism that can
give rise to genetic recombination.
o At the end of pachynema, the synaptonemal
complex is disassembled, and the chromosomes
have started to elongate.

 DIPLONEMA (synaptonemal complex begin to move

apart)
o In diplonema (middle to late prophase I, the
DIPLOTENE) the synaptonemal complex
disassembles and the homologous chromosomes
begin to move apart.The result of crossing-over
becomes visible during diplonema as a cross-
shaped structure called a chiasma (plural,
chiasmata).
o At each chiasma, the homologous chromosomes are
very tightly associated.
o Because all four chromatids may be involved in
crossingover events along the length of the
homologs, the chiasma pattern at this stage may be
quite complex.
 DIAKINESIS
o (late prophase I), the chromosomes condense even
more, making it now possible to see the four
members of the tetrads.
o The chiasmata are clearly visible at this stage.

ALVAREZ RMT’26 18
 Throughout Heaven and Earth, I alone am the Honored One
II. PROMETAPHASE I
 In prometaphase I, the NUCLEOLI DISAPPEARS, the
nuclear envelope breaks down, and the meiotic spindle that
has been forming between the separating centriole pairs
enters the former nuclear area.
 As in mitosis, kinetochore microtubules attach to the
chromosomes; that is, kinetochore microtubules from one
pole attach to both sister kinetochores of one duplicated
chromosome, and kineteochore microtubules from the
other pole attach to both sister kinetochores of the other
duplicated chromosome in a tetrad.
 Nonkinetochore microtubules from each pole overlap in the
center of the cell.
III. METAPHASE I (MIDDLE)
 In metaphase I, the kinetochore microtubules align the
tetrads on the METAPHASE PLATE
 Importantly, the pairs of homologs (the tetrads) are found at
the metaphase plate.
 In contrast, in mitosis, replicated homologous
chromosomes (sister chromatid pairs) align independently
of one another at the metaphase plate.
IV. ANAPHASE I (AWAY)
 At this time, homologous chromosomes have segregated
from each other, but sister chromatids remain attached at
their respective centromeres.
 In other words, a key difference between meiosis I and
mitosis is that sister chromatids remain joined after
metaphase in meiosis I, whereas they separate in mitosis.
V. TELOPHASE I
 In telophase I, the dyads complete their migration to
opposite poles of the cell and the spindle disassembles.
 In some species, but not all, new nuclear envelopes form
around each haploid grouping.
 In most species, cytokinesis follows telophase I, producing
two haploid cells.
 Thus, meiosis I, which begins with a diploid cell that
contains one maternally derived and one paternally derived
set of chromosomes, ends with two nuclei, each of which is
haploid and contains one mixed-parental set of dyads.
 After cytokinesis, each of the two progeny cells has a
nucleus with a haploid set of dyads.
MEIOSIS II: THE SECOND MEIOTIC DIVISION
 No DNA replication occurs between meiosis I and meiosis
II.
 Meiosis II is similar to a mitotic division.
I. PROPHASE II
 In prophase II, the chromosomes CONDENSED / SPINDLE
FIBERS FORM
II. PROMETAPHASE II
 In prometaphase II, the NUCLEAR ENVELOPE
BREAKDOWN (if formed in telophase I) break down, and
the spindle organizes across the cell.
 Kinetochore microtubules from the opposite poles attach to
the kinetochores of each chromosome.
III. METAPHASE II
 In metaphase II, the movement of the kinetochore
microtubules aligns the chromosomes on the METAPHASE
PLATE
IV. ANAPHASE II
 During anaphase II, the centromeres separate, and the
now-daughter chromosomes are pulled to the opposite
poles of the spindle.
 One sister chromatid of each pair goes to one pole, and the
other goes to the opposite pole.
 The separated chromatids are now considered
chromosomes in their own right.
V. TELOPHASE II
 In the last stage, telophase II, the chromosomes begin
decondensing, a nuclear envelope forms around each set
of chromosomes, and cytokinesis takes place.
 After telophase II, the chromosomes continue
DECONDENSING eventually becoming invisible under the
light microscope.
 The end products of the two meiotic divisions are four
haploid cells (gametes in animals) from one original diploid
cell.
 Each of the four progeny cells has one chromosome from
each homologous pair of chromosomes.
 Because of crossing-over, these chromosomes are not
exact copies of the original chromosomes.
A COMPARISON OF MITOSIS AND MEIOSIS
 Basically, meiosis reduces the number of chromosome sets
from two (diploid) to one (haploid), whereas mitosis
conserves the number of chromosome sets.
 Mitosis - to make IDENTICAL CELLS
 Meiosis - to make different cells/ NOT IDENTICAL
o Therefore, meiosis produces cells that differ
genetically from their parent cell and from each
other, whereas mitosis produces daughter cells that
are genetically identical to their parent cell and to
each other.
THREE EVENTS UNIQUE TO MEIOSIS OCCUR
DURING MEIOSIS I:
 SYNAPSIS AND CROSSING OVER **only present in
meiosis I
o During prophase I, duplicated homologs pair up and
crossing over occurs.
o Synapsis and crossing over normally do not occur
during prophase of mitosis.
 HOMOLOGOUS PAIRS AT THE METAPHASE PLATE
o At metaphase I of meiosis, chromosomes are
positioned at the metaphase plate as pairs of
homologs, rather than individual chromosomes, as
in metaphase of mitosis.
o SEPARATION OF HOMOLOGOUS
o At anaphase I of meiosis, the duplicated
chromosomes of each homologous pair move
toward opposite poles, but the sister chromatids of
each duplicated chromosome remain attached.
o In anaphase of mitosis, by contrast, sister
chromatids separate.
o Meiosis I is called the REDUCTIONAL DIVISION
because it reduces the number of chromosome sets
from two (diploid) to one (haploid).
o During meiosis II (the EQUATIONAL DIVISION),
sister chromatids separate, producing haploid
daughter cells.
o The mechanism for separating sister chromatids is
virtually identical in meiosis II and mitosis.
ALVAREZ RMT’26 19
 Throughout Heaven and Earth, I alone am the Honored One

ORIGIN OF GENETIC VARIATION AMONG

OFFSPRING:
 Independent assortment of chromosome
 Crossing over
 Random fertilization

ALVAREZ RMT’26 20