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INTRODUCTION TO CELL BIOLOGY Cells Cells are very small (about 1-100 m). Cells act as functional basic unit of life. Robert Hook (1653-1703), an English Physician, firstly discovered dry cork cell by the earliest microscope that is a system of magnifying glass. He found that a regular shape of compartment that is cell wall on the dry cork. The foundation had been published in a local journal named Micrographia. Next, Mathias Schleiden and Theodor Schwan , a zoologist and botanist respectively ruled the cell theory or the cell doctrine (1839) which state: All organisms are composed of similar unit of organization called cells. Both previous achievement used as the foundation of the study of modern cell biology. The Principles of Cell Theory Cells are basic unit of life All living things made up of at least one cell Cells contain hereditary information (which closed to the cell division study) All cells are basically having the same chemical composition All energy flow in life occur within cells

General Characteristics of cells The shape: Highly depending on their functions Animal cells are likely irregular and regular for plants cells. Affected by the neighboring cells for positioning. Cell must be very small: Due to TSA/V, as size increase, the total surface area to volume decrease Due to nucleo-cytoplasmic ratio, nucleus can better control small cells. It is hardly to support bigger cell.

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Techniques In cell Study (1) Cell Fractionation (separating the cells components)

(i)

Homogenization This due to break the cell wall and cell membranes. The cells are prepared in - Isotonic solution To avoid extreme osmotic pressure to the cell - Chilled solution About 4C to deactivate enzymatic reaction or else, autolysis may occur and degenerating the cells organelles. For example, lysozymes from lysosomes. - Buffered solution Normally around pH 7.0 7.4 because cells are very sensitive to slight change of acidic or basic environment which may cause them degenerate. Usually these three solutions are integrated in detergent which provide easier way to decompose phospolipid bilayer at the plasma membranes.

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(ii)

Centrifugation The prepared homogenate inserted in test tubes which fix the centrifuges. These process described as follows, - Spin at high speed Homogenate will experience a very high centrifugal force which force them to the end of horizontal spinning test tube. - Homogenate separate due to density, speed and time Sample which has highest density will forced to accumulate faster, the less dense will force to pellet due to increasing in speed and time. Different organelles form pellet at different rate (differential centrifugation) - Collecting sample Suspended pellets then can be further purified by equilibrium centrifugation, which involves layering that solution on top of a density gradient ( a solution in which the concentration of the solute, usually sucrose, increases from top to bottom, forming a range of densities). When the density gradient centrifuged, pellet will segregate and form bands in the region that equal to their density. Cell fractions than collected by puncturing the tube bottom.

(2) Microscopic Techniques Anton Van Leeuwenhoek (1632-1723) firstly observed bacteria and protozoa through microscope. Microscopic parameters including (i) Magnification The higher the better to magnify objects. (ii) Contrast Ability to differentiate image with surrounding environment (iii) R , resolving power This is defined as the ability of a microscope to observe two neighboring objects separately. Resolving power is highly dependent to the light source.

RNA
Where NA is the numerical aperture. Therefore, shorter wavelength gives better resolution.

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(a) Light microscopes Also known as the phase contrast microscope.

(b) Electron microscopes Co-invented by Max-Knoll and Ernst Ruskol in 1931 Uses fine beam electrons Electrons are speeded up in a vacuum until their wavelengths are extremely short (about 0.004 nm) Beams are focused on sample Study dead specimen only The resolving power is about 0.2 nm. (i) Transmission Electron Microscope Electron transmitted through specimen To study internal structures The photograph produced called electron micrograph. Scanning electron microscope Electron transmitted through or on the surface of the specimen Specimen is coated with gold or platinum Produce 3D image and has a very large depth of field.

(ii)

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Prokaryotic cells and Eukaryotic cells Prokaryotic cells. Simply term as prokaryotes, it is single celled and the simplest. Mostly, they are from bacteria and archaebacteria. The cytoplasm/ protoplasm enclosed by plasma membranes. Obviously no cytoskeleton, the cell walls are made up of pepticloglycan to maintain the shape. As they have short lifespan, their metabolic rate and growth rate are super numerical. Membranous organelles are also absent. They have ribosomes to psynthesis their own protein. Special characteristic of this class including the mesosomes existence, which derived from their own plasma membrane that inveginate on purpose to produce energy. Furthermore, they possessed naked circular DNA in a region called nucleiod. Some of the cells have plasmid to store important genes, usually possesses by pathogenic bacteria. Other, they are gram negative and gram positive organism. Gram negative as cell wall thinner, gram stain doe not retain. Gram positive - thicker cell wall, retain the gram stain, usually purpled. Some prokaryotes have capsule which stick to their surfaces. Flagella also present to prokaryotes in need as movement aid in fluid environment. The hairy surfaces observed is actually pili which covered the whole organism and look like mini flagella. These prokaryotes usually reproduce through binary fission. Non Pathogenic bacteria

Photosynthetic - bacteriochlorophyll and photosynthetic enzymes Eg: purple bacteria - ingesting organic substances

Saprophytic Digestion occur outside the cell (release enzyme) Take digested food through plasma membrane without phagocytotic vesicles.

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(3) Eukaryotic cells Including animals and plants. They are all organism inclusive of prokaryotes. The cells bounded by plasma membranes. No cell wall for animals. The cells contain cytoskeleton such for supporting matrix of protein in animal. The three classes of organelles are: (i) Class 1 Endomembrane system. - In physical contact or pass tiny vesicle to one another - Relate by passing the vesicles - Example: RER, Golgi Apparatus (ii) Class 2 Mitochondrea and chloroplast have different type of membranes from others - Energy production agent - Contain own DNA and ribosomes thus, manufacture own protein - Believe to be the endosymbiotic bacteria (iii) Class 3 Ribosomes and nucleus Involve in gene expressions.

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General Comparison of Animal Cells and Plant Cells Animal cells Similarities Posses plasma membranes bounded the whole cells Have obviously dense nucleus as the central Varied cytoplasmic content functioning in different phase Animal cells Absent thus irregular shape at all Mitochondria Differences Cell wall Energy sources Plant cells Present and give regular shape Mainly chloroplast but some from mitochondria Starch droplet from glucose Regularly larger than animal Mostly disable especially multicellular none Large central vacuoles for matured, small also present Plant cells

Glycogen Regularly smaller than plant Some are motile especially unicellular Present for cell division Small if any

Form of stored food Size Mobility Centrioles Vacuole

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CELL STRUCTURES AND FUNCTIONS The Exterior structure (1) Cell Wall Tough and flexible but become rigid as the cell matured. It enclose the plasma membrane of plants. Cell wall are mainly found in - Plant (in form of cellulose) - Archaebacteria - fungi (in form of chitin) - algae Cell wall absent in animal and protozoa and mainly made up of cellulose. It protect the cell or plasma membranes from extreme environment and mechanical stress. (2) Plasma Membranes It enclose cytoplasm (protoplasm if for bacteria) to give protection to the cell. These membranes separating the interior part from the environment (interstitial fluid) and indirectly provide shape to the cell. As a semi permeable membrane, it allows the pass of certain substances at certain times. It adhesive ability made itself as the cell adhesive agent to stick to another cell or to a central organ. It provide ion channel and cell signaling which act as hormones detector. This means that certain cell with sensible to specific hormones only. These membrane is actually composed of phospolipid bilayer consist of polar heads and non polar tails. The History of Plasma Membrane Foundation Study (i) Danielli Darson model (1930s-1940s) They introduced that the plasma membrane consist of phospolipid bilayer which consist of polar heads that face the interstitial fluid and the non polar tails are hidden in between the heads. Plasma membrane is said to be semi permeable, thus they have perforated pores (about 1nm) at regular intervals It is static, stable and rigid structure.

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(ii)

Robertson (1959) He claimed that plasma membranes have no pores and posses a trilaminal structure Plasma membranes has the thickness about 7.5 -8.0 nm and he claimed that plasma membranes consist of two dense layer separated by a lighter layer The size is actually acceptable for one head and tail is about 3.5 to 4.0 nm

(iii)

Fluid Mosaic Model (early 70s) This model is purposed by Singer and Nicholson who use the freeze fracture techniques. The techniques are about freezing cell in solution and apply force to dissociate it component. They suggest that protein is not coating the plasma membrane and object Robertson by plasma membrane is actually dynamic fluid system which like jelly and keep flowing all the time. Globular protein also said to be present interspersed between the phospolipid. The membrane is selectively permeable and all exchange occurs is due to protein movement. (protein change it shaped, not energy come from substances) There are two types of proteins at the membranes, - Intrinsic protein which penetrating the layer including carrier and channel protein. - Extrinsic protein that acts as enzymes and hold the cytoskeleton.

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The Interior Structure (1) Cytoplasm Consist of cytosol in membrane bound organelles and each organelles then has their own cytoplasm. It formed like viscous gels which provide jelly like environment. Enclosed within the cell membranes, this is the site where most cellular activity occur such - Metabolism - Enzyme production - Glycolisis - Kreb cycle - End stage of ATP production Cytosol is the mix of cytoskeleton filaments, dissolve molecules in water. (2) Cytoskeleton This is the skeleton for cytoplasm which made up of filamentous protein network. Presents in animal and plant cells, this dynamic structure maintain the cell shape, protects the cell, and enable cellular motion by flagella, cilia or false foot projection. Cytoskeletons play important roles in intracellular transport and cell division

Microfilaments Each subunit is made up of actin Thinnest filament, flexible and strong Major portion of cytoskeleton Long fibres but thin with diameter about 7nm 2 chain twisted around one another Play role in muscle contraction Abundantly found in muscle cells and ameboid movement of certain cells Provide support
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Fillaments Made up of vimentin Most durable and stable The diameter varies around 8-12 nm Prevent excessive stretch of cells eg: keratin of skin cell (hardening) Shape and fix the organelles position

Microtubules Straight hollow rods Made up of and tubulin (proto filament) Form spontaneously around centrosomes Involved in many cellular process such transportation and give support during separation in cell division Part of ciliaand flagella of eukaryotic cells Unstable thus easily assemble and disassembled
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(3) Cilia Cilia are the projections of the cell, which is motile and the primary cilia is not motile. Cilia found on nearly every cell in human body which act seniority. All cilia have the same structures and same diameter around 0-2.5m, the length instead 2-20m. They are made up of microtubules covered by the plasma membranes. Functions - Movement - Sweep movement substances across the cell surfaces Arranged in rings, -primary: 9 pairs - Motile: 9 pairs surrounded by 2 central microtubules (9+2)

The basal body (centriole) anchor the cilium to the cell


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(4) Flagella - Projection from the cell - Made up of microtubules covered by plasma membranes - Longer than cilia (about 10- 200 nm) - For movement only - Basal body anchor flagellum to the cell - Arranged in rings. 9 pairs surround 2 ventral tubules (9+2)

(5) Nucleus Control centre of the cell. This largest organelle (about 5 m) is first to be discovered among the organelles. It is enclosed by a nuclear envelope : the double membrane perforated by nuclear pores ( about 100 nm) . The pores are actually regulates the entrances and exit of certain molecules. Nucleoplasm (not part of cytosol) is the semi fluid substance in the nucleus. The nucleus store DNA in fom of chromatin which become chromosomes during cell division and the most visible part of nucleus is nucleolus. Nucleus play a very advance role in synthesizing protein. Function (i) Maintain the genetic material by control any big changes to DNA (ii) Control the gene expression (forming protein) (iii) Mediate the replication of the DNA during the cell life cycle (iv) Provide sites for genetic transcriptions (v) Synthesis ribosomess and mRNA (vi) Allowing gene regulation (eg: lac operon control the lactose metabolism in certain bacteria)
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(6) Endoplasmic Reticulum Firstly observed under electron microscope by Keith Porter, Albert Claude, and Ernest F. Fullam in 1945. The term endoplasmic refer to within the cytoplasm and the other term reticulum means little nets This is the network of membranous tubules, vesicles, and flattened sacs called cisternae. The cisternal space or lumen is separated from cytosol by ER membrane The quantity of ER is depending on type and time of certain metabolic activity and the functions might be varies depending on the belonging cells. (a) Rough ER RER locate right after the nucleus and it membrane is continuous with the outer layer of the nuclear envelope. There are a lot of rough and flat cisternae, and significantly, a lot of ribosomes attached to the RER surface. The quantity of the ribosomes is not fized and depending to the protein amount as ribosomes is not a stable part- it is readily formed and breaks. The generalized protein journey in RER (i) protein enter the lumen (cisternal space) (ii) protein altered by enzyme (carbohydrate molecules normally added) (iii) secretory protein (iv) temporarily kept before being packed in vesicle and exported to Golgi Apparatus (v) proteins normally for export or membrane insertion (b) Smooth ER Found in a variety of cell types, thus, it function varies Obviously placed further from nucleus , no ribosomes attached, no protein production Enzymes anchored within smooth ER catalyse various process Example (i) (ii) (iii) In Gut Epithelium Enzyme of SER catalyse the synthesis of lipid Liver cells Carbohydrates metabolism (glucose glycogen) Testes and ovary Produce sex hormones and steroids
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(7) Ribosomes Two subunits of rRNA form ribosomes, solid spherical granules with diameter about 20-25 nm. An assembly of two ribosomal units synthesized in the nucleolus. Each subunit released through the nuclear pores. The ribosomes locate either bound to RER or in cytosol. For eukaryotic, larger made up of 60 subunits and smaller made up of 40 subunits only. Enzymes which synthesized by cytoplasmic ribosomes used within the cell while enzymes synthesized by RER-ribosomes exported outside. (8) Golgi Apparatus Named after Italian Physicist who discover it, Camellio Golgi This organelles made of a stack of flattened membranous sacs. In mammal, it consist about 5-6 but up to 20 in plant. The cisternal space space separated from cytosol.

The generalized protein journey in Golgi Apparatus:

Golgi Apparatus found abundantly in active cells and it is a part of endomembrane system A cis face is the receiving side located near the RER, vesicle from RER fuse with cis face and released the content. Protein modified during the transit from cis to trans pole by various enzymes producing varieties of oligosaccharides, a functional protein. Protein then marked and sorted into batches (capsuled) Then, the batches distributed to different destination in transport vesicles which bud fom trans face.(shipping side face plasma membranes) Protein move to the plasma membranes, leave the cell by vesicles, vesicle retain at the plasma membranes.
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(9) Lysosomes this is not a true organelles as the recycling centre of the cell. Autolysis recruited here as dead organelles digested here and whatever that can be reuse, lysis occur with the help of lysozymes. Pathogen and old organelles for example, are called autophagy. Mostly lysosomes in animal (up to hundreds per cell) but some in plants also. It digests macromolecules with enzymes sacs, most effectively at pH of 5. Lysosomes are produced internally at RER and just get similar to the enzyme production. RER put enzymes and membranes together, the Golgi Apparatus refined enzymes and release matured lysosomes. Digestive enzymes in lysosomes (named lysozymes ) are stored in compartment which divided by just thin membrane, thus it is said to be pseudoorganelles. (10) Mitochondria It is rod like structure and found abundantly in cells of active parts in human and found in animal for the energy source of respiration. The length is about 0.5-1.5 m and it width around 3-10 m. Mitochondria bounded by membrane; - Outer one give the shape - The mid space called intermembrane space - The inner one named cristae, which is folded in and partially divided into matrices. The cristae are tubular or villus like in plant while present in flattened sheet in animal. Cristae that facing the matrices covered with stalked particle. Mitochondria use glucose and fatty acids to produce ATP in Kreb Cycle. The matrix contains; CELL BIOLOGY

DNA Ribosomes Enzymes (the product of two aboves)


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(11) Chloroplast Large and flat disc, usually 5m by diameter and about 2-3 m thick. Possese its own DNA, ribosomes , and RNA. The double membrane separated by intermembranous space. The stroma inside contain; - DNA - Ribosomes - Photosynthetic enzymes (dependable in Kelvin cycle) 50 stacks of thylakoid form grana, 50 grana form internal membrane called lamellae. Photosynthetic pigments and electron transport chains (ETC) embedded with thylakoid membrane. Chloroplast usually contains; Oil droplets Starch granules

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TRANSPORT THROUGH A SEMI-PERMEABLE MEMBRANE These are three types of substances that generally pass through the semi permeable membranes (i) (ii) (iii) Small uncharged molecules including water molecules and gases. Water soluble substances Lipid soluble molecules

(1) Passive transport No energy is required and no force is applied as substances move from the higher concentration side to the lower one until they reach dynamic equilibrium. Molecules move in random motion produce kinetic energy itself and move across the plasma membranes. (a) Diffusion Cover all movements of all material, controlled by these three medium; - Temperature - Size of molecules - Concentration gradient Materials that undergo diffusion across the plasma membrane including; - Fat soluble molecules - Small molecules such as oxygen and carbon dioxide - Small polar but uncharged molecules like ethanol and water (b) Osmosis - only refer to water! Follow the concentration gradient, water move freely and collide each other at the level of water potential

Osmotic pressure, psi = 0 kPa


Osmotic pressure is the tendency for the solution to take up water separated by the semi permeable membrane Hypotonic solution: less solute, lower osmotic pressure Hypertonic solution: more solute higher osmotic pressure Isotonic solution: equal solute, equal osmotic pressure Shrinkage of cytoplasm when water moved out is called plasmolysis. In plant we called them plasmolysed but in animal cells, we refer it as crenation. Osmotic pressure is measured by osmometers. Hydrostatic pressure: the pressure of the cytoplasm pushing out against a membrane.
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(c) Facilitated diffusion No energy usage, follow the concentration gradient, carrier protein as the transport will change their shape when bind and release the solute. Normally they are very specific such only specific protein will adapt with specific substances. Substances undergo this diffusion are essential such; - Ions - Glucose - Amino acids (2) Active Transports. Against the concentration gradient is the significant thus, energy in form of ATP is badly required. Usually transport ion, glucose and amino acids. Involving protein pump, it occur mostly at - Kidney tubules - Small intestines - Nerve fibres - Plant roots

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(3) Endocytosis (a) Phagocytosis Take in solid substances / whole organism by invaginating the plasma membrane to form a vacuole. Usually run by white blood cell. (b) Pinocytosis Known as bulk-phase endocytosis which run the intake of liquid. The membrane infold and form pinocytic channel and pinocytic vesicles. (c) Receptor-mediated Endocytosis Recognise specific substances such cholera toxin, insulin and IgA (immunoglobin) Inner part of the membrane coated with clathrin which initiate the formation vesicles. Specific receptor will detect certain type of protein only. (4) Exocytosis This is the reverse process of endocytosis and used to export bulky material (usually protein produced by the cell). Vessicle will move to the membrane to remove its contain and usually endocytosis and exocytosis will occur at the same rate to maintain the total surface area, and size of the cell.

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