Cell

the basic structural and functional unit of living organism Some Vital Processes of Living Organisms Respiration Circulation Digestion & Absorption Excretion Reproduction Unicellular organisms: All vital processes occur in a single cell. e.g. Amoeba Multicellular organism: Specialized systems take over specific functions. Cells becomes specialized; organized into specific tissues, organs and hence systems. In humans, Gastrointestinal system digestion and absorption Cardiovascular systemtransport blood around the body; perfuse the tissues Respiratory system supply O and removes CO Urinary system excrete waste products; conserve volume and composition of body fluids Reproduction perpetuate species Control systems Nervous system (Quick component) Endocrine system (Slow component) Cells vary greatly in their size and shape. No cell can be referred to as the typical cell. But there are many features in common to all cells for maintenance of each cell's life

Cell Structure and Function

Chemical structure (chemical composition) The cell is composed of five basic substances:1. Water (70-80%) which acts as a medium for chemical reactions and transport of substances. 2. Electrolytes: Cat-ions: K+, Na+, Mg++ An-ions: HCO3, PO4, Cl 3. Proteins 4. Lipids and 5. Carbohydrates

Physical structure 1. Organelles are the living specialized structural parts of cytoplasm and nucleus. Organelles can be either membranous or non-membranous. nuclear membrane (or) Membranou Cytoplasmimembrane Nucleachromati NonNonribosomes cell envelope n membranous membranous c s r centrioles endoplasmic nucleolus reticulum microtubules, microfilaments Golgi apparatus or complex mitochondria lysosomes peroxisomes

Cytoplasmic membranous organelles: cell membrane, endoplasmic reticulum, mitochondria, Golgi apparatus or complex, lysosomes and peroxisomes Cytoplasmic non-membranous organelles: ribosomes, centrioles, microtubules and microfilaments Nuclear membranous organelle: nuclear membrane or envelope

Nuclear non-membranous organelles: chromatin and nucleolus

2. Inclusion bodies: non-living temporary components of cytoplasm viz. lipid globules, glycogen granules, secretory granules.

Cell membrane
the membrane surrounding the cell and the membrane of other organelles have many features in common also called the plasma membrane (or) unit membrane 2

Cell Physiology

Structure
it is a dynamic, rigid, semi-permeable structure ~7.5 to 10 nm thick composed of: proteins lipids phospholipids cholesterol Lipids phospholipids have the shape that resembles a clothespin they exists as a bimolecular layer with: the polar, hydrophilic (water soluble) heads (phosphate portion) facing: (1) the aqueous medium that bathes the exterior of the cell, and (2) the aqueous cytoplasm the non-polar, hydrophobic (water-insoluble) tails (fatty acid chains) facing each other in the middle. This arrangement imparts the fluidity to the lipid bilayer. Substances which are not lipid soluble have difficulty passing through the hydrophobic interior. This contributes towards selective permeability of the cell membrane. Globular Proteins are embedded in the fluid phospholipid bilayer matrix. Two types: 1. Integral proteins: They pass through the membrane and are the integral components of the membrane. Many of them have specific functions: e.g. transport proteins: contribute towards selective permeability of the cell membrane 2. Peripheral proteins: They stud the inside and the outside of the membrane. They are weakly bound the hydrophilic regions of specific integral proteins. Peripheral proteins held by glycosylphosphatidyl inositol anchors include: enzymes such as alkaline phosphatase various antigens a number of cell adhesion molecules

Cell Physiology

Functions of Cell Membrane

1. By forming a closed compartment around the cell, it allows cell individuality and maintains cell integrity. 2. Transport of substances into and out of the cell This is governed by the selective permeability of the cell membrane 2.1. Some proteins in the cell membrane function as "transport proteins" and they act as: (a) Channels: through which specific ions diffuse. Some channels are "continuously open". Some channels are "gated" i.e. they can be opened or closed by: (i) changes in the electrical potential (voltage) across the cell membrane (voltage-gated channels) (ii) binding of chemicals (ligands) such as hormones or neurotransmitters (ligand-gated or chemically-gated channels) (iii) other stimuli such as mechanical stretch. (mechanically-gated channels) (b) Carriers: they bind to the substances and translocate them from one side of the membrane to the other side. Carriers moving the substances along their concentration gradient facilitate diffusion of these substances. (c) Pumps: these are carriers that move the substances against their concentration gradient. They can hydrolyse energy phosphates (e.g. they have adenosine triphosphatase, ATPase, activity) and use the liberated energy for uphill transport of ions across the membrane. e.g. sodium-potassium ATPase pump

Cell Physiology

Transport proteins (carriers or pumps) may also be classified in a functional sense according to the number of substances moved and the direction of movement: A "uniport" transports only one substance (direction depends on the gradient). e.g. glucose transporter (GLUT) in muscle facilitates the glucose diffusion by moving glucose into the cells "Co-transporters" simultaneously transport more than one substance, and can be sub-categorized into two types: (1) a "symport" moves two or more different substances in the same direction (2) an "antiport" (countertransporter) moves two or more different substances in the opposite directions. e.g.i. Na-Ca antiport moves 2 Na into and one Ca out of cardiac muscle cells ii. Band 3 protein (anion exchanger) in RBCs and acidsecreting cells of the stomach moves one HCO and one Cl ion in opposite directions along their concentration gradients iii. Na-K pump moves Na and K in opposite directions against their concentration gradient. 2.2. The cell membrane exhibits endocytosis (pinocytosis=cell drinking, phagocytosis=cell eating) and exocytosis (emeiocytosis=cell vomiting), transporting particulate matter and proteins into or out of the cells. 3. Other proteins in the cell membrane function as (a) receptors, which provide binding sites for many molecules e.g. hormones and drugs (b) enzymes, catalyzing reactions at the surfaces of the membrane (c) antigens, carrying immunological identification marks e.g. blood group antigens (d) cell adhesion molecules (CAMs) These molecules attach cells to each other and to the basement membrane, forming intercellular connections which give strength and stability to tissues. Many CAMs pass through the cell membrane and are anchored to the cytoskeleton. Others bind to large molecules in the extracellular matrix. They play important role in: i. development and formation of nervous system and other tissues ii. holding tissues together for structural support (by CAMs called cadherins) iii. inflammation iv. wound healing and scar formation v. spread of tumors vi. transmission of signals into and out of the cells

Cell Physiology

INTERCELLULAR CONNECTIONS Two types 1. The junctions that the cells together giving strength and stability to tissues a. Tight junctions (zonula occludens) made up of protein ridges - half from one cell and half from adjacent cellstrongly held together found in apical margins of epithelial cells in the intestinal mucosa, renal tubules and the choroid plexus form a barrier to the movement of ions and other solutes from one side of the epithelium to the other but permit passage of some ions and solutes between cells (Paracellular pathway) although the degree of leakiness varies from site to site b. Desmosomes spot-like patches characterized by apposed thickening of the membranes of two adjacent cells c. Hemi-desmosomes half desmosomes that attach cells to an underlying basal lamina d. Zonular adherans continuous structures on the basal side of the zonular occludens major site of attachment for intracellular myofilaments 2. The junctions that permit transfer of ions and molecules from one cell to another (Gap junctions) forms a cytoplasmic "tunnel" for diffusion of small molecules (< 1000 Da) between two neighboring cells consists of hexagonal arrays of protein units called connexons in the membrane of each cell, lining up with each other forming a single channel bridging the membranes of two cells permit substances (ions, sugars, amino-acids) to pass between cells without entering the ECF, enabling rapid propagation of electrical activity from cell to cell and the exchange of chemical substances found in cardiac and smooth muscles (forms the physical basis of "functional syncytium")

ENDOPLASMIC RETICULUM
it is a complex series of tubules extending throughout the cytoplasm. the walls of the tubules are made up of unit membrane and is continuous with the outer nuclear membrane, golgi apparatus and cell membrane. serves as "intracellular circulatory system"

Cell Physiology

Two types of ER
1. Granular (or) Rough-surfaced Endoplasmic Reticulum granules called ribosomes are attached to the cytoplasmic side of the membrane the attached ribosomes are the site of synthesis of proteins such as hormones that are secreted by the cell and proteins that are segregated in lysosomes the polypeptide chains that form these proteins are extruded into the ER 2. Agranular (or) Smooth-surfaced Endoplasmic Reticulum lacks granules or ribosomes Functions: a. Metabolic function: as site of i. steroid synthesis (in steroid secreting cells) ii. detoxification (e.g. liver cells) iii. glycoprotein synthesis b. as Sarcoplasmic Reticulum, plays important role in initiating muscle contraction and relaxation of cardiac muscle and skeletal muscle

Cell Physiology

Golgi apparatus (Golgi complex or Golgi bodies)

a collection of membrane enclosed sacs (cisterns) that are stacked like dinner plates continuous with the ER and is usually located near the nucleus particularly prominent in actively secreting cells polarized (cis- = same: on the side of nucleus; trans-= opposite: on opposite site) Membranous vesicles containing newly synthesized proteins bud off from the granular endoplasmic reticulum and fuse with the Golgi apparatus on the cis- side. Then, the proteins are passed on to the cistern on the trans side. During transit, proteins are provided the code for their final destination (e.g. proteins for lysosomes, proteins to be secreted etc.) and the modified proteins are repackaged into vesicles.

Function: package the proteins with membranes and modifies their carbohydrate moieties, preparing them for their final destination

Mitochondria
(singular: mitochondrion) Mitochondria are present in almost all cells The more active the cell, the greater the number of mitochondria.

Structure
Sausage-shaped structure made up of double layer of unit membrane The outer membrane smooth contains enzymes of biologic oxidation The inner membrane folded into projections called cristae mitochondriales The space between the membranes: intercristal space The space surrounding the cristae: matrix contains enzymes of Krebs citric acid cycle On the cristae: repeating units contains enzymes of respiratory chain & ATP synthase These enzymes and the enzymes of Krebs Citric Acid Cycle convert the products of carbohydrate, fat and protein metabolism (oxidation) to CO and water. Electrons are transferred along the respiratory-enzyme chain resulting in the synthesis of the high energy phosphate compound, adenosine triphosphate (ATP). The process is known as oxidative phosphorylation. During the process, protons are pumped from the matrix Cell Physiology 8

into the intercristal space, and the protons diffusing back by its own gradient drive ATP synthase for new ATP synthesis. Functions: 1. Mitochondria are power generating units of the cell because they produce ATP (essential for aerobic energy production) 2. Mitochondria contain DNA which represents a second genetic system in the cell. The DNA has cyclical arrangement (normal nuclear DNA: double helical arrangement) and inherited only from mother. DNA repair system is poorly developed. higher mutation rate leading to diseases such as myopathies and are associated with aging (senescence) and diabetes mellitus. 3. Mitochondria can serve as an intracellular storage site for Ca.

Lysosomes
large irregular structures surrounded by unit membrane each contains a variety of enzymes that can cause destruction of most cellular components ribonucleases deoxyribonucleases phosphatases glycosidases arylsulfatases collagenases cathepsins the interior is more acidic than the cytoplasm (~pH 4.5)

Functions: function as the digestive system of the cell 1. Defence and scavenger function by digesting the phagocytosed foreign particles such as bacteria. Some products of digestion are absorbed through the wall of the vacuole and some are excreted by exocytosis 2. Engulfment and removal of the worn-out components of the cell 3. Autolysis of dead cells 4. help in cellular differentiation and regression of tissues e.g. involution of uterus

Cell Physiology

Lysosomal dysfunction In vitamin A intoxication and certain other conditions, lysosomal membranes break down with the release of enzymes into the cytoplasm thereby destroying cellular components. In gout, phagocytosis of uric acid crystals by neutrophils triggers the release of lysosomal enzymes that contribute to inflammation of the joints. Congenital absence of any one of the lysosomal enzymes leads to engorgement of the lysosome with the material the enzyme normally digests and eventual disruption of the cell (lysosomal storage diseases)

Peroxisomes
about 0.5 m in diameter surrounded by unit membrane contain various oxidases (which catalyze reactions generating HO )and catalase (which converts HO to O and HO more common in liver and kidneys may be involved in gluconeogenesis destroy a number of compounds and detoxify fatty acids metabolize ethanol to acetaldehyde

Cytoplasmic non-membranous organelles Ribosomes

are granules with two subunits: 40s and 60s the site of protein synthesis contains RNA (65%) and protein (35%) Ribosomes attached to the ER synthesize proteins to be secreted by the cell. (e.g. hormones) Free ribosomes in the cytoplasm synthesize proteins to be utilized within the cell. (e.g. haemoglobin)

Centrioles
a pair of short cylindrical structures located between the golgi complex and nucleus arranged at right angles to each other the wall is made up of microtubules

Function: concerned with movement of chromosomes during cell division forming poles of the mitotic spindle Cell Physiology 10

Microtubules and microfilaments

Microtubules are long, hollow structures about 15 nm in diameter. Microfilaments are long, solid structures about 4-6 nm in diameter. Functions: Microtubules provide tracks along which secretory granules are moved from one part of the cell to another. Protein molecules with ATPase activity that move various substances along microtubules are called molecular motors (kinesin and dynein) Microtubules form mitotic spindles which move chromosomes during cell division, mitosis. Microtubules, microfilaments and proteins that tie them together to form the skeleton of the cell. The cytoskeleton not only maintains shape but also permit to change shape and move. Microfilaments are made up of actin. All cells contain actin and a variety of actin-binding proteins, including myosin (an actin-based motor). They play a role in: muscle contraction movement of microvilli in the intestinal mucosa movement of the cell and clot retraction Intermediate filaments connect the nuclear membrane to the cell membrane, and also help the cell to resist external pressure. Cells rupture more easily and skin blisters are common when these filaments are absent or abnormal.

Nucleus
A nucleus, the central information centre, is present in all animal cells that divide. It consists of: 1. Nuclear membranous organelle: Nuclear membrane or envelope 2. Nuclear non-membranous organelles: Chromatin and Nucleolus 1. Nuclear membrane It is a double unit membrane, 25~40nm thick, enclosing a space called the peri-nuclear cistern. The pores in the nuclear membrane are closed by a thin membrane. The membrane permits passage of molecules as large as RNA. 2. Nuclear non-membranous organelles: a. Chromatin Chromatin are granules of dark, densely stained particles which condense to form chromosomes during cell division Cell Physiology 11

Chromosomes are made up of DNA and a basic protein called histone. The chromosomes carry the genetic message, the complete blueprint for all the inheritable characteristics of the cell. The functional unit of DNA is a gene which contains the genetic information required to form one polypeptide or protein molecule (e.g. all the enzymes which control the metabolism of the cell) Human have 46 chromosomes, 22 pairs of somatic chromosomes (autosomes) and 2 sex chromosomes (X and Y in males; two X in females). Total number of Autosome Sex chromosome Ploidy chromosome s Somatic 46 44 2 Diploid cell (23 pairs) (22 pairs) (1 pair) (XY in male, XX in female) Germ cell 23 22 1 Haploid (not in pairs) ploidy = the multiple of the basic number of chromosomes in a cell The complete genetic information is represented by the haploid number of chromosomes (~3x10 base pairs in 23 separate molecules) b. Nucleolus a patch-work of granules with no limiting membrane. The granules are rich in RNA. Functions: the site of synthesis of ribosomal RNA (rRNA) temporary storage of messenger RNA (mRNA)

Deoxyribonucleic acid (DNA) a double helix formed by 2 chains of polydeoxyribonucleotides held together by hydrogen bonds made up of phosphoric acid pentose sugar nitrogenous base Purines Adenine & Guanine Pyrimidines Cytocine & Thymine Adenine always pairs with Thymine (A : T) Guanine always pairs with Cytosine (G : C) Cell Physiology 12

The total number of purines = the total number of pyrimidines The two chains are: complementary (with regards to base pairing) anti-parallel (oriented in opposite directions)

Each chain is made up of deoxyribonucleotide units linked by 3',5'phosphodiester bonds. The genetic message is coded by the sequence of the bases in the deoxyribonucleoide chains. The message is transferred to the sites of protein synthesis by RNA. The text of the message is the order in which the amino acids are lined up in the protein synthesized.

Function: carrier of genetic information serves as template for replication (formation of new DNA during cell division) serves as template for formation of RNA responsible for maintenance of the species (passed from one generation to the next) highly preserved through generations

Ribonucleic acid (RNA) A single strand of ribonucleotides, made up of: phosphoric acid pentose (ribose) sugar nitrogenous base purines: Adenine, Guanine pyrimidines: Thymine, Uracil mainly present in the cytoplasm 1. Ribosomal RNA (rRNA) found in ribosomes (Ribosomes contains 65% RNA and 35% proteins) involved in the translation of genetic message 2. Transfer RNA (tRNA) (soluble or acceptor RNA) at least 20 tRNA molecules in every cell (one for each of the 20 amino acids) tRNA is arranged in clover leaf structure tRNA molecules carry activated specific amino acids to the sites of protein synthesis (=ribosomes)

Cell Physiology

13

serves as adaptors for translation of the information in mRNA into specific sequence of amino acids

3. Messenger RNA (mRNA) mRNA carries sequence of nucleotides complementary to the sense strand of DNA serves as messenger conveying genetic information from the nucleus to the ribosomes (the site of protein synthesis) where it in turn serves as template on which a specific sequence of amino acid is polymerized to form a specific protein.

Protein Synthesis
The Genetic Code
Genetic information lies in the sequence of nucleotides in DNA The information is transcribed from DNA to mRNA. Each information code word exists for each amino acid. The code word is termed codon and consists of a triplet of nucleotides (either A, C, G or U). there are 64 (= 4) possible codons. 61 codes for amino acids. 3 serves as terminating signals or nonsense codons i.e. translation stops when it encounters stop codons (UAA, UAG, UGA) More than one codon codes for a given amino acids. e.g. Both UUU and UUC stands for phenylalanine; six codons codes for serine.

The process of protein synthesis 4 main stages: 1. Transcription 2. Post-transcriptional modification 3. Translation 4. Post-translational modification

Cell Physiology

14

1. Transcription In transcription, the code (information) on the sense strand of DNA (template) is transcribed into a sequence of nucleotides (mRNA) by the action of DNA-dependent RNA polymerase. The synthesized mRNA is also called pre-mRNA or hnRNA (heterogeneous nuclear RNA). 2. Post-transcriptional modification The newly synthesized mRNA is processed before it is released into the cytoplasm. The mRNA thus formed is called the definitive mRNA. 3. Translation This step takes place in the ribosomes. The information carried by the mRNA is translated from the linear sequence of codons (nucleotide triplets) into a linear sequence of amino acids (peptides or proteins). tRNA, each loaded with its specific amino acid, serves as adaptors for assembling of amino acids. 4. Post-translational modification The newly formed polypeptide chain is modified by one or more chemical reactions into a final protein to achieve its functionality. e.g. prepro-insulin pro-insulin insulin

Cell Physiology

15

Transport Across Cell Membranes

1. Passive Transport Mechanisms a. Diffusion i. Simple or free diffusion ii.Facilitated or carrier-mediated diffusion iii.Non-ionic diffusion b. Osmosis 2. Active Transport Mechanisms a. Primary active transport b. Secondary active transport c. Endocytosis d. Exocytosis Transport across epithelia 1. Filtration 2. Transcytosis (Vesicular Transport)

PASSIVE TRANSPORT MECHANISMS

In passive transport mechanisms, the movement of a substance (or) substances: occurs spontaneously does not depend on supply of metabolic energy and is downhill along the gradient(s): concentration or chemical gradient (from an area of higher concentration to an area of lower concentration) electrical gradient (from an area with same electrical charge to an area with opposite charge) pressure gradient (from an area with higher hydrostatic pressure to an area with lower hydrostatic pressure) osmotic gradient (from an area with lower osmotic pressure to an area with higher osmotic pressure)

(a) Diffusion Diffusion occurs due to random thermal motion of molecules in fluid state (liquid or gas state). Diffusion is the continuous random movement of molecules and ion from a region of higher concentration to a region of lower concentration along gradients (concentration gradient, electrical gradient, pressure gradient or osmotic gradient). Cell Physiology 16

"flux" denotes the transfer of substances across a unit area in a given time "influx" = flux into the cell "efflux" = flux out of the cell

Cell Physiology

17

The magnitude of diffusion across the cell membrane is governed by (1) the properties of the substance (i) size (ii) electrical charge (iii) lipid solubility (2) the barrier itself (i) the permeability (ii) the thickness (iii) the cross-sectional area (3) the forces across the cell membrane (concentration or chemical gradient, electrical gradient) Fick's Law of Diffusion The net rate of diffusion (or) flux (J) across the membrane is: directly proportionate to: the area available for diffusion (A) the diffusion coefficient of the membrane for the diffusing substance (D) the concentration gradient (C) and inversely proportinal to: the thickness of the membrane (or the length of diffusion path, x) that is, J=D A Cx e.g. The net rate of diffusion of oxygen in the inspired air across the pulmonary membrane into the blood (J) will depend on: 1. the partial pressure gradient (the difference in partial pressure) between the air and blood (P) 2. the diffusion capacity of the pulmonary membrane (lungs) to Oxygen (D) 3. the area of the pulmonary membrane (A) 4. the thickness of the pulmonary membrane (x) Any decrease in area available for diffusion (due to destruction of the lungs) or any increase in the thickness of the pulmonary membrane due to disease process will greatly reduce the oxygenation of blood. Selective Permeability of Cell Membrane Cell membranes are impermeable to proteins and other organic substances such as phosphates. Such ions are called non-duffusible ions. The presence of non-diffusible ions on one side of a semipermeable membrane causes unequal distribution of of diffusible ions across the semipermeable membrane. Cell Physiology 18

Diffusible substances Diffusible ions can cross the cell membrane easily. These include: Lipid soluble molecules (e.g. steroids) Non-polar molecules (e.g. O, N) small uncharged polar molecules (e.g. CO)

Non-diffusible substances Non-diffusible substances cannot pass through the cell membrane easily, and thus require transport proteins to pass through. Proteins Small, charged substances ( e.g. ions: H, Na, K, Mg, Ca, Cl) Large, uncharged molecules (e.g. glucose) Polar molecules of medium to large size (e.g. organic acids s/a RCOOH)

(i) Simple diffusion Diffusion of substances across the lipid bilayer or diffusion of ions through the ion channels is called simple diffusion. e.g. Steroid hormones dissolve in the lipid bilayer of the membrane and cross with ease e.g. K moves out of the cell membrane via K channel (ii) Facilitated diffusion Diffusion of large uncharged molecules across the lipid bilayer becomes greatly facilitated when they are moved across the cell membrane along their chemical or electrical gradients by transport proteins called "carriers". When the carrier proteins bind to the substance to be transported, their configuration changes so that the bound substance is moved from one side of the membrane to the other. e.g. Transport of glucose by the glucose transporter (GLUT), which moves the glucose down its concentration gradient from the ECF into the cytoplasm of the cell (iii) Nonionic diffusion Non-ionic diffusion is the diffusion of some weak acids and bases in the undissociated (non-ionic) form. In the ionic form, diffusion becomes difficult. e.g. NH3 can readily diffuse across the renal tubular cell membrane but NH ion cannot. (b) Osmosis DEFINITION: If 2 solutions of equal volumes but of unequal strengths are separated by a membrane permeable to the "solvent" (e.g. "water" in most cases) but not to the "solute", the solvent (water) will move across the membrane from the side with lower concentration of the solute (i.e. with higher concentration of water) to Cell Physiology 19

the other side. This type of solvent movement is known as OSMOSIS. Essentially, it is the diffusion of water along its concentration gradient. The pressure necessary to apply on the more concentrated solution to prevent solvent movement is defined as the "effective osmotic pressure" of the solution. Osmotic pressure depends on the number of particles in a solution rather than the type or chemical nature of particles. The concentration of osmotically active particles is expressed in osmoles. Osmosis is important because it is the major mechanism by which water moves across biological membranes.

Filtration Filtration is the process by which fluid is forced through a membrane or barrier (a capillary endothelial wall) due to the difference in hydrostatic pressure on the two sides. The amount of fluid filtered is proportionate to the difference in pressure (hydrostatic pressure gradient), the surface area and the permeability of the membrane. Molecules smaller than the pores of the membrane pass through along with the fluid; larger molecules are retained. Filtration through capillary wall is termed ultrafiltration since not only particulate matter like blood cells but also colloids like proteins are retained. Solvent drag When the filtering membrane is very permeable, the amount of fluid flowing in one direction becomes very large (bulk flow), and the solvent tends to drag along some molecules of solute along with it. This is called "solvent drag". e.g. This is seen in glomerular filtration where the permeability of the glomerular capillaries is very high.

ACTIVE TRANSPORT MECHANISMS

Active transport requires metabolic energy and the movement is usually uphill i.e. against concentration or electrical gradients.

(a) Primary active transport

Active transport is carried out by "protein pumps" in the cell membranes and the energy is supplied by adenosine triphosphate (ATP) generated by the metabolism of cells ("metabolic energy"). Examples: 1. The sodium-potassium pump (Na+-K+ ATPase)

Cell Physiology

20

It has Na+-K+ activated ATPase which catalyzes the hydrolysis of ATP to ADP and uses the liberated energy to extrude 3Na+ from the cell and take 2K+into it (coupling ratio = 3:2) for each molecule of ATP hydrolyzed. It produces net movement of a positive charge out of the cell for every cycle, and creates an electrical charge difference across the cell membrane. Therefore the pump is electrogenic. The Na+-K+ pump is found in almost all cells. Active transport of Na+ and K+ is one of the major energy-using processes in the body accounting for a large part of the basal metabolism. On average: 24% of energy utilized by the cell (~70% in neurons)

Structure: a heterodimer (one subunit + one subunit) subunit have intracellular binding sites for Na and ATP and extracellular binding sites for K Mechanism: When 3 Na bind to their binding sites on subunit accessible only from inside the cell, one ATP molecule also binds and converted into ADP, with a Phosphate transferred to subunit (phosphorylation). This causes change in configuration of the protein, extruding 3 Na into the ECF. K then binds from ECF, dephosphorylating the subunit, which returns to previous configuration, releasing K into the ICF. Regulation: Intracellular [Na]: the greater the [Na]ICF, the more the Na is pumped out (never saturate) Second messengers: cAMP, Diacyl glycerol (DAG) Thyroid hormones: pump activity by stimulating new formation of Na, K ATPase Aldosterone increases the no. of pumps Dopamine inhibits the pump (by phosphorylating it) Insulin increases the pump activity. It can be inhibited by Ouabain and Digitalis in the treatment of heart failure

Cell Physiology

21

2. H+-K+ ATPase: extrudes H+ from the cells in exchange for K+ 3. Ca2+ ATPase: pumps Ca2+ out of cytoplasm into the endoplasmic reticulum in skeletal and cardiac muscle cells. This leads to muscle relaxation. 4. V-ATPase: pumps H+ (protons) out of cytoplasm into organelles (lysosomes, parts of Golgi apparatus) 5. F-ATPase (ATP synthase): pumps H from mitochondrial matrix into the intercristal space, setting up the proton gradient essential for ATP synthesis

Cell Physiology

22

(b) Secondary active transport

Some carriers transport one or more molecules against their respective electrochemical gradient (inward or outward; active / uphill) by using the energy in the electrochemical gradient of the other molecule or molecules to drive this transport (outward; passive / downhill). The electrochemical gradient of the latter molecule (Na in most cases) is maintained by ATP using pumps that transport it out of the cell. 1. Sodium-dependent glucose transport: The cell membranes (apical or luminal) of intestinal and renal tubular cells contain a co-transport protein (symport) that transport glucose into the cells only if Na binds to it. The movement of sodium is always downhill, while the net movement of glucose is uphill, moving from lower to higher concentration. Na gradient is maintained by the active transport of Na out of the cell by Na, K ATPase pump.
SGLT 2 can generate an approximate 100-fold glucose gradient SGLT1 can generate a nearly 10,000-fold glucose gradient assuming an intracellular [glucose] of 2 mmol/L, SGLT1 could remove virtually all glucose from either the lumen of the small intestine or the lumen of the proximal tubule (i.e., luminal [glucose] of 0.0002mmol/L).

2. Sodium-dependent Ca2+ transport The cardiac muscle cell membranes contains an exchange protein (antiport) which extrude one Ca ion against the electrochemical gradient for three Na taken into the cell. The rate of this exchange is proportional to the concentration gradient of Na across the cell membrane which in turn depend on the activity of the Na, K ATPase pump.

Cell Physiology

23

(c) Endocytosis
Endocytosis is the process by which proteins and large molecules enter the cell without disruption of the cell membrane. The cell membrane folds inwards and pinches off to form a tiny sphere of membrane (called the vesicle) which encloses the ECF and the substances that are being transported. With endocytosis, there is loss of cell membrane enveloping the cell. There are 2 types:1. Phagocytosis (cell eating): It is the process by which large particulate matter (i.e. not in solution in the body fluids) such as bacteria, dead tissue, or other bits of material visible under the microscope are transported into the cell. e.g. phagocytosis of bacteria by macrophages Mechanism: the particle makes contact with the cell membrane cell membrane invaginates at the area of contact the cell extends pseudopodia around the particle (engulfment) pseudopodia fuses to enclose the particle within a vacuole (phagocytic vacuole or phagosome) phagosome fuses with lysosome (phagolysosome or digestive vacuole) enzymatic digestion occurs followed by absorption into the cytoplasm undigestible materials are extruded out of the cell by exocytosis (cell defecation)

1. Pinocytosis (cell drinking): It is the process by which "large molecules in solution" (e.g. proteins) are transported into the cell. Mechanism: the material (in solution) makes contact with the cell membrane invagination occurs at the area of contact pinching off of the invagination formation of pinocytic vacuole which may: pass through the cell unaltered (in case of capillaries or intestinal wall) Cell Physiology 24

or may combine with intracellular organelles such as lysosomes to form digestive vacuole Two types of Endocytosis: 1. Constitutive endocytosis: It is a continuous process that is not induced. The uptake is simply proportionate to the concentration of the substance in the surrounding ECF e.g. the uptake of plasma proteins by endothelial cells of capillaries 2. Non-constitutive or receptor mediated endocytosis It is a receptor mediated selective process for internalization of macromolecules such as protein hormones (insulin, growth factors), LDL, toxins occurs at cell membrane indentations where the protein , clathrin, accumulates triggered by various ligands binding to their receptors on the cell surface

(d) Exocytosis (Reverse endocytosis or cell vomiting)

Exocytosis is the extrusion process by which cellular secretions (proteins and large molecules) are liberated to the exterior. e.g. secretion of protein hormones and enzymes. Proteins that are secreted by cells move from the endoplasmic reticulum to the Golgi apparatus where they are packed into secretory granules or vesicles. The granules and vesicles move to the cell membrane. Their membrane then fuses with the cell membrane and the area of fusion breaks down. This leaves the contents of the granules or vesicles outside the cell and the cell membrane intact. Exocytosis adds to the total amount of membrane enveloping the cell. The process requires Ca2+ and energy, and "docking proteins" that dock the secretory granules or vesicles to the cell membrane. * Docking proteins: v-SNARE protein, t-SNARE protein Proteins may be exocytosed: 1. with little or no prior processing or storage (Constitutive pathway) 2. after processing in the secretory granules (Non-constitutive pathway) e.g. conversion of prohormones to mature hormones

Cell Physiology

25

Vesicular transport (Transcytosis or Cytopempsis) Small amounts of protein are transported out of capillaries across endothelial cells by endocytosis followed by exocytosis on the interstitial side of the cells. The transport system makes use of coated vesicles and is called vesicular transport (transcytosis or cytopempsis).

INTERCELLULAR COMMUNICATION
Cells communicate with each other via "chemical messengers". Within a given tissue, some messengers move from cell to cell via "gap junctions" without entering the ECF. Cells are also affected by chemical messengers secreted into the ECF. These chemical messengers bind to protein receptors on the surface of the cell (or) in the cytoplasm or the nucleus of the cell, triggering a sequence of intracellular changes that produce their physiologic effects. There are 3 general types of intercellular communication mediated by messengers in the ECF: (1) Neural Communication, in which neurotransmitters are released at synaptic junctions from nerve cells and act across a narrow synaptic cleft on a post-synaptic cell. (2) Endocrine Communication, in which hormones reach cells via the circulating blood: (3) Paracrine and Autocrine communication, in which the products of cells diffuse in the ECF to affect neighboring cells that may be some distance away (paracrine communication) or bind to receptors on the cell that secreted them (autocrine communication). Some growth factors are attached extracellularly to the transmembrane proteins of some cells. Such a factor anchored to a cell can bind to its receptor on another cell, linking the two. This is called juxtacrine communication and may be important in growth in tissues. Table 1 Intercellular communications 26

Cell Physiology

Cell Physiology

27

28