CELL AND TISSUE BLOCK PHASE 1 2008/2009 Transport across cell membrane

Objectives:
Outline the basic concepts of transport across cell membrane with regard to 1. osmosis 2. diffusion 3. facilitated diffusion 4. active transport (symport, antiport) 5. ionophores 6. exocytosis 7. endocytosis

Dr Adibah Alawiah bt Hj Abd Razak MD (USM), MSC (Physiology, RCSI) Ireland

Q1 : Why should I study about Transport across cell membrane?

Q2 : Answer :

Transport across cell membranes can be divided into two main categories : Membrane transport systems

Passive or spontaneous transport Simple diffusion Facilitated diffusion

Active or energy requiring transport Primary active transport Secondary active transport

Carrier independant

Channel/Carrier dependant

Passive transport (1) Simple diffusion

Diffusion in a mixture occurs because of the random movement that all particles exhibit due to their thermal energies This random thermal movement is also known as Brownian motion Simple diffusion = net movement of a substance from an area of high concentration to an area of low concentration. The magnitude and direction of the net movement of particles are determined by concentration gradient

Continue Passive transport

(1) Simple diffusion

Flux is the general term used to describe the rate of movement of solute molecules It is expressed in terms of amount of solute moving per unit time

Continue Passive transport (1) Simple diffusion

Lipid solubility The rate at which a solute diffuses through the membrane depends on two factors 1. 2. Its ability to dissolve in lipids Its molecular size

Continue Passive transport (1) Simple diffusion

Osmosis
Tendency of water to move from a dilute solution to a more concentrated solution Osmosis can be viewed as the passive diffusion of water or more generally any solvent down its concentration gradient across a selectively permeable membrane Osmotic pressure is determined by the number of osmotically active particles in solution and is effectively independent of particle size 1M solution of glucose, 0.5 M solution of NaCl, 0.333M solution of CaCl2 have approximately the same osmotic pressure

In general a non-polar and therefore lipid soluble solute (e.g. urea, CO2, N2, O2) will diffuse across a cell membrane much more rapidly than a polar (water soluble) solute of the same size

Osmosis = diffusion of water across a semi permeable membrane, from an area of low solute concentration to an area of high solute concentration

Continue Passive transport (2) Facilitated or carrier-mediated diffusion

Simple diffusion occurs via the lipid bilayer

In facilitated diffusion the transport of a substance is via membrane channels or transporters/carriers (pumps are excluded because they mediate active transport). Channels are usually uniporters whereas carriers can be uniporters, symporters or antiporters

Continue Passive transport (2) Facilitated or carrier-mediated diffusion

Continue Passive transport (2) Facilitated or carrier-mediated diffusion

Examples of facilitated diffusion through transporters and channels are: 1. 2. Transport of urea using urea transporters Transport of water through water channels

Urea is transported via transporters. Four different urea transporter isoforms have been identifed UT-A1 expressed in the apical membrane of principle cells of the terminal collecting duct. Sensitive to ADH expressed in the apical membrane of thin descending limb of the short loops of Henle and involved in urea recycling. expressed in erythrocytes, inner and outer medulla of the kidney, descending vasa recta, testis, brain and spleen this is identical to the N-terminal half of UT1 and expresse in the kidney

Cell membranes in the human body have a permeability to water ranging from zero in the thick ascending limb of the loop of Henle to flow velocities of 500 msec-1 in the RBC and renal proximal tubules Movement or diffusion of water occurs passively through the lipid bilayer or through specific pores or channels Water pores in the cell membranes are referred to Aquaporins AQP1 proximal tubule, RBC, capillary endothelia, outer medullary descending vasa recta apical membrane of principal cells in the corticol duct (vasopressin sensitive) basolateral membrane CD epithelial cells, airway, colon, (vasopressin sensitive) basolateral membrane of collecting duct, airway surface epithelia, CNS (BBB), inner ear, retina Glandular epithelia (salivary gland) , corneal epithelium, alveolar epithelium, GI tract glomerular podocytes, prox. tubule, collecting duct

UT-A2 -

AQP2 AQP3

UT-A3 -

AQP4

UT-A4 -

AQP5 AQP6

AQP7 AQP8 AQP9

proximal tubule, heart, adipose tissue, testes Liver pancreas, intestine, salivary gland, testis, heart Liver, white blood cells, testes, brain small intestine Kidney, liver Pancreatic acinar cells

Aquaporins and diseases:

Mutations in the aquaporin-2 gene cause hereditary nephrogenic Diabetes Insipidus in humans A small number of people have severe or total deficiency in aquaporin-1. they are generally healthy, but exhibit a defect in the ability to concentrate solutes in the urine and to conserve water when deprived of drinking water. autoimmune reactions against aquaporin 4 produce Devics disease

AQP10 AQP11 AQP12 -

AQP1, 2, 4, 5, and 8 are primarily water selective , whereas AQP3, 7, 9, 10 also transport glycerol and therefore called aquaglyceroporins Aquaporins are small hydrophobic, integral proteins ( 30kDA/monomer) In most cases they reside constitutively at the plasma membrane, except for AQP2 and AQP3 in the kidney, which undergo vasopressin-regulated exo-endocytosis similar to insulin regulated GLUT-4

** Peter Agre (Johns Hopkins School of Medicine) shared 2003 Nobel Prize in Chemistry for his work on aquaporins. Nice to know things!

The facilitated glucose transporter family Two families of glucose transporters have been identified 1. Na+-dependant glucose transporters (SGLT) 2. Facilitated-diffusion glucose transporters (GLUT) GLUT 1-5 and & 7 have been identified : GLUT 1high affinity glucose transporter Expressed in RBC and BBB, proximal tubule GLUT 2- Expressed in hepatocytes, intestines, pancreatic -cells

GLUT 3 - Expressed in neurons, testis and spermatozoa - Insulin responsive transporter, found in GLUT 4 myocytes and adipocytes GLUT 5 - found on the apical surface in the intestine and kidney for the transport of fructose. Also found on muscle, adipose tissue, spermatozoa GLUT 7 - It is present in the endoplasmic reticulum membrane and allows efflux of glucose out of the lumen of this organelle after the action of glucose-6-phosphatase on glucose-6-phosphate GLUT1, GLUT3 and GLUT4 also transport dehydroascorbic acid, the oxidized form of vitamin C

Two SGLT isoforms have been identified. SGLT1 Expressed in the late proximal tubule and the intestine This has high affinity for glucose but relative low transport capacity. It transports 2 Na+ and 1 glucose (2:1) SGLT2 Expressed in the early proximal tubule This has a lower affinity but higher transport Capacity.

Facilitated or mediated diffusion differs from simple diffusion in that a) It involves a carrier protein or channel b) It is saturable whereas simple diffusion is not c) It occurs at a faster rate than simple diffusion d) It is a highly specific process e) It exhibits competitive inhibition

The transmembrane channels that permit facilitated diffusion can be opened or closed. They are said to be "gated". Some types of gated ion channels: 1) ligand-gated 2) mechanically-gated 3) voltage-gated

1) Ligand-gated ion channels. Many ion channels open or close in response to binding a small signaling molecule or "ligand". the ligand is not the substance that is transported when the channel opens.

Examples: 1) Acetylcholine (Ach). The binding of the acetylcholine at certain synapse opens channels that admit Na+ and initiate a nerve impulse or muscle contraction. 2) Gamma amino butyric acid (GABA). Binding of GABA at certain synapses designated GABAA in the central nervous system admits Cl- ions into the cell and inhibits the creation of a nerve impulse 3) Second messengers", like cyclic AMP (cAMP) and cyclic GMP (cGMP), regulate channels involved in the initiation of impulses in neurons responding to odors and light respectively.

2) Mechanically-gated ion channels Examples: 1) Sound waves bending the cilia-like projections on the hair cells of the inner ear open up ion channels leading to the creation of nerve impulses that the brain interprets as sound.

2) Mechanical deformation of the cells of stretch receptors opens ion channels leading to the creation of nerve impulses

Active transport
3) Voltage-gated ion channels In excitable cells (neurons and muscle cells), some channels open or close in response to changes in the voltage across the plasma membrane. Example: As an impulse passes down a neuron, the reduction in the voltage opens sodium channels in the adjacent portion of the membrane. This allows the influx of Na+ into the neuron and thus the continuation of the nerve impulse.
Active process describes any process that uses energy to move substances across cellular membranes. The cell uses active transport in three situations: 1) when a particle is going from low to high concentration, 2) when particles need help entering the membrane because they are selectively impermeable, and 3) when very large particles enter and exit the cell. The mechanisms of active transport are of two types depending on whether the movement of solute is linked directly or indirectly to energy-yielding reactions : 1) When the movement is linked directly to energy-yielding reaction, the transport is termed primary active transport (or direct active transport) 2) When the active transport is not directly linked to energy-yielding reactions, the transport mechanism is described as secondary active transport (indirect active transport)

Active processes - require the expenditure of energy by cells:

Active transport = movement of a substance across a cell membrane from an area of low concentration to an area of high concentration using a carrier molecule

1) When the movement is linked directly to energy-yielding reaction, the transport is termed primary active transport (or direct active transport)

2) When the active transport is not directly linked to energy-yielding reactions, the transport mechanism is described as secondary active transport (indirect active transport).

(1) The Na+/K+ ATPase Primary / Direct active transport Ion pumps are the only molecules capable of performing primary active transport Most ion pumps are ATPases, which utilise the energy produced by ATP hydrolysis Example:1. The Na+/K+ ATPase
2. The H+/K+ ATPase 3. The Ca2+ ATPases 4. ABC Transporters { ABC ("ATP-Binding Cassette") }
This is also an example of antiport

It is present in the plasma membranes of most cells and is also known as Na+,K+-ATPase It pumps three Na+ out of the cell and two K+ into the cell for each ATP consumed. Both ions are pumped against their concentration gradient

Secondary active transport

(2) The H+/K+ ATPase
The parietal cells (stomach) use this pump to secrete gastric juice. These cells transport protons (H+) from a concentration of about 4 x 10-8 M within the cell to a concentration of about 0.15 M in the gastric juice (giving it a pH close to 1).
Q : Where else can we find the H+/K+ ATPase pump? A : In the colon and collecting ducts of the kidney.

When transport of a solute is not coupled directly to energy yielding reactions, it is described as secondary active transport It is also called cotransport and uses the energy of the Na+ concentration gradients The sodium moves into the cell and provides the energy to move another substance against its concentration gradient. An example of this is the transport of glucose in the gastro-intestinal tract, and the kidney.

(3) The Ca2+ ATPases

It uses the energy provided by one molecule of ATP to pump one Ca2+ ion out of the cell. In resting skeletal muscle, there is a much higher concentration of calcium ions (Ca2+) in the sarcoplasmic reticulum than in the cytosol. Activation of the muscle fiber allows some of this Ca2+ to pass by facilitated diffusion into the cytosol where it triggers contraction. After contraction, this Ca2+ is pumped back into the sarcoplasmic reticulum. This is done by another Ca2+ ATPase that uses the energy from each molecule of ATP to pump 2 Ca2+ ions.

Secondary active transport

Symport Pumps In this type of indirect active transport, the driving ion (Na+) and the pumped molecule pass through the membrane pump in the same direction. Examples: 1) The Na+/glucose transporter. This transmembrane protein allows sodium ions and glucose to enter the cell together. The sodium ions flow down their concentration gradient while the glucose molecules are pumped up theirs. Later the sodium is pumped back out of the cell by the Na+/K+ ATPase. The Na+/glucose transporter is used to actively transport glucose out of the intestine and also out of the kidney tubules and back into the blood. 2) All the amino acids can be actively transported, for example a) out of the kidney tubules and into the blood b) the reuptake of glutamic acid from the synapse back into the presynaptic neuron by sodium-driven symport pumps.

Characteristics of Facilitated Diffusion & Active Transport - both require the use of carriers that are specific to particular substances (that is, each type of carrier can 'carry' one type of substance) and both can exhibit saturation (movement across a membrane is limited by number of carriers & the speed with which they move materials; see graph below).

3) The Na+/iodide transporter. This symporter pumps iodide ions into the cells of the thyroid gland (for the manufacture of thyroxine) and also into the cells of the mammary gland (to supply the baby's need for iodide).

The transporter categories and arrangement with channels with pumps

Endocytosis and exocytosis These are transport processes that also require energy Endocytosis refers to the process of engulfing material by invaginating the outer membrane of a cell until it buds off within the cytoplasm of the cell as a vesicle containing external material

GLUT transporter

SGLT

Na+-Ca 2+

Think
What is PINOCYTOSIS and PHAGOCYTOSIS?

Answer :
phagocytosis----->intake of solid particles by a cell (phago = cell eat) pinocytosis------>intake of liquid particles by a cell (pino = cell drink) pinocytosis and phagocytosis are 2 types of endocytosis (movement of particles into the cell)

Exocytosis is the reverse of endocytosis (exocytosis-> movement of large amount of material out of cells)

Ionophores
An ionophore is a lipidsoluble molecule usually synthesized by microorganisms to transport ions across the lipid bilayer of the cell membrane.

SUMMARY (1)

SUMMARY (2) :
Cell physiology is the basis of understanding of other major topics including :
Resting membrane potential Action potential in excitable tissues (muscles and neurons) Oral rehydration therapy

Thank you