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Questions and Answers
What is the primary mechanism of solute transport in passive diffusion?
What is the primary mechanism of solute transport in passive diffusion?
Solutes move directly through the lipid bilayer or via aqueous pores without the need for energy.
Explain the difference between primary active transport and secondary active transport.
Explain the difference between primary active transport and secondary active transport.
Primary active transport directly uses ATP to move solutes, while secondary active transport relies on the gradients created by primary active transport.
Describe the role of aquaporins in cell membrane transport.
Describe the role of aquaporins in cell membrane transport.
Aquaporins are water channels that facilitate rapid water transport across the cell membrane.
What physiological example illustrates primary active transport?
What physiological example illustrates primary active transport?
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How do electrical gradients influence membrane transport?
How do electrical gradients influence membrane transport?
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Define osmolality and its importance in physiology.
Define osmolality and its importance in physiology.
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What is the significance of concentration gradients in solute transport?
What is the significance of concentration gradients in solute transport?
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How is water transport across cell membranes regulated?
How is water transport across cell membranes regulated?
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Which ions have significant concentration differences between intracellular and extracellular fluids?
Which ions have significant concentration differences between intracellular and extracellular fluids?
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What transport mechanism would glucose use in facilitated diffusion?
What transport mechanism would glucose use in facilitated diffusion?
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Explain the significance of the Nernst equation in determining the driving force for K+ during an action potential.
Explain the significance of the Nernst equation in determining the driving force for K+ during an action potential.
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How does Fick's Law describe the influence of membrane thickness on the movement of solutes across the membrane?
How does Fick's Law describe the influence of membrane thickness on the movement of solutes across the membrane?
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What role does the lipid-water partition coefficient (Kp) play in the permeability of a solute through the cell membrane?
What role does the lipid-water partition coefficient (Kp) play in the permeability of a solute through the cell membrane?
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Differentiate between active and passive transport mechanisms in terms of energy requirements.
Differentiate between active and passive transport mechanisms in terms of energy requirements.
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What is the role of aquaporins in water transport across the cell membrane?
What is the role of aquaporins in water transport across the cell membrane?
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What are the two forms in which weak acids and bases exist, and how does the pKa relate to these forms?
What are the two forms in which weak acids and bases exist, and how does the pKa relate to these forms?
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Explain how pH trapping affects the diffusion of aspirin across the gastric mucosal barrier.
Explain how pH trapping affects the diffusion of aspirin across the gastric mucosal barrier.
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Describe the role of transmembrane concentration gradients in simple passive diffusion.
Describe the role of transmembrane concentration gradients in simple passive diffusion.
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How does lipid solubility influence drug diffusion across the cell membrane?
How does lipid solubility influence drug diffusion across the cell membrane?
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What are the principal sites of carrier-mediated transport in the body?
What are the principal sites of carrier-mediated transport in the body?
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Differentiate between active and passive transport mechanisms.
Differentiate between active and passive transport mechanisms.
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What role do aquaporins play in water transport across cell membranes?
What role do aquaporins play in water transport across cell membranes?
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How does ionisation impact drug absorption and distribution in the body?
How does ionisation impact drug absorption and distribution in the body?
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What is the main characteristic of molecules that undergo passive diffusion across the cell membrane?
What is the main characteristic of molecules that undergo passive diffusion across the cell membrane?
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Describe how aqueous diffusion differs from facilitated diffusion.
Describe how aqueous diffusion differs from facilitated diffusion.
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What type of transport mechanism requires energy and moves solutes against their concentration gradient?
What type of transport mechanism requires energy and moves solutes against their concentration gradient?
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Explain the process of endocytosis and its significance in cellular transport.
Explain the process of endocytosis and its significance in cellular transport.
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Contrast passive diffusion with active transport regarding energy requirements.
Contrast passive diffusion with active transport regarding energy requirements.
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How does osmolarity affect the movement of water across a cell membrane?
How does osmolarity affect the movement of water across a cell membrane?
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What are the key factors that enable a solute to passively diffuse through a membrane?
What are the key factors that enable a solute to passively diffuse through a membrane?
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What happens to the transport of molecules when there is no concentration gradient?
What happens to the transport of molecules when there is no concentration gradient?
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Define facilitated diffusion and give an example of a molecule that utilizes this mechanism.
Define facilitated diffusion and give an example of a molecule that utilizes this mechanism.
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Study Notes
Membrane Transport
- Key learning outcomes include understanding the regulation of normal membrane function and physiological principles.
- Learning outcomes also include understanding the role of electrical and concentration gradients in solute transport across the membrane.
- Mechanisms of passive diffusion, aqueous diffusion, facilitated diffusion, and active transport should be understood, including primary and secondary active transport.
- Students should be able to provide physiological examples of primary and secondary active transport.
- Recommended reading: Boron, Boulpaep (2017), 'Medical Physiology', 3rd ed.
Total Body Water
- Body fluid compartments for a 70kg adult human are approximately 42 liters.
- Plasma water (3 L):
- [Na+] = 142 mM
- [K+] = 4.4 mM
- [Cl-] = 102 mM
- [protein] = 1 mM
- Osmolality = 290 mOsm
- Interstitial water (13 L):
- [Na+] = 145 mM
- [K+] = 4.5 mM
- [Cl-] = 116 mM
- [protein] = 0 mM
- Osmolality = 290 mOsm
- Intracellular water (25 L):
- [Na+] = 15 mM
- [K+] = 120 mM
- [Cl-] = 20 mM
- [protein] = 4 mM
- Osmolality = 290 mOsm
Ways Small Molecules Cross Cell Membranes
- Passive Diffusion: Directly through the lipid or via aqueous pores (e.g., aquaporins). This method is used by many lipid-soluble molecules.
- Aqueous Diffusion: Through channels in the plasma membrane. This process does not require energy but depends on a concentration gradient.
- Facilitated Diffusion: Uses specialized carrier proteins. Binding and conformational changes facilitate movement across the membrane, but no energy is required.
- Active Transport: Uses specialized carrier proteins to move molecules against their concentration gradient. This process requires energy.
- Endocytosis (pinocytosis): Invagination of the membrane to form a vesicle, encompassing the molecule and transporting it into the cell.
Graphic Summary of Transport Mechanisms
- Passive diffusion occurs down a concentration gradient and is driven by lipophilic molecules.
- Aqueous diffusion occurs via a channel down a concentration gradient.
- Facilitated diffusion involves specialized carrier proteins down a concentration gradient.
- Active transport works against a concentration gradient, requires energy.
Solute Transport Across Cell Membranes
- For passive and aqueous diffusion, the solute moves down its electrical and/or chemical gradient.
- Cell membranes must be permeable to the solute, either through lipophilicity or channels.
Modeling Equations and Concepts
- Electrochemical potential energy difference = Chemical potential energy difference + electrical potential energy difference
- This equation models differences in solute concentration and charged particle concentration across a membrane.
The Nernst Equation
- Describes the equilibrium potential of an ion across a membrane.
- Relevant variables include: membrane potential (Vm), gas constant (R), absolute temperature (T), ionic valence (z), and Faraday constant (F).
- Used to understand the electrochemical gradient of ions.
Calculation of Ek
- Net driving force in volts is Vm - Ex.
- At resting potential, K+ driving force is approximately 17.7 mV (outward).
- At the peak of action potential, K+ driving force is approximately 117.7 mV (outward).
- This correlates with the fluxes of K+ during repolarization.
Diffusion of Electrically Neutral Solutes
- Fick's Law describes the flux (movement rate) of a solute across a membrane.
- Factors determining flux include the solute's lipid-water partition coefficient, diffusion coefficient, and membrane thickness.
- Permeability coefficient integrates these factors.
The Lipid-Water Partition Coefficient
- The lipid-water partition coefficient (Kp or logP) indicates how readily a drug dissolves in a lipid or water environment.
- Hydrophilic molecules have Kp Values less than 1; hydrophobic molecules exceed 1.
Simple Passive Diffusion Example
- Membrane transport of a drug depends on the lipid solubility of the drug (e.g. Ganaxolone, logP = 5.0).
- Lipid insoluble drugs (e.g. Mannitol, logP = 3.1) show different transport patterns across the membrane.
- Transport across the membrane is determined by a concentration gradient.
Application to Pharmacology - pH, Ionisation and Diffusion
- Many drugs are weak acids or bases, existing in ionised and un-ionised forms.
- Ionisation depends on the drug's pKa and local pH.
- The non-ionised form readily penetrates membranes.
Application to Pharmacology - pH Trapping
- Aspirin (acetylsalicylic acid, pKa=3.5) is largely ionized in the plasma (pH 7.4), trapping it in the tissues.
- Drugs with different pKa values exhibit differential absorption patterns.
Principal Sites of Carrier Mediated Transport
- Key sites for carrier-mediated transport include the Blood-brain barrier, Gastrointestinal Tract, Placenta, Renal Tubule, and Biliary Tract.
The Importance of Transporters
- Intestinal solute carrier proteins are crucial for absorbing electrolytes, macronutrients, and micronutrients.
- Examples of drug substrates include B-lactams, angiotensin-converting enzyme inhibitors, and others.
Carrier-Mediated Transport vs Passive Diffusion
- Carrier-mediated transport follows Michaelis-Menten kinetics due to protein involvement.
- Solute transport rate, V, is dependent on maximal transport rate (Vmax) and Michaelis constant (Km) and solute concentration ([S]).
- Passive diffusion does not display saturation kinetics.
Transporters in Facilitated Diffusion
- Hydrophilic molecules require specialized carrier proteins to cross cell membranes.
- Facilitated diffusion does not require energy.
- This process can exhibit saturation kinetics.
The Glucose Transporters
- Glucose transporters (e.g., GLUT1) are members of the SLC2 solute carrier superfamily.
- GLUT1 facilitated diffusion is critical for insulin secretion.
GLUT2 and GLUT5
- GLUT2 and GLUT5 are responsible for facilitated diffusion of fructose and glucose in the gut.
- Glucose and galactose are absorbed via secondary active transport (SGLT1).
- Monosaccharide exit occurs via facilitated diffusion (GLUT2).
Transporters in Active Transport
- Specialized carrier proteins are required for hydrophilic molecule transport against a concentration gradient.
- This process requires energy.
- Carrier-mediated transport shows saturation kinetics.
Primary Active Transport – The Na+/K+ ATPase Pump
- The Na+/K+ ATPase pump is a critical primary active transporter.
- This pump establishes the concentration gradients for secondary active transport mechanisms.
- It moves Na+ and K+ against their electrochemical gradients.
The Na+/K+ ATPase Pump Mechanism
- The pump exists in E1 and E2 conformational states.
- 3 Na+ ions bind intracellularly followed by ATP hydrolysis.
- 2 K+ ions bind extracellularly.
- Conformational changes occur, releasing Na+ and binding K+, releasing the phosphate and returning to E1.
Transport Modes for Na+/K+ ATPase Pump
- The Na+/K+ ATPase operates in normal and reverse modes.
- The pump can facilitate Na+ and K+ exchange, as well as Na+ exchange, and K+ exchange, as separate modes.
Na,K-ATPase Structure
- The Na,K-ATPase is a P-type ATPase with a composition of α and β subunits.
- The α subunit contains ATP-binding sites, while the β subunit is glycosylated.
- Tissue-specific expressions of different alpha subunit isoforms exist.
Evidence for Multiple isoforms of Na/K ATPase
- Different tissues show varying sensitivity to cardiac glycosides, indicating the presence of different isoforms.
- Tissue-specific antibodies against the purified enzyme partially inhibit the Na/K-ATPase activity in other tissues, demonstrating a difference in the isoforms.
Tissue-Specific Expression of Na/K ATPase Isoforms
- The α1, α2, α3, and α4 subunits display unique tissue distributions.
- The α1 subunit is ubiquitously expressed and contains the binding sites for digoxin-like drugs.
Application to Pharmacology – Digoxin
- Digoxin, obtained from foxglove plants, inhibits the Na+/K+ ATPase pump.
- Digoxin binding to the alpha subunit alters intracellular Na+ and Ca2+ levels, influencing cardiac function.
Application to Pharmacology – P-glycoprotein
- P-glycoprotein (P-gp) is a multidrug transmembrane transporter, responsible for drug resistance.
- Key functions include drug elimination via bile transport, kidney excretion, placenta transport, and limiting distribution in the brain.
Primary and Secondary Active Transport Summary
- Primary active transport, such as the Na+/K+ ATPase pump, establishes electrochemical gradients for secondary active transport mechanisms.
- Secondary active transport utilizes electrochemical gradients (e.g., Na+ gradient) to move other solutes.
Na+/glucose and Na+/amino acid Co-transport
- Na+/glucose and Na+/amino acid cotransport mechanisms are critical in postprandial Na+ absorption in the small intestine.
- These co-transport processes are electrogenic via the Na+/K+ ATPase pump.
Na+/H+ Exchange and Absorption
- Na+/H+ exchange (NHE) is essential for Na+ absorption.
- This process occurs at both apical and basolateral membranes with bicarbonate generated by the pancreas driving Na+/H+ exchange in the alkaline apical environment of the intestine.
Na+/H+ and CI-/HCO3 Exchange Summary
- Na+/H+ and Cl−/HCO3− exchange mechanisms occur in parallel in the ileum and proximal colon, contributing to Na+ absorption during the interdigestive period and not postprandially.
- cAMP, cGMP, and Ca2+ regulate NaCl absorption, and reduced absorption can be a cause of secretory diarrhoea.
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