Physiology I 09/13/10 10/08/10 Margaret Anderson

For class PowerPoints, go to www.trinityphysiology.org Tentative schedule F, 09/24 20m QUIZ /discussion EXAM, F 10/08

Physiology I

Physiology is the study of the normal functions of a living organism and its various components. To achieve optimal health, the components must function together.

Cell 1: Homeostasis and membrane function Learning objectives:

Define/describe, explain physiological significance, and give examples: Homeostasis and roles of feedback mechs Distribution of solutes and water in ECF & ICF Cell (plasma) membrane & capillary endothelium Diffusion coefficient (Stokes-Einstein eqn) Permeability, partition coefficient Diffusion (Ficks Law) Carrier-mediated transport Osmosis, osmolarity Osmotic pressure (vant Hoffs Law) Tonicity and effective osmotic pressure Oncotic pressure Selected questions in Cases and Problems 1 & 2, Costanzo 3e.

Claude Bernard (1865) If we break up a living organism by isolating its different parts, it is only for the sake of ease in analysis and by no means to conceive them separately. Indeed, when we wish to ascribe to a physiological quality its value and true significance, we must always refer it to the whole and draw our final conclusions only in relation to its effects on the whole.

Organ systems < organs < tissues < cells

Multicellular organisms require an infrastructure of tissues, organs, and organ systems to ensure survival and functions of individual cells.

DIGESTIVE SYSTEM URINARY SYSTEM CIRC SYSTEM RESPIRATORY SYSTEM

CELLS

Claude Bernard: Constancy of the internal environment is the condition for free life.

HOMEOSTASIS

condition (not static) Adaptive mechanisms respond to conditions or stimuli to produce a relatively constant internal environment Conditions are sensed and then controlled Each system works in concert with others. Homeostasis involves feedback / Walter B. Cannon, the Father of feedforward mechanisms American Physiology, coined the Negative feedback Positive feedback Feedforward mechanisms
word homeostasis in 1929.
Photo: Cannon, W.B. The Way of an Investigator. 1968.

Homeo similar (not same) stasis

Some physiologists argue for using the term homeodynamics instead of homeostasis.

Opposes change: maintains relative status quo

Negative feedback control system

3. Controller compares sensed level with desired level (set point). 4. If a difference, controller sends an error signal to the effector to tell it to bring variable closer to set point: to oppose the change

Desired level of something

Error signal

1. Regulated variable is sensed 2. Sensor feeds back info about its level to the controller

Adapted from Rhoades and Bell, 3rd ed., Fig 1.2

Examples of negative feedback:

Heat/cool a room You eat a candy bar

Negative feedback loops stabilize conditions around a constant value.

Silbernagl & Despopoulos. 2009. Fig D 1, p. 7.

Positive feedback:
Reinforces change escalates a response snowball effect.

Silverthorn 5e, Fig. 6-27b

Positive feedback loops are relatively rare in physiology. : Moves condition away from its initial value

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Positive feedback:
Example: parturition
Reinforces change

Silverthorn 5e, Fig. 6-28

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Anticipates change

Feedforward control

Feedforward controller generates commands without directly sensing regulated variable, although it may sense a disturbance.

Example: smelling or seeing food can stimulate salivation and gastric secretion of HCl.

Feedforward systems often act in concert with feedback systems.

Adapted from Rhoades and Bell, 3rd ed., Fig 1.2

Homeostasis of the internal environment: Intracellular fluid (ICF) Extracellular fluid (ECF)
DIGESTIVE SYSTEM URINARY SYSTEM CIRC SYSTEM RESPIRATORY SYSTEM
CELLS

ICF and ECF reach a state of [dynamic] osmotic equilibrium, but they are in chemical and electrical disequilibrium

Healthy humans maintain remarkably constant conditions in their blood and tissue fluids

What is the weight of interstitial fluid in this person? What is the volume of interstitial fluid in this person?
(2 subcompartments)

Body fluid compartments (70 kg person)

TOTAL BODY WATER (~45 L) ECF (15 L) cell membrane capillaries

INTRACELLULAR FLUID (~30 L) Contained within cells

INTERSTITIAL FLUID (~12 L) Bathes cells

no protein: ultrafiltrate of plasma

PLASMA (~3 L) Liquid part of blood inside vessels

Contains proteins e.g. albumin, clotting proteins

Explain: Total blood volume is ~ 5 L. Plasma is ~ 3 L.

See Costanzo 3e and 4e Figure 1-1.

We need to consider the mechanisms by which substances move (or not) across the cell membrane and the endothelium .
Screen 51 shows c.s. of capillary

Cell membrane: simple diffusion and carrier-mediated transport. Endothelium: bulk flow dependent on oncotic pressure and blood pressure.
Endothelial cells make up the capillary wall (endothelium) & separate blood plasma from interstitial fluid.

interstitial fluid

Silverthorn 5E, fig. 3-25a

The intracellular environment is in chemical and electrical disequilibrium with the interstitial fluid.
FACTS : 1. The major cation in the ICF is K 2. The major cation in the ECF is Na 3. The major anions in the ICF are large proteins that carry a net negative charge 4. The major anion in the ECF is Cl

5. 6.

Cells have a shell of negative charges inside and positive charges outside Except for the shell, the ICF and ECF are electroneutral

+ + ++ + + + + + + + + + + + +

Intracellular (C), interstitial (I) and plasma (P) compartments are in chemical disequilibrium

ECF ICF
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FYI: The most abundant nonionic small molecule solutes in the ECF are glucose and urea.

Silverthorn, 5E, Fig 5-3b.

The cell membrane consists of a phospholipid bilayer with associated proteins and carbohydrates
Structural components of the cell membrane determine the movement of materials into and out of the cell.

5-10 nm

cholesterol

Phospholipid molecules are amphipathic: polar (hydrophilic) heads and nonpolar (hydrophobic) fatty acid tails.

Silverthorn 5e Fig. 3-6

Silverthorn 5e Fig. 2-8

Materials pass through the membrane by simple diffusion or by carrier-mediated transport

The cell membrane is selectively permeable, which means that some substances can pass through it and others not.
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Animal Physiology 2e, Fig 2.1

Examples: Diffusion and carrier-mediated transport across membranes

Diffusion downhill along conc gradient through lipid bilayer or through AQP channel

Leucine transporter (LeuT) depends on Na gradient to move leucine uphill, from low concentration to high concentration. The Na gradient is set up initially by expenditure of ATP.

Maltose ABC transporter depends on direct use of ATP as source of cellular energy

Wang, y., S.A. Shaikh, and E. Tajkhorshid. 2010. Exploring transmembrane diffusion pathways with molecular dynamics. Physiology 25: 142-154.

Membrane transport molecules

Channels (simple diffusion) Movement along concentration gradient (downhill)

Carriers

Passive transporters (facilitated diffusion no ATP used)

Primary active transporters (use ATP directly)

Secondary active transporters (couple to ion gradients set up by primary active transport)

Movement against concentration gradient (uphill)

Simple diffusion results from kinetic energy of molecular motion. Rules: 1. Diffusion does not use energy from an outside source. It is referred to as passive transport. 2. Molecules move from [high] to [low], downhill, along a concentration gradient 3. Net movement occurs until the concentrations come to equilibrium 4. Diffusion can take place in an open system or across a partition 5. Diffusion is rapid over short distances but slow over long distances 6. Diffusion rate increases with increased temperature 7. Diffusion rate increases with a greater concentration gradient 8. Diffusion rate is inversely proportional to molecular size

Time required for diffusion increases exponentially with the distance traveled (t ~ x2). E.g., a molecule that travels one m in 0.5 ms will travel 100 m in 5 s: 1m ~ (12) x (0.5 x 10-3 s) = 0.5 x 10-3 s 100 m ~ (1002) x (0.5 x 10-3 s) = (10 x 103) x (0.5 x 10-3 s) = 10 x 0.5 s = 5 s

Molecular agitation

Some materials pass through the bilayer by simple diffusion

partition coefficient (K) diffusion coefficient (D)

(CA CB)

Silverthorn 5e, Fig. 5-6.

(CA CB) x

Costanzo, p. 7

J = PA (CA CB)

P = KD/x

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partition coefficient: K = conc in oil/conc in water

diffusion coefficient (D): Stokes-Einstein eqn

D=

KT 6r

(CA CB)

Silverthorn 5e, Fig. 5-6.

(CA CB) x

Costanzo, p. 7

J = PA (CA CB)

P = KD/x

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Of these four solutes, which has the highest permeability in a lipid bilayer? Which has the lowest permeability?
Table 1-1 (Costanzo Cases, 3e) Solute A B C D Molecular radii and oil-water partition coefficients of four solutes Molecular radius, 20 20 40 40 Oil-water partition coefficient, K 1.0 2.0 1.0 0.5

Lipid bilayer, surface area = 1 cm2 , permeability = 5 x 10-5 cm/sec

Solute A: 20 mM/ml

Solute A: 10 mM/ml

Calculate the net rate of diffusion of Solute A across the lipid bilayer. Which equation will you use? In which direction will net diffusion occur?

See Case 1, question 6, Costanzo Cases and Problems, 3e.

Materials pass through the membrane by simple diffusion or by carrier-mediated transport

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1. Simple diffusion through phospholipid bilayer or channel (passive transport) 2. Carrier-mediated transport a. facilitated diffusion (passive transport) b. primary active transport c. secondary active transport

Facilitated diffusion is an example of carrier-mediated transport Like simple diffusion: No outside source of energy is used Direction of transport is from [high] to [low] Net transport stops when concentrations of the molecule are equal on both sides of the membrane
Facilitated diffusion carrier proteins are often called passive transporters.

The carrier protein does not form an open passage between the ICF and ECF
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Silverthorn 5e, Fig 5-11

Like other carrier-mediated transport systems, facilitated diffusion exhibits: Stereospecificity Saturation Competition

All transport molecules* exhibit stereospecificity

The binding site recognizes, binds and transports only a specific molecule (or subset of molecules) For example, the carrier for D-glucose (GLUT) will bind and transport D-glucose but not the nonphysiological stereoisomer L glucose.

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Silverthorn 5e, Fig 5-11

*Transport molecules involved in facilitated diffusion, primary active transport and secondary active transport all exhibit stereospecificity, saturation, and competition.

All transport molecules exhibit competition

The GLUT transporter binds and transports both glucose and galactose, which compete for the glucose binding site. In the presence of galactose, the transporter moves fewer glucose molecules per unit time across the membrane because it carries galactose some of the time.
Glucose only

Glucose in presence of 1 mM galactose

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Silverthorn 5e, Figs 5-11 and 5-17

All transport molecules exhibit saturation

When all carrier molecules of a given type are bound with substrate molecules, the population is saturated.
The rate of transport stays the same (at its maximum) once all carrier molecules are occupied.

The rate of transport is proportional to the [substrate]until all carrier molecules are transporting substrate.

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Silverthorn 5e, Figs 5-11 and 5-19

The Na-K ATPase pump : an example of primary active transport.

This carrier-mediated transport requires the direct input of energy from ATP. The carrier molecule moves Na and K ions uphill, against their concentration gradients.

E1

E2

Na-K ATPase pump

Costanzo, 3e and 4e, Figure 1-6

Examples of other primary active transport systems:

Cell (plasma) membrane Ca ATPase (PMCA) pumps Ca ions out of cell most cells Sarcoplasmic and endoplasmic reticulum Ca ATPase (SERCA) pumps Ca out of cytoplasm into the sarcoplasmic reticulum (or endoplasmic reticulum) muscle and some other cells. H-K ATPase pumps H from the ICF to the lumen of stomach (parietal cells in the gastric mucosa)

Secondary active transport couples the movement of solutes across the

membrane. This carrier-mediated transport depends on the indirect utilization of ATP for energy. Co-transport (symport) Counter-transport (antiport)

Facilitated diffusion

Na and glucose move in the same direction (into the cell): symport. This SGLT transporter moves glucose from [low] outside to [high] inside against its concentration gradient.
Constanzo 3e and 4e, Figs 1-7 and 1-8.

Na and Ca move in opposite directions (Na into the cell and Ca out): antiport. Ca is moved from [low] inside to [high] outside against its concentration gradient. In both examples the potential energy stored in the Na concentration gradient is used to drive the carrier. ATP was used indirectly to maintain the Na concentration gradient.

Review and anticipation

1

Question:
Which of these transport mechanisms [1], [2], [3] would be inhibited by a cardiac glycoside such as ouabain?
2

5
Fig 1-8

Osmosis and tonicity

In osmosis, water flows across a semipermeable membrane from a solution with low [solute] to a solution with high [solute]. We will now address solute concentrations and the movement of water.
About 2/3 of the bodys water is contained in cells. The rest is distributed between the interstitial fluid and blood plasma.

Animal Physiology 2e Fig 26-1

Clinicians estimate a persons fluid loss in dehydration, for example, by equating weight loss to water loss. Water loss/gain will affect solute concentrations.

Osmolarity
Osmolarity is expressed as the concentration of osmotically active particles (ions or intact molecules) in a liter of solution (Osm/L) Osmolarity = g x C Osmolarity (Osm/L) = g (# particles/mol in Osm/mol) x C (concentration in mol/L) 1 Osmole = 6x1023 osmotically effective entities e.g. Glucose (does not dissociate in soln): (6x1023 particles of gluc / 6x1023 molecules of gluc) = 1 OsM/mol gluc for 1 mol/L glucose: Osmolarity = 1 OsM/mol x 1 mol/L = 1 OsM/L e.g. NaCl (assume complete dissociation into Na and Cl) [(6x1023 Na part) + (6x1023 Cl part)] / 6x 1023 NaCl molec) = 2 OsM/mol NaCl for 1 mol/L NaCl: Osmolarity = 2 OsM/mol x 1 mol/L = 2 OsM/L

Osmolarity also expressed as Osmolarity = n x C where n is the number of dissociable particles per molecule

Comparing osmotic concentrations

SOLUTION A = 1 OsM/L Glucose SOLUTION B = 2 OsM/L Glucose SOLUTION C = 1 OsM/L NaCl

Isosmotic: two solutions have the same osmotic concentrations (e.g. A and C) Hyperosmotic: a solution with a higher osmotic concentration than the one to which it is compared (e.g. B is hyperosm to A & C) Hyposmotic: a solution with a lower osmotic concentration than the one to which it is compared (e.g. A & C are hyposm to B) What is the molar concentration of Soln A? Soln B? What is the approximate molar concentration of Soln C?

Osmolarity and osmolality

Osmolarity Generated by Temperature Units Number of molecules dissolved in 1 L of solvent Affects volume of solvent Osm/L or mOsm/L Osmolality Number of molecules dissolved in 1 kg of solvent Does not affect mass of solvent Osm/kg or mOsm/kg

Osmolality is the preferred term for physiological systems. Physiological solutions are dilute (usually expressed in mOsm/L or mOsm/kg), and the solvent is water.

PROBLEM: Osmotic concentration (osmolar, Osm/L; milliosmolar, mOsm/L) is the sum of the molar concentrations of all undissociated molecules, anions, and cations. Give the osmolarity of the following:

100 mM/L NaCl = ________ mOsm/L 100 mM/L K2SO4 = ________ mOsm/L 100 mM/L CaCl2 = ________ mOsm/L 100 mM/L glucose = ________ mOsm/L 100 mM/L glucose + 100 mM/L NaCl = _______ mOsm/L

Substance Na+ K+ ClHCO3Ca++ Phosphate (Pi) NH4+ NaCl CaCl2 Glucose Urea

Atomic/Molecular Weight Equivalents/mol Osmoles/mol 23.0 39.1 35.4 61.0 40.1 95.0 18.0 58.4 111 180 60 1 1 1 1 2 3 1 2* 4 1 1 1 1 1 1 1 2 3 1 1

Concentrations of ions may be expressed in equivalents per liter. An equivalent (eq) is the molarity of an ion times the number of charges it carries.

*One equivalent each from Na+ and Cl-.

does not dissociate completely in solution. The actual osmoles/mol is 1.88. However, for simplicity, a value of 2 is often used. Ca++ contributes two equivalents, as do each of the 2 Cl- ions.

NaCl

Berne & Levy 6e, Table 1-4

Silverthorn 5e, Fig 2-14

Problem: Atomic /molecular mass

Which of these solutes dissociate(s) when dissolved in water, and into what?

Each of these molecules is made into a 100 millimolar soln. Give the mEq/L concentration of each component NaCl: Na+ ___ mEq/L Cl- ___ mEq/L Cl- ___ mEq/L CaCl2: Ca2+ ___ mEq/L K2SO4 K+ ___ mEq/L SO42- ___ mEq/L

NaCl CaCl2 K2SO4 Urea, (NH)2CO Glucose, C6H12O6

Osmosis occurs when water moves across a membrane from a dilute soln of solute to a more concentrated soln, until the concs are equal.

The concentration difference produces an Osmotic pressure exerted by a solute osmotic pressure difference, which is the is the driving force for osmosis. driving force for osmosis.

Gauge measures pressure in atm or mm Hg

Costanzo 3e and 4e, Fig 1-9

Osmotic pressure is calculated by vant Hoffs Law

= nCRT where n = number of dissociable particles per molecule C = total solute concentration R = gas constant (0.082 atm L/mol oK) T = temperature in degrees Kelvin

A B

Consider a solution of urea: 1 mmol/L @ 37oC What is its osmotic pressure () expressed in atmospheres? Expressed in mm Hg? Assume semipermeable membrane permeable only to water.

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Silverthorn Fig 5-26 (3)

Osmotic pressure is calculated by vant Hoffs Law

= nCRT where n = number of dissociable particles per molecule C = total solute concentration R = gas constant (0.082 atm L/mol oK) T = temperature in degrees Kelvin

A B

Consider a solution of urea: 1 mmol/L @ 37oC What is its osmotic pressure () expressed in atmospheres? Expressed in mm Hg? Assume semipermeable membrane permeable only to water.
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n=1 C = 0.001 M/L = 1 mM/L 37oC = 310o K R = 0.082 L-atm * mol-1 * K-1 RT = 25.45 L-atm/mol = 2.54 x 10-2 atm = 19.3 mm Hg

Tonicity of the solution is described relative to the cells response

Lang, F. and S. Waldegger. Amer Sci 85: 456 463. 1997.

Soln:

hypertonic

isotonic

hypotonic

very hypotonic

TONICITY is defined biologically in terms of the response of a living cell immersed in a solution

_____tonic _____tonic _____tonic

Tonicity and osmolarity (osmolality) are both taken into account to determine the appropriate intravenous solution to administer to a patient. Copyright 2010 Pearson Education, Inc.

Cell membranes are variably permeable to substances The reflection coefficient (, sigma) is a measure of the ability of a molecule to pass through the membrane
=1 = 0 to 1 =0

Impermeable e.g. serum albumin

partially permeable

completely permeable e.g. urea

Vant Hoffs eqn modified by Staverman: = (nCRT)

Constanzo 3e & 4e, Fig 1-10

Consider two solutions: 300 mmol/L sucrose and 300 mmol/L urea O What is the osmotic concentration (osmolality) of each? ll Are they isosmotic? Are they isotonic? H2N C NH2 The cytoplasm of red blood cells is ~ 300 mOsm/kg H2O RBCs in sucrose soln maintain normal volume
=1 =0-1 =0

Explain results
RBCs in urea soln swell and burst = (nCRT)
If a molecule exerts osmotic pressure across a membrane, it must not cross the membrane.

RBC membrane is permeable to urea. Urea has a reflection coefficient (, sigma) of 0. Therefore urea does not exert any effective osmotic pressure. Water follows urea into cell along osmotic gradient. Cell swells and bursts. RBC membrane is impermeable to sucrose ( = 1). Sucrose is an effective osmole because it balances osmotic pressure of the intracellular solutes.

Practice: p. 7, Costanzo Cases 3e #4. Calculate the osmolarity and effective osmotic pressure of each solution at 37oC, RT = 25.45 L-atm/M, or 0.0245 L-atm/mM. Then answer #5-#7.
Table 1-2 (Costanzo Cases 3e) Solution 1 2 3 4 5 6 Comparison of six solutions

Solute Urea NaCl NaCl KCl Sucrose Albumin

Concentration g = n 1 mM/L 1 mM/L 2 mM/L 1 mM/L 1 mM/L 1 mM/L 1.0 1.85 1.85 1.85 1.0 1.0

0 0.5 0.5 0.4 0.8 1.0

g = n, osmotic coefficient; , reflection coefficient

Oncotic pressure is osmotic pressure produced by large molecules (especially proteins)

Oncotic pressure is produced by large proteins in the plasma (=colloid osmotic pressure). Plasma oncotic pressure combines with the hydrostatic effects of blood pressure to influence the movement of fluids across capillary walls. Copyright 2010 Pearson Education, Inc.

contains protein molecules contains no protein molecules

Silverthorn, 5e. Fig. 5-3.

Capillary endothelial cell

lumen

Water and dissolved substances

vesicular transport of some proteins

*** *** *** *** *** ~ -25 *** cap plasma ***
proteins

water-filled pore lipid-soluble substances e.g. CO2, O2

BP>25 net filtration out of cap BP<25 net absorption into cap

BP

cap ~ 25 mm Hg

hydrostatic PIF ~ 0

water-soluble substances e.g. Na+, K+, glucose, aas