Membrane Physiology and Epithelial Self Study Frequently Asked Questions 3. How do you differentiate osmolarity and tonicity?

How can a solution initially be hypo-osmotic but also isotonic? Isotonic means the cell volume doesnt change in steady-state when you expose it to the solution. Iso-osmotic means that the solution has the same water concentration (mOsmoles) as some reference solution, usually the intracellular water in our case. Yes, you can have a hypotonic solution that is iso-osmotic. For instance, say X is permeable to the membrane and Y isnt. I have 100 mOsm of X and 100 mOsm of Y inside the cell and I put it in a very large volume of 50 mOsm X and 100 mOsm Y. The new solution is hypo-osmotic relative to the intracellular solution (150 mOsm < 200 mOsm). But X can diffuse across the membrane. So X diffuses out of the cell until the concentration of X both inside and outside is 50 mOsm (remember that the extracellular solution is a very large volume). Now the new steady-state has 150 mOsm inside and 150 mOsm outside. So the solution is isotonic (i.e. it ultimately causes no change in cell volume). As far as transient changes go, consider the same problem. At the instant I add the new solution to the cell, there is a water gradient across the membrane. So initially, only at the very beginning, the cell will swell (water will come in down its gradient). But this is only a transient change because eventually enough of X moves out of the cell to stop the swelling, then, eventually, to bring the volume back to where it started. 4. Im having trouble with the problem below. Why doesnt the cell swell? A cell with no membrane permeability to any solutes, but permeable to water is placed in a solution of 225mM NaCl. The cell initially had 300mM solute concentration and a volume of 1.0. When a new steady state is achieved the cell's volume will be: (The answer is 0.67) The key is realizing that Na and Cl dissociate to form two separate ions and therefore two separate particles. This makes the osmolarity outside the cell 450 mOs not 225. So we have 450 milliosmolar outside and 300 inside. Water flows down its gradient from outside to inside, shrinking the cell until you have 450 milliosmoles/L both inside and outside: 300 milliosmoles/L (1 L) = 300 milliosmoles in the cell 450 milliosmoles/L (x) = 300 milliosmoles X = 300 milliosmoles/(450 milliosmoles/L)= 2/3 L 5. From an old test: Red blood cells (RBC) with an initial volume of 1 liter are placed in a large beaker containing 2 L of 300 mM NaCl solution. Assume that the RBC initially contains 300 mOsm/L of impermeable solutes. Additionally assume that the RBC membrane is impermeable to both Na+ and Cl-. Once placed in the beaker, water will move across the RBC membrane until their total volume becomes: a. 0.5 L b. 0.6 L c. 0.75 L d. 1.0 L (i.e., no volume change) e. 1.67 L

Guess I am still having trouble with osmo problems. Here the NaCl dissociates so there is 600mM of solute, so water will flow out of RBC. Using the concentration equation, I get there is no volume change. What gives? Answer is b. The question is tricky. The final osmolarity inside and outside will be the same but water is moving from one space to another so the volume is changing on both sides of the membrane. Since the final concentration on both sides of the membrane must be the same we calculate the total concentration of solute and figure that this sill be the concentration everywhere at steady- state: 600 mOsmoles/L (2L) + 300 mOsmoles/L (1 L) = 1500 mOsmoles total 1500 mOsmoles/3 L = 500 mOsmoles/L If the concentration inside need be 500 mOsmoles/L then: 300 mOsmoles/L (1 L initial volume) = 300 mOsmoles within the RBC 500 mOsmoles/L (X L new volume) = 300 mOsmoles X = 300/500 = 0.6 L 6. I just completed the first half of an old test and noticed that there were quite a few questions regarding "clinical correlations" from our syllabus. I am concerned about this as I did not focus on those while doing the reading...will there be the same proportion of those on the actual exam? I will not consciously add any questions directly from clinical correlations unless it was covered in a workshop or is a block case (like sickle cell anemia).

If you find any questions from this lecture or the self-study on the exam, you have a legitimate complaint and you may bring it to me to discuss it. Please be warned, however, that if it is something which you should have been able to figure out from the regular material, I probably wont recommend that the points be given. Having said that, it should not be necessary for you to directly study these sections. For instance, I would never just assume that you knew what a disease was from one of these sections and what causes it. But I might introduce the disease as an example of nbormal function gone awry, give you all the information you need, then expect you to figure out the answer to the problem. It wouldnt hurt to look at these sections if you have the time but you should not under any circumstances need to know them going in to answer my questions. You certainly should not concentrate on them.

PRACTICE EXAM QUESTIONS WITH EXPLANAITONS These questions reflect the level of difficulty which you will find in many if not most of the exam questions relevant to this lecture, the computer workshop and the self study. I have provided explanations for the answers. 1. Red blood cells (RBC) with a total intracellular fluid volume of 2 L are place in a bucket containing 4 L of a 100 mM NaCl aqueous solution. Assume for the purpose of this question only that the RBC initially contain 400 mOs/L of impermeant solutes. Also assume that the RC membrane is impermeable to Na+ and Cl-. Water will move across the membrane of the RBC until their intracellular volume becomes: a. 1L b. 1.5L c. 2.5L d. 3L e. 4L

1. Answer = d. A 100 mM NaCl solution has an osmolarity of 200 mOs/L. 4 liters of 100 mM NaCl solution contains a total of 800 mOsmoles of solute. 2 liters of RBC (with 400 mOs/L of impermeant solutes) contain a total of 800 mOsmoles of solute. Thus there is a total of 1600 mOsmoles of solute in a total volume of 6 liters (RBC initially have 2 L plus 4 L of NaCl solution). The final osmolarity of the RBC and the solution will be 1600 mOs/6 L = 266.67 mOs/L. Since the membrane is impermeable to both Na+ and Cl-, only water can cross the RBC membrane. The amount of solute in the RBC will not change, so water must flow until their osmolarity is 266.67 mOs/L. Remembering that concentration = amount/volume (so volume = amount/concentration) it can be calculated that the new volume must be 800 mOsmoles/266.67 mOs/L = 3 L.

Old Exam Questions 5. A hypothetical cell contains 300 mOm/L of X and 300 mOsm/L of Y. The cell membrane is permeable to Y but not permeable to X. You are holding a bag of solution with 600 mOsm/L of Y and no X. Relative to the hypothetical cell, the solution in the bag is: a. hypo-osmotic and isotonic b. hyper-osmotic and hypertonic c. hypo-osmotic and hypotonic d. iso-osmotic and hypertonic e. iso-osmotic and hypotonic 8. The lipid bilayer portion of most membranes usually is: a. highly permeable to ions and polar solutes b. permeable to most dissolved gases c. completely hydrophobic d. completely hydrophilic e. both a and d are correct a. 9. Diffusion: is an active process that can build up unequal transmembrane concentrations of an uncharged solute that is permeable across a membrane b. proceeds at the same rate for all solutes c. moves smaller uncharged solutes more rapidly that larger uncharged solutes d. will move a given solute 10 times further (on average) in 10 times more time e. none of the above are correct 11. The lipid bilayer portion of the membrane is: a. highly permeable to ionic and plar solutes b. completely impermeable to all solutes c. completely impermeable to dissolved O2 and CO2 d. generally permeable to non-polar (hydrophobic) solutes e. highly permeable to water and all solutes 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. Answers: C C B A E B D B C D D

BODY FLUIDS - STUDY QUESTIONS 3. Distinguish between "tonicity" and "osmolality." Illustrate the lack of identity between "isotonic" and "isosmotic" by an example in which the two clearly are different. 5. A 70 kg male has a body fluid osmolality of 290 mOsm/kg water. Assume 290 millimoles of NaCl are added to the ECF. Describe in words what fluid and solute shifts, if any, take place. Calculate the final osmolality of the body fluids, and the final ECF and ICF volumes. 6. A 70 kg male has a body fluid osmolality of 290 mOsm/kg water. Assume 2 liters of isotonic saline are added to the ECF. Describe in words what fluid and solute shifts, if any, take place. Calculate the final osmolality of the body fluids and the final ECF and ICF volumes. 7. A patient on the renal ward weighed 100 kg, had a plasma osmolality of 290 mOsm/kg water, and the following volume distribution: TBW 60% body weight, ICF volume 40% body weight, and ECF volume 20% body weight. After drinking 5 liters of water rapidly, and following complete absorption but prior to any excretion, what would be the new ICF and ECF volumes (in liters)? 8. In the patient in 7, what would be the new plasma osmolality in mOsm/kg water?

1.

In the diagram below are two solutions (A and B) separated by a membrane permeable to water but not solutes. Use this as the starting condition for the questions below.

A.
480 osmoles 10 liters
a.

B.
252 osmoles 6 liters

In this initial condition, what is the osmolarity of solution A? In this initial condition, what is the osmolarity of solution B?

b. c. d. e. 2. a. b. c. d. e. 3. a. b. c. d. e. 4. 5.

If you compare osmolarities of solution A to B, solution A is ___________ to B. In which direction will water move between these compartments? After all osmosis has occurred, what will the osmolarity of compartment A be? How much total water has moved into or out of compartment A? If a 5 liter solution with 20 osmoles of sucrose (A) is separated from a 2 liter solution with 10 osmoles of sucrose (B) by a membrane impermeable to sucrose and permeable to water, in which direction will movement of water occur? from solution B to solution A from solution A to solution B (A=4 OsM, B=5 OsM) to solution A and then back to solution B to solution B and then back to solution A there will be no movement of water in this case A 300 mOsM urea solution is __________ hyposmotic to a 200 mM solution of NaCl. (200mM NaCl = 400 mOsM) hyperosmotic to a 200 mM solution of NaCl. isosmotic to a 200 mM solution of NaCl. isosmotic to a 600 mM solution of NaCl. isosmotic to a 0.18 % solution of NaCl. What are the two extracellular fluid compartments in the body? In the diagram below are ECF and ICF compartments separated by a membrane permeable to water and to K+ Use this as the starting condition for the questions below.

ECF
added solution: 10% KCl
a. b.

ICF
7800 mosmoles 30 liters

3900 mosmoles 15 liters

What is the osmolarity in the ICF before any KCl solution is added? One liter of a 10% potassium chloride solution will be added to the ECF. What is the osmolarity of the KCl solution? (molecular weight of K= 39.1, Cl = 35.5) (KCl has a disassociation constant of 2 meaning each mole becomes 2 osmoles) The KCl solution is _____________________to the body (ECF and ICF). circle your answer: hyposmotic isosmotic hyperosmotic After the KCl solution is added, how does the osmolarity of the body change? circle your answer: osmolarity decreases no change osmolarity increases

c. d.

e.

The K+ ions diffuse through an open ion channel into the ICF but the Cl- ions cannot move into the ICF. Which compartment has a higher concentration of non-penetrating solutes after the addition of the KCl solution (before water or solutes move)? the ECF they are the same the ICF

f.

Circle your answer for which way net water movement will be after the KCl addition: into ECF no net water movement into ICF

g.

Based on your answer directly above, what is the tonicity of the KCl solution?

6.

A wilderness firefighter has just completed 12 hours of fighting a forest fire. She followed procedure carefully by drinking water or gatorade every hour for the 12 hours but towards the end of those 12 hours she overheated and vomited twice, losing both fluid and ions. After wards, she exhibited signs of a possible osmotic imbalance. Answer the following questions based on this situation: After vomiting, she was treated by local paramedics who gave her 500 ml of water to drink. By drinking the water did the firefighter increase or decrease her body osmolarity? At the end of the 12 hours and even after drinking the water, she continued to exhibit symptoms of a possible osmotic imbalance. She was taken to the hospital where the measures shown on the chart below were determined. Fill in the remainder of the chart. (Be sure to include units.)

a.

b.

volume solute amount osmolarity

Total body 30 liters

ECF 10 liters

ICF 20 liters

c.

260 mOsM

Based on the information above, the firefighter is hyposmotic, isosmotic or

hyperosmotic relative to normal? d. The emergency room health care team disagrees about which IV solution to give the firefighter. Her osmolarity is low, and her body weight and blood pressure indicates that she is a little dehydrated (needs some fluid in cells). Refer to the list of IV solutions on page 156, Table 5-9. For a normal person, circle your answers below to show how body osmolarity changes (increase, no change or decrease) and which way water moves when each IV fluid is added to the body. Normal person (OsM=285-295 mOsM, isomotic to normal saline and to 5 % glucose) IV Fluid Change in OsM? Water movement? . 0.9% NaCl (normal saline) N/C into ECF N/C 0.45% NaCl N/C into ECF 5% glucose in 0.45% NaCl N/C into ECF N/C 5% glucose in 0.9% NaCl N/C into ECF N/C Which IV fluid would you choose for the firefighter and why?

into ICF N/C into ICF into ICF into ICF

1.

A.
480 osmoles 10 liters
a. 48 OsM 42 OsM b. c. d. hyperosmotic. From B to A.

B.
252 osmoles 6 liters

480 + 252) / (10+6) = 732/16 = 45.75 OsM

Note that you can solve this knowing that each compartment will be equal in the end. Find the osmolarity of A+B; it will = A or B. e. 480 osmoles/ x liters = 45.75 OsM x = 10.49 L 10.49-10 = 0.49 L

0.49 L added Solve knowing what the final osmolarity will be and knowing the number of osmoles. Subtract final volume from the initial volume to find how much added. This can be checked by solving for compartment B also to make sure the same amount leaves B to have osmolarity = 45.75 2. a. from solution A to solution B (A=4 OsM, B=5 OsM) * If the membrane was permeable to sucrose, the answer would be e. 3. a. hyposmotic to a 200 mM solution of NaCl. (200mM NaCl = 400 mOsM) 4. 5. plasma and interstitial fluid

In the diagram below are ECF and ICF compartments separated by a membrane permeable to water and to K+ Use this as the starting condition for the questions below.

ECF
added solution: 10% KCl
h. i. convert % to g/L: convert g/L to osmol/L: 260 mOsM

ICF
7800 mosmoles 30 liters

3900 mosmoles 15 liters

10% = 10 g/100ml = 100g/L 100g/L (1mole/74.6g) (2osmole/1mole) = 2.68 OsM

j. hyperosmotic k. l. Osmolarity increases The ECF

m. Into ECF n. hypertonic

* You could set up this problem using a penetrating solute to show that you can have a hyperosmotic solution that is hypotonic.

6.

A wilderness firefighter has just completed 12 hours of fighting a forest fire. She followed procedure carefully by drinking water or gatorade every hour for the 12 hours but towards the end of those 12 hours she overheated and vomited twice, losing both fluid and ions. After wards, she exhibited signs of a possible osmotic imbalance. Answer the following questions based on this situation: a. decrease (made more dilute with water, osmolarity = zero) b.

volume solute amount osmolarity c. d. hyposmotic

Total body 30 liters 7800 mosmoles 260 mOsM

ECF 10 liters 2600 260

ICF 20 liters 5200 260

The emergency room health care team disagrees about which IV solution to give the firefighter. Her osmolarity is low, and her body weight and blood pressure indicates that she is a little dehydrated (needs some fluid in cells). Refer to the list of IV solutions on page 156, Table 5-9.

For a normal person, circle your answers below to show how body osmolarity changes (increase, no change or decrease) and which way water moves when each IV fluid is added to the body. Normal person (OsM=285-295 mOsM, isomotic to normal saline and to 5 % glucose) IV Fluid Change in OsM? Water movement? . 0.9% NaCl (normal saline) N/C into ECF N/C 0.45% NaCl N/C into ECF N/C into ECF N/C 5% glucose in 0.45% NaCl 5% glucose in 0.9% NaCl N/C into ECF N/C

into ICF N/C into ICF into ICF into ICF

Which IV fluid would you choose for the firefighter and why? 5% glucose in 0.45% NaCl. It will increase her osmolarity and move water into her cells.

APPENDIX: Some Volume Regulation Study Questions: You are not specifically responsible for the precise details of the discussion in this Appendix. However, you are responsible for the various messages that it conveys and the general problemsolving techniques it presents. So, a few examples may help clarify the points (or rules) a-d listed on pages 20-21.

First consider a cell that is placed in a large volume of solution (sufficiently large that changes in cell volume and concentrations do not significantly affect the concentration or volume of the extracellular solution). Consider that the cell contains two classes of solutes (particles) which will be called X and Y. The membrane is permeable to X, but impermeable to Y. The initial concentration (here concentration and osmolarity are equivalent) of X is 200 mM (or mOsm/L) and [Y]i = 100 mOsm/L.

Question 1:

If the external solution also contains only types X and Y with the following osmolarites [X]e = 100 mOsm/L,

[Y]e = 100 mOsm/L, what will happen to cell volume in steady state and transiently?

Answer: In the steady state there will be no change in cell volume. The final concentration of X inside the cell ([X]i) will eventually become 100 mOsm/L, but this will not occur by diluting of X due to cell swelling; instead it will result from X leaving the cell. Transiently the cell will briefly swell because the solution is hypo-osmotic, but this swelling will disappear as X leaves the cell. Note that in the steady state hypo-osmotic is not necessarily the same as hypotonic. This solution was initially hypoosmotic, but isotonic. In the steady state the solution and the cells intracellular fluid will be isotonic and iso-osmotic

Question 2:

If the solution contains [X]e = 100 mOsm/L and [Y]e = 200 mOsm/L, what will happen to the volume of the

cell in the steady state relative to its initial volume (assume that cell concentrations to begin with are the same as those described originally).

Answer: The final volume will be of the initial volume. This will raise [Y]i to 200 mOsm/L (remember that Y can not cross the membrane, so reducing cell volume by 50% will double the concentration of Y). Of course X can cross the membrane, so eventually (i.e., in steady state) [X]i will be 100 mOsm/L, which is the same as [X]e. Transient changes are harder to predict in this situation (so dont worry about it). Note that in this case the solution was initially iso-osmotic, but hypertonic.

Question 3: What will happen to cell volume if the cell is placed in an iso-osmotic solution that contains only 300 mOsm/L of X (no Y is present in the external solution)?

Answer: The cell will swell until it bursts. Since X is permeable across the cell membrane, the only way that a stable steady state can be achieved is if [X]i = [X]e. So X will begin to move into the cell. However, as it does so, the osmolarity of the internal (cell) solution will increase above that of the external solution, so water will follow. To have equilibrium for X requires that [X]i = [X]e, but osmotic balance requires that [X]i + [Y]i = [X]e. These requirements can only simultaneously occur if the cell could swell to infinite volume (thereby diluting [Y]i to zero). Of course what will happen is that the cell will burst. Here the intraand extracellular fluid are iso-osmotic but hypotonic.

A somewhat more realistic situation (but still over-simplified) can be illustrated by considering red blood cells in plasma and the effects of the injection of various solutions. To make things simple, we will assume that there is no mixing between the plasma and the rest of the extracellular fluid (this, of course, is not realistic). So let us consider an individual with a total blood volume 5 L, made up of 3 L of plasma and 2L of red blood cells (RBC).

Also, for simplicity, assume that the RBC contain only solutes that are impermeable across the membrane, and that their internal osmolarity is initially 300 mOsm/L. Initially the osmolarity of the plasma is also 300 mOsm/L.

Question: What will happen to RBC volume and osmolarity if 1 L of distilled water is mistakenly injected intravenously?

Answer: To answer this question you should remember that:

Concentration !

Amount Volume

The total amount of solute in the RBC is 2L x 300 mOsm/L = 600 mOsmoles. The total amount of solute originally in the plasma is 3L x 300 mOsm/L = 900 mOsmoles. Adding distilled water does not change the amount, only the volume. So the total amount of solute in both the RBC and plasma is 600 + 900 = 1500 mOsmoles. After the injection of distilled water the TOTAL blood volume increases to 6L. Remembering the equation above, the new concentration is easy to determine, namely:

Concentration = 1500 mOsmoles/6L = 250 mOsm/L

The new steady state RBC volume is also easy to determine since the amount of solute in the RBC has not changed (600 mOsmoles, since all solutes are impermeable), but the concentration has changed to 250 mOsm/L. So:

250 mOsm/L = 600 mOsmoles/New RBC Volume

So, New RBC Volume = 2.4 L.

Similar problems will be considered in the first Membrane Workshop. This level of understanding of cell volume regulation may seem trivial, but it is certainly important to everyday clinical practice.

MEMBRANE PHYSIOLOGY QUIZ: Note that this quiz is simply provided to you to help you to test your understanding of the content of each lecture. They are not substitutes for studying the lecture notes (including Objectives and Take Home Messages), Practice Exams or other material intended to guide your learning. These questions are not intended to reflect the style of level of difficulty of questions on the block exam. MULTIPLE CHOICE QUESTIONS: 1. a. b. c. d. e. 2. The lipid bilayer portion of the membrane: is impermeable to most polar molecules and ions is highly permeable to most polar molecules and ions is very impermeable to all solutes contains a hydrophilic core made up of the hydrocarbon tails of lipid molecules is the only site of water transport across the membrane A cell with an initial concentration of solute particles of 300 mOsm/L is placed in a large volume of solution with a concentration of solute particles of 200 mOsm/L. Assume that the cell membrane is impermeable to all solutes both within the cell and in the solution. Once steady state is reached, the new volume of the cell (relative to its initial volume) will be: 0.67 1.0 (i.e., unchanged) 1.5 2.0 3.0

a. b. c. d. e.

The following information applies to the next three problems (5, 6 & 7): A cell initally contains only impermeable solutes with a concentration of 300 mOsm/L. The cell membrane is impermeable to both Na+ and Cl- but is permeable to glycerol. The cell is placed in a very large volume of solution containing 150 mOsm/L of Na+, 150 mOsm/L of Cl- and 300 mOsm/L of glycerol (assume no changes in extracellular concentrations occur). 3. a. b. c. d. e. Once a steady state is achieved, relative to its initial volume, the new volume will be: 0.5 0.67 1.0 (i.e., unchanged) 1.5 2.0 6. a. b. c. d. e. 7. a. b. c. d. e. The concentration of glycerol inside the cell in the new steady state will be:

0 mOsm/L 100 mOsm/L 200 mOsm/L 300 mOsm/L 600 mOsm/L The final intracellular osmolarity of the cell will be: 300 mOsm/L 450 mOsm/L 600 mOsm/L 900 mOsm/L can not be determined without more information

8. a. b.

Which of the following are characteristics of carrier mediated transport and ionic channels: specificity saturation

c. d. e. 9.

competition all of the above none of the above Which of the following solutes have significant permeability through the lipid bilayer portion of the membrane (i.e., primarily cross the membrane by dissolving in the bilayer and diffusing across it): ions dissolved gas molecules protein molecules in the intracellular and extracellular fluid all of the above none of the above

a. b. c. d. e.

TRUE-FALSE QUESTIONS: 10. 11. 12. 13. 14. 15. 16. 17. Simple diffusion is the primary mechanism by which ions cross the cell membrane. Active transport directly or indirectly utilizes metabolic energy to move specific solutes against their electrochemical gradients. Secondary active transport directly involves the hydrolysis of ATP. Facilitated diffusion involves either channels or passive carriers. Ionic channels (when open) normally allow far greater fluxes of solutes than do carriers. Iso-osmotic solutions are always also iso-tonic. Protein mediated transport via carriers will eventually saturate if the concentration of transported solutes becomes high enough. All channels throughout the body that allow Na+ to cross the cell membrane are expressed by the same gene.

Answers: 1. a, 2. c, 3. b, 4. e, 5. c, 6. d, 7. c, 8. d, 9. b 10. F, 11. T, 12. F, 13. T, 14. T, 15. F, 16. T, 17. F