E X P E C TAT I O N S Explain the dynamics of the transport of substances through the cell membrane, including facilitated diffusion. Design and carry out an investigation to examine the movement of substances across a membrane. Figure 1.30 Two different water environments meet when the Thompson River joins the Fraser River, much like the internal environment of a cell and the extracellular fluid meeting at the cell membrane. The conditions inside every cell must remain nearly constant for it to continue performing its life functions. The steady state that results from maintaining near-constant conditions in the internal environment of a living thing is called homeostasis. The structure chiefly responsible for maintaining homeostasis inside a living cell is the cell membrane. You have seen that the cell membrane’s structure is remarkably complex. The cell membrane uses several methods to transport molecules of different sizes and with different properties in and out of the cell. The primary methods it uses rely on the fact that the cell membrane is semi-permeable, allowing some molecules to pass through it while preventing others from doing so. This section will examine those transport methods that involve substances moving through the cell membrane. On both sides of the cell membrane, water is the solvent, the meeting place for all of the other chemicals. As you learned in Section 1.1, water has special properties that make it a functional medium for living reactions. For example, the external environment of a single-celled life form, such as the amoeba shown in Figure 1.31, consists primarily of water. This external environment also contains other microscopic aquatic organisms, decaying organic matter, and dissolved gases (such as oxygen) and other inorganic substances. Figure 1.31 This amoeba has little sensory equipment, limited locomotion, and a seemingly fragile membranous covering. Yet it copes with an external environment as complex as yours. In the case of a multicellular organism, every cell is bathed in a thin layer of extracellular fluid. The extracellular fluid consists of a variable mixture of water and dissolved materials. Some are substances that a particular cell type requires; some are substances needed by all cells. Other materials are wastes that the cell has already discarded — and that the organism will eventually get rid of. Diffusion and the Cell Membrane One passive method by which small molecules move through the cell membrane is diffusion. Diffusion is the movement of molecules from a region where they are more concentrated to one Exploring the Micro-universe of the Cell • MHR 25 where they are less concentrated. Many molecules — especially small, uncharged ones, such as oxygen — can move easily through the cell membrane as a result of this process. How does diffusion work? You may remember from earlier studies that molecules are in constant motion — even in a solid. In a liquid, this means that the molecules move about randomly all the time. As they collide with each other and with the walls of their container, they rebound, changing speed and direction. This constant, random movement of molecules in a liquid is called Brownian motion. It drives the process of diffusion. If molecules of another substance are added to water, they will be bounced around by the motion of the water molecules and each other until the new substance is spread evenly throughout the water. From earlier studies, you will recall that in this case the water is acting as a solvent that dissolves other substances, or solutes. Diffusion always results in the net movement of particles from a region of high concentration toward a region of low concentration. The difference in concentrations between these regions is called the concentration gradient. For example, in the river pictured in Figure 1.30 on the previous page, the concentration of mud particles is very high on one side and very low on the other. What do you think happens to the concentration of the mud particles farther downstream? Over short distances, diffusion works well to transport small molecules across the cell membrane. For example, oxygen and carbon dioxide cross the cell membrane by diffusion. As a cell uses up dissolved oxygen, more oxygen enters the cell; as a cell generates carbon dioxide, more carbon dioxide leaves the cell. Diffusion Limits Cell Size Diffusion also explains how molecules move around once inside the cell. But the concentration gradient within a cell is not nearly as great as that across the cell membrane. Once molecules have diffused through the membrane, their rate of diffusion slows down abruptly. How does this fact limit the maximum size of cells? Figure 1.32 gives some clues. For the cell, having a large surface area relative to its volume increases the area available for materials to diffuse in and out. Random Walking Analyze In this lab, you will measure how long it takes for foodcolouring particles to diffuse different distances. Fill a 25 mL graduated cylinder with warm tap water. Gently tap the side of the cylinder to eliminate all air bubbles in the water. Use a long pipette to take a 1 mL sample of undiluted blue or red food colouring, and then rinse the outside of the pipette with running water. Carefully insert the pipette into the cylinder until the tip reaches the bottom of the water. Release the food colouring into the water. Time how long it takes for the colour to move 3 mL up the cylinder. Time how long it takes for the colour to move up 10 mL. 26 MHR • Cellular Functions Figure 1.32 This amoeba has two problems: (1) It would take years for molecules critical to survival, such as oxygen, to reach its centre via diffusion. (2) Relative to the volume of the “body” that it has, it does not have much surface area (cell membrane) across which substances can move in and out. 1. Make a general statement about the speed of diffusion and distance. 2. Explain why you think this MiniLab is called “Random Walking.” 3. Make a prediction about what would happen to the rate of diffusion if you (a) increased the temperature of the water or (b) decreased the temperature of the water. In each case, explain your reasons why. 4. State whether temperature is a dependent or an independent variable in testing rates of diffusion. As a cell increases in size, what happens to the amount of surface area it has relative to the volume of its contents? Find out by calculating the ratio of a cell’s surface area to its volume. Start your calculations using a cube-shaped cell with sides 1 unit long. Find the cell’s surface area and volume. Divide the surface area by the volume. Increase the length of each side by 0.5 units, and perform your calculations again. Repeat this 10 times. Display your data in a spreadsheet. Perform the same calculations for a spherical and a cylindrical cell (both starting with a radius of 1 unit). Display your data in a spreadsheet. How does changing the shape of the cell affect the ratio of surface area to volume? Osmosis: The Diffusion of Water Water from the extracellular fluid and from inside the cell also diffuses freely through the cell membrane in such a way that the concentration of water on either side of the membrane usually remains equal. This diffusion of the solvent across a semi-permeable membrane separating two solutions is called osmosis. For cells, where the solvent is water, the water molecules move from a region of higher concentration to a region of lower concentration — as in any other form of diffusion. The direction of osmosis always depends on the relative concentration of water molecules on either side of the cell membrane: hypotonic environment. The figure also shows how the concentration of solutes on either side of the cell membrane affects the concentration of water. One of the dangers a cell faces under hypotonic conditions is that the cell membrane may burst. The destruction of a cell through this process is called lysis. What would happen to this cell under hypertonic conditions? When water diffuses out of the cell, the process is called plasmolysis. A higher concentration of solutes in the extracellular fluid than in the intracellular fluid can cause plasmolysis. Equal water concentrations Greater water concentration outside cell — pure water When the water concentration inside the cell equals the water concentration outside the cell, equal amounts of water move in and out of the cell (isotonic conditions). When the water concentration outside the cell is greater than that inside the cell, water moves into the cell (hypotonic conditions). When the water concentration inside the cell is greater than that outside the cell, water moves out of the cell (hypertonic conditions). The cell membrane cannot prevent this movement of water (because it is permeable to water molecules), and the only energy involved is the Brownian motion of the water molecules. Hence, osmosis (like diffusion) is a passive process that does not require energy from the cell. Clearly though, the cell can remain healthy only if the water concentrations inside it and in the extra cellular fluid surrounding it stay in balance. Blood plasma and the fluid that bathes our cells are usually isotonic. Figure 1.33 illustrates what happens to a red blood cell in an isotonic and a Figure 1.33 If a cell is placed in a hypotonic solution, water enters the cell by osmosis. Under these conditions, cells without cell walls may burst. Facilitated Diffusion Although water, oxygen, and carbon dioxide can pass through the cell membrane without assistance, other substances cannot do so without help. This makes the cell membrane a selectively permeable membrane. For example, glucose cannot cross the cell membrane by simple diffusion — even if the glucose concentration outside a cell is greater than that within. The glucose molecule is too big to diffuse between the phospholipid molecules of the Exploring the Micro-universe of the Cell • MHR 27 membrane and is insoluble in lipids, so it cannot dissolve in the lipid bilayer. How then do molecules such as glucose get in and out of the cell? This is where many of the proteins studded in the cell membrane play a role. Specialized transport proteins in the cell membrane help different kinds of substances move in and out of the cell. The structure of these transport proteins enables them to be highly selective. A particular transport protein will recognize and help to move only one type of dissolved molecule or ion based on the particle’s shape, size, and electrical charge. In the case of glucose, a type of membrane protein called a carrier protein facilitates the movement of glucose molecules from a region where they are more concentrated to a region where they are less concentrated. Because the relative concentration of glucose still drives its movement through the carrier protein, this facilitated diffusion is also an example of passive transport. Figure 1.34 shows how a carrier protein works. Osmosis in a Model Cell The small size of living cells makes it difficult to observe osmosis actually occurring across their membranes. However, you can make a model of a cell to study the process of osmosis. You can use dialysis tubing, which provides a synthetic membrane permeable to water molecules, as the membrane of a model cell. In this investigation, you will design and conduct an experiment to determine how the composition of the extracellular fluid affects osmosis. Problem Hypothesis How does the presence or absence of solutes in the extracellular fluid affect the direction and amount of osmosis through the model cell’s membrane? Make a hypothesis about the effect of solutes in the extracellular fluid on the flow of water through the cell membrane by osmosis. CAUTION: Follow your teacher’s directions for conducting laboratory experiments safely. Do not consume any food products in the laboratory. Materials dialysis tubing string to tie tubing support stand to hang “cell” in beaker large beakers (500 mL) or jars distilled water, egg white, or Ringer’s solution 4 mol/L sucrose solution or table syrup sodium chloride (table salt) tap water any other materials you require Experimental Plan 1. Examine the materials provided by your teacher. As a group, list the possible ways 28 MHR • Cellular Functions Carrier proteins depend on their threedimensional shapes to do their jobs. A carrier protein will accept only a non-charged molecule with a specific shape, much as a shape-sorting toy allows a child to insert a triangular piece only in a triangular hole. However, carrier proteins allow molecules to move both in and out of the cell. To review cell membrane structure and enhance your learning about membrane transport, go to your Electronic Learning Partner. Figure 1.34 Carrier proteins change shape to allow certain molecules to cross the cell membrane. you might test your hypothesis using these materials. 2. Agree on one way that your group could investigate your hypothesis. 3. Your experimental design should use a control and test only one variable at a time. Plan how you will collect quantitative data. 4. Write a numbered procedure for your experiment that lists each step, and prepare a list of materials that includes the quantities you will need. Checking the Plan 1. What will be your independent variable? What will be your dependent variable(s)? How will you set up your control? 2. What measurements will you make? How will you determine if the solute content in the extracellular fluid has affected the direction or amount of osmosis? Have you designed a table for collecting data? 3. How many trials will you carry out? How long will you allow each trial to run? Data and Observations Conduct your experiment, make your measurements, and complete your data table. Design and complete a graph or other visual presentation of your results. Analyze 1. How did any changes you observed in your model cell(s) relate to the solute concentrations of the extracellular fluid in which you placed the cell(s)? 2. What evidence do you have that the amount of water inside the cell was or was not changed by osmosis? 3. What evidence do you have that dialysis tubing is permeable to water? Conclude and Apply 4. How do solutes affect the concentration of water? 5. Based on your results, predict how the use of road salt to melt ice or snow affects the plants bordering the sidewalk, road, or highway. Explain. Exploring Further 6. Using what you learned in this investigation, design an experiment that would help solve a related problem: How will the presence or absence of solutes in the model cell’s intracellular fluid affect the direction and extent of osmosis through the model cell’s membrane? If time and materials are available, perform the experiment and anlayze your results. Exploring the Micro-universe of the Cell • MHR 29 No cellular energy is required regardless of whether the substances move in or out of the cell. Facilitated diffusion does require the participation of specialized membrane proteins, but the proteins do not require energy from the cell to do their job. Active Transport Figure 1.35 Channel proteins provide water-filled passages through which small dissolved ions can diffuse. Channel Proteins Since carrier proteins cannot transport charged particles across the cell membrane, a different type of membrane protein called a channel protein does this. Channel proteins have a tunnel-like shape that allows charged particles (ions) to pass through the lipid bilayer. Figure 1.35 illustrates how this process works. To pass through a channel protein, an ion in solution must be small enough to fit through the “tunnel.” It must also have the right charge. In much the same way that like poles of two magnets repel each other, a positively charged channel protein repels positively charged ions and a negatively charged channel protein repels negatively charged ions. A cell often needs to maintain an intracellular environment vastly different from that outside the cell. For example, it must concentrate nutrients for growth and maintenance inside, and also carry out any specialized functions it may have. In addition, many of the cell’s waste products are highly toxic and must be completely removed from the intracellular environment. Passive transport would allow some of these materials to remain in the cell. How does the cell acquire control over the substances it needs for life? To do this, the cell must expend energy to transport substances (solutes) from an area of lower concentration to one of higher concentration, much like pushing an object up a hill. This process of moving substances against (or up) their concentration gradients is called active transport. How much energy does it take to move a substance up a concentration gradient? That depends on how steep the uphill gradient is. Particles move down a concentration gradient with the same ease that you might ride a bike down a hill. A similar analogy can be made for particles being moved up a concentration gradient. Like the cyclist in Figure 1.36, the steeper the hill the harder the you have to pedal to get up it. In cystic fibrosis, a genetic disorder, faulty channel proteins cause chloride ions to build up outside the cell and sodium ions to build up inside the cell. This makes water move into the cell (by osmosis). The cells that line the lungs, intestines, and pancreas take water from the mucous layer coating the passageways, leaving the mucous thick and sticky. In the lungs, the thick mucous interferes with breathing. It interferes with the absorption of nutrients in the intestine. Intensive research has led to an increase in the quality and length of life for people living with the disease. In all of the forms of passive transport you have looked at, any substances crossing the cell membrane travel along (or down) their concentration gradient. 30 MHR • Cellular Functions Figure 1.36 Like a cyclist pedalling up a steep hill, the cell must expend a great deal of energy to pump molecules and ions in or out of the cell against their concentration gradients. for drinking or to purify municipal and industrial wastewater before discharge to the environment. ZeeWeed is composed of thin, hollow fibres. The membrane of the fibres has pores small enough to block the passage of viruses and of micro-organisms such as bacteria. These fibres are mounted in an open frame that can be immersed directly in the water to be treated. Like living seaweed, ZeeWeed fibres float freely. ZeeWeed has low energy requirements. A light stream of air bubbles keeps the Zeeweed fibres moving, thereby exposing the fibre membranes to incoming water currents. A slight suction on the clean water side draws water through the pores of the membranes into the hollow interior of the fibres, leaving the micro-organisms and viruses behind. Dr. Pierre Côté with a sample of ZeeWeed membrane and a glass of water that has passed through the membrane. Civil Engineer Traditionally, “membrane” is used to refer to a thin film with microscopic pores that admit only particles small enough to pass through. Membrane filtration technology beyond teabags and coffee filters is nothing new. For example, hospital dialysis machines use membranes to filter the blood of patients whose kidneys no longer function. However, the use of this technology is costly. Until recently, membrane technology for water filtration has been costly, too. Enter Dr. Pierre Côté, civil engineer, with a new kind of membrane. A book about the potential environmental impact of Earth’s rapidly growing human population motivated him to focus on environmental engineering, especially water treatment. A typical water treatment plant passes water through clean sand as a primary filter and then gel-like coagulants to trap fine sediments, such as clay. After this, chlorine is added to kill remaining bacteria. (A conventional “activated sludge” sewage treatment plant uses bacteria to break down wastes.) Now, Dr. Côté’s prize-winning membrane technology offers a new approach. Designing an Award-Winning Membrane Dr. Côté received a Manning Innovation Award of $100,000 for his development of ZeeWeed, a unique filtration membrane that represents a revolution in water treatment. It can be used to treat ground or surface water Dr. James M. Dickson of McMaster University’s Membrane Research Group describes such membranes as “not very smart” when compared with the cell membrane. “They’re designed to do a very specific job, and that’s all they do.” The membrane around a living cell must sense and respond to every aspect of the internal and external environment. It must perform dozens of distinctly different sensing, separation, and transportation tasks — some of them thousands of times per minute. Teamwork Pays Off As the chief technical officer at Oakville’s Zenon Environmental Inc. — a global leader in advanced membrane technologies for water purification, wastewater treatment, and water recycling — Dr. Côté loves his job. “Just coming to work is fun. It’s not working,” he says. “I’m doing research and development, so it’s always something new.” When asked what he’d do with the prize money from the Manning Award, he replied “I will share it with my [research] team and the workers at Zenon.” Like other researchers in both the pure sciences and the technologies, Dr. Côté recognizes that new developments almost always result from a team effort. Career Tips 1. Research further to discover how civil engineers are solving other real-world environmental problems. 2. What knowledge of cell biology should a manager at a water-treatment facility have to do the job effectively? How does this person work with the Ministry of the Environment, water-testing facilities, and local landowners to ensure the delivery of safe drinking water? Exploring the Micro-universe of the Cell • MHR 31 When a person is resting, his or her cells use up to 40% of their energy on active transport. Many types of specialized cells use much more. For example, the cells in your kidneys that filter your blood use up to 90% of their energy on active transport. What kind of substances do cells need to pump in or out by active transport? A few examples follow: Kidney cells pump glucose and amino acids out of the urine and back into the blood. Intestinal cells pump in nutrients from the gut. Root cells pump in nutrients from the soil. Gill cells in fish pump out sodium ions (their extracellular fluid is less salty than sea water). Figure 1.37 How does the work done by a refrigerator resemble the work done by active transport in cells? Specialized cells in your stomach lining secrete acid so that you can digest foods. They maintain the concentration of acid at about 0.000 15 g/mL of fluid. That number may seem low, but the concentration of acid inside the cells is much lower — less than 0.000 000 000 05 g/mL. Compare the acidity of the fluid inside your stomach to the acidity inside these cells: divide the stomach’s acid concentration by the cell’s acid concentration. Write your answer using scientific notation. What does this ratio tell you about the concentration gradient faced by the cells? Why must the cells consume large amounts of energy to pump acid out across their cell membranes? It may help to think of the cell as a refrigerator. Although food comes into the kitchen, some food must be actively concentrated in the refrigerator and some items in the refrigerator need to be taken out. The refrigerator has to maintain a special environment inside in order to keep the food fresh. To do this, the refrigerator uses a mechanism to cool the inside air and pump out excess heat. It also removes the water from the air inside and sends it outside (the water comes from the food that enters the refrigerator). Otherwise, the water would condense on surfaces in the cold Relative Concentration Challenge Nitella can spread across the bottom of a pond, providing refuge for many pond dwellers. Pond water contains several of the dissolved ions needed by pond organisms, such as sodium (), potassium (), calcium (), magnesium (), and chloride (). In this lab, you will investigate the concentration of these ions inside and outside of Nitella. The vertical blue bars on the graph represent ion concentrations inside the Nitella cells. The green bars represent the concentration of ions in the pond water outside Nitella’s cell membrane. You Try It Background Nitella, a green alga, is a common pond organism that has very long cells and a plantlike structure. If left undisturbed, 32 MHR • Cellular Functions 1. If ions simply diffuse through Nitella’s cell membrane, how would the blue and green bars for each ion compare? Suggest why simple diffusion cannot account for the data presented in the graph. 2. What evidence is there that Nitella must somehow be forcing ions inward against a concentration gradient? environment. Every time you open the refrigerator, warm air flows in and cold air flows out, both along their concentration gradients. Even with the door closed, the refrigerator pump must still come on frequently and use energy to maintain the special internal environment. Active Transport Pump The cell uses an elegant system to actively transport substances in and out against their concentration gradients. The engine that drives this system is a pump, which runs on energy from cell metabolism. The pump is a cell membrane protein. This transporter protein actively pumps ions across the membrane against their concentration gradients. Cells have several different transporter pumps. The best-understood example of an active transport pump is the sodium-potassium pump in animal cells. The cell membrane of every cell in your body uses these pumps. As Figure 1.38 shows, this transporter pumps sodium and potassium ions. When three (positive) sodium ions inside the cell and two (positive) potassium ions from the extracellular fluid bind to the transporter’s protein complex, the transporter taps a form of cellular energy (ATP). This allows the protein to change shape. In its new shape, the three sodium ions move to the outside of the cell and the two potassium ions move inside — the transporter has flipped the ions. Then it releases all of the ions and returns to its original shape. To learn more about cellular respiration and ATP, turn to Chapter 3, Section 3.3. To view an animation of and explore active transport, go to your Electronic Learning Partner. Tapping the Energy Stored by Active Transport The cell uses the artificial concentration gradient it has created for sodium ions to push molecules it needs, such as glucose and amino acids, into the cell. The cell cannot function if it only gets as many of these molecules as diffusion will allow into the cell, so the cell must move extra glucose and amino acids in against a concentration gradient. Figure 1.38 The primary components of the activetransport system driven by the sodium-potassium pump How can a sodium ion exert a pushing force on a molecule? Think of the sodium ions as skiers at the top of a ski hill with nowhere to go but down. The cell pushed them up the chair lift. Now it makes them take a molecule down the hill with them. A type of carrier membrane protein helps the sodium ion and a molecule (such as glucose) enter the cell. When one sodium ion and one glucose molecule bind to this carrier protein, it changes shape. It now allows the sodium ion to ride down its concentration gradient into the cell — providing the energy to move the glucose molecule as well. Plant and bacterial cells use hydrogen instead of sodium ions to do this. Another protein in the cell membrane also taps the energy stored in the sodium-ion concentration gradient — this time to push another positive ion out of the cell. A common use for this exchange of one ion for another is pumping unwanted hydrogen ions () out of the cell against their concentration gradient. This keeps the cell interior from becoming too acidic. The artificial concentration gradients that active transport creates for sodium and potassium ions result in a constant tendency for potassium ions to diffuse out of the cell and for sodium ions to diffuse back into it. So the sodium-potassium pump must work constantly. In fact, even when you are resting it consumes nearly one third of the energy generated by your cells. This high-energy requirement is thought to be caused by the need for Exploring the Micro-universe of the Cell • MHR 33 rapid, repeated changes of shape in the transporter protein complex. Through its active transport system, the cell stockpiles nutrients it needs for maintenance and growth and pumps out unwanted particles. In addition, it creates an electrical potential across the cell membrane that allows nerves and muscles to work. The higher concentration of positive ions outside the cell creates an electrical charge across the cell membrane. 1. 34 Define diffusion using one specific example. 2. Explain the concept of a concentration gradient, and use diagrams to clarify your explanation. 3. Identify three different molecules that diffuse into cells. 4. Distinguish between osmosis and diffusion. 5. What is homeostasis? Why is homeostasis important to cells? 6. Diffusion allows for the effective movement of substances over short distances. How is this important for the cell? 7. Some potato cells are immersed in tap water. Make diagrams showing the relative concentration of water in both the cells and in the water. 8. Describe the movement of water inside a cell in a hypertonic environment. 9. How is facilitated diffusion different from diffusion? For a nerve impulse to travel the length of a nerve cell, it must keep changing the electrical charge across the cell membrane as it goes. The change in charge in one part of the nerve cell membrane causes special ion channels to open in the next part. During the millisecond that each of these sodium ion channels remains open, some 7000 sodium ions pass into the cell through each one. To allow for a new nerve impulse, the cell must pump out all of these sodium ions. 15. Cystic fibrosis is the result of genetic mutations within a gene identified as cystic fibrosis transmembrane conductance regulator (CFTR), which codes for a channel protein. An experiment was conducted to identify the specific ion being transported by CFTR. The candidate ions were chloride (), magnesium (), and sodium (). The experiment involved comparing cytoplasmic levels of these ions between normal cells and cystic fibrosis cells. Interpret the results below, and give a possible explanation for them. 16. Plan a dramatic presentation involving students in the class to show the difference between passive transport and active transport in the cell. 10. Identify two situations where cells need to use active transport. Describe an additional possible situation where active transport might make sense. 11. Make a diagram of the sodium-potassium pump, and briefly explain how it works. Cells need to bring in (absorb) nutrients. • What method(s) of transport do cells in the intestine use to absorb nutrients? Explain why. 12. What would happen to a cell if its cell membrane were permeable rather than semi-permeable? • Draw a picture showing how glucose and galactose enter a cell. Use a different shape for each simple sugar. 13. Why does oxygen continue to diffuse into cells on an ongoing basis? 14. Explain why water pollution has such a profound effect on living organisms. • If a cell has adequate glucose in its extracellular environment, explain why a cell might not have enough glucose inside. Give several possible reasons. MHR • Cellular Functions