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Cells Pitch
(An Introduction to Cytology)
Cells are very special to us. Recall that you have to at least be a cell to be alive. This fact alone illustrates the importance of cells. But cells are more than importanttheyre interesting! Each of the cells in our body is like a little organism all its own. Some, like white blood cells, behave in a very independent fashion. Most cells are satisfied to work together with their neighboring cells to create the structures of our bodies. But whether the cell is a little stay at home like the cells in our skin, or whether they wander around the body in search of invaders, like our lymphocytes, cells have a complex structure and physiology that we are probably only beginning to unravel. The study of cells is called cytology. Whole courses and curricula are devoted to cytology, so I can only really introduce you to the subject in this class. After this lab, I hope you will develop a deep appreciation for cells as well as a better understanding of what they are. You will no doubt study them more in physiology. All physiology occurs at the cellular level. Note, too, that most cells begin as generalists, the so-called stem cells, and wind up as specialists. This process of cell specialization is called differentiation. Well talk more about differentiation more when we discuss tissues. Lets get back to why cells have distinct forms. Part of the form of the cell is determined by the external membrane that keeps the cell separated from its internal environment, as well as the number and location of organelles it possesses. Lets look at these two features of the cell a little deeper.

Cell membrane
Martini, pages 20-33 The cell membrane is composed of phospholipids, which are phosphorylated forms of triglycerides, a common form of fat. The phospholipids form a bi-layer. Before I go on, picture in your mind, an Oreo cookie, you know the chocolate cookie with the cream filling. The cellular membrane is built like an Oreo cookie. Imagine the chocolate cookies on the outside are the phosphate groups. The frosting on the inside is composed of the fatty acids. Come to think of it, a cell membrane may be built exactly like an Oreo cookie! But I digress. Dotted throughout the cell membrane are integral proteins, which penetrate all the way through the cell membrane, as well as peripheral proteins. The peripheral proteins on

Types of cells
There are over 200 types of cells in the human body and I dont expect you to learn them. I have given you a handout of line drawings from electron micrographs of various types of cells which should give you a good idea of just how diverse they are. We will see some of these cells again when we study tissues. Note the shape of these cells. Keep in mind that form fits function.

2 the interior lining of the cell membrane are usually enzymes or anchoring proteins. The external lining of the membrane contains proteins that are used for receptors and markers. In addition, the cell membrane contains a sugar coating called the glycocalyx. These sugars help identify cells and are import for intercellular recognition. substances, although the substance enters the cell following the principles of simple diffusion: it travels from a high concentration to a low concentration and does not require any extra energy to do so. Hence the name, facilitated diffusion. The term facilitated means to be helped. In this diffusion, the diffusing molecule is being helped by a protein. Osmosis is the movement of water in and out of a cell following the principles of diffusion, but it is a bit more specific than that. It is the diffusion of water across a semi-permeable membrane. Water travels in and out of cells freely (no one is quite sure how). Because cell membranes are permeable to water but not necessarily to the material dissolved within it, dissolved substances must pass through protein gates (facilitated diffusion). Sometimes these gates are closed. This means water can travel through a cell membrane and leave the dissolved substances behind. Such are the conditions of osmosis. It is therefore equally important to note that osmosis is the diffusion of water through a semipermeable membrane. If the membrane described above is permeable to the substance dissolved in water, the action of the flow of the solution would be simple diffusion, not osmosis. In osmosis, only the water moves. To recall the term, osmosis, think of H2Osmosis. Active transport occurs when substances travel from an area of low concentration to an area of high concentration. Yes, its the opposite of simple diffusion. This requires energy! ATP, the main energy storing molecule of the body, is our chief source of energy. Active transport then differs from the previously described mechanisms of

Mechanisms of Transport
Martini, page 34, Table 2.2 There are five basic ways that material enter or exit the cell. These are: simple diffusion, facilitated diffusion, osmosis, active transport and filtration. Although this is more physiology than anatomy, lets examine them briefly Simple Diffusion is the movement of particles from an area of greater concentration to an area of lesser concentration. Because small molecules and ions are in constant motion, no additional energy is required. Facilitated Diffusion is similar to simple diffusion, in that no extra energy is needed, but a protein is required. Gases and fats can enter or leave the cell anywhere they wish, but most substances must be taken into the cell via specialized channel, gated or carrier proteins. The substances that require facilitated diffusion include all non-fat nutrients and ions. The protein acts as a doorway for a specific type substance trying to enter the cell, much as a doorway in a room allows you to enter or exit. No one would expect you to walk through the wall at least not this early in the semester. The protein, then, facilitates the movement of

3 molecular movement in that it requires energy while the others do not. Remember, it takes little energy to disperse molecules, but lots of energy to bring them together. What takes more energy, dropping a pack of cards, or picking them up? Filtration. If you have ever made coffee in a filtered system you have practiced filtration. Here small particles are separated from large ones. In the coffee example, we can rely on gravity to pull the liquid through and leave the coffee grounds behind. In the human kidney, filtration requires blood pressure. The cellular filters of the kidney filters (the glomeruli) are designed to separate substances in the plasma of blood. Blood cells are kept in as well as the blood proteins, but water, and any substances that are dissolved within it, are allowed to pass through to create a filtrate that will ultimately become urine. Well discuss this process briefly when we study the urinary system. To summarize for now, the separation of materials via a filter is called filtration. Any other name just wouldnt do. There are three distinct types of endocytosis. These are phagocytosis, pinocytosis, and receptor-mediated endocytosis. Lets quickly distinguish between them. In phagocytosis, the cell takes in large particles, or even bacteria, attaches a lysosome to them, and then digests them. Many white blood cells dine on bacteria. The process involves the cell sending an extension of itself to wrap around and ultimate engulf its lunch. Yum! The cellular cytoplasmic extension is called a pseudopod and it eventually rejoins the cell membrane. A vacuole is formed in the process, and a lysosome is attached to the vacuole that digests the particle or prey. Amoebas, microscopic amorphous protozoans, behave much the same way. When it comes to cell hunting we havent evolved much past the amoeba. Perhaps this also explains our government. The term phagocytosis literally means cell eating. May your cells dine in style. In pinocytosis, fluids, not solids, are taken in by a funnel shaped invagination in the cell that resembles a shallow pore. The outer edges of the pore eventually come together forming a small vesicle via miniature pseudopods. Incidentally, pseudopod means false foot. The term pod shows up a lot in scientific jargon. The early scientists were into feet. Pseudo is also common, as are most things false. As with phagocytosis, a lysosome attaches to the vesicle, digesting the fluid within. Pinocytosis literally means cell drinking. Cheers! The last endocytotic process is a little more complicated, but we wont cover it in great detail. During receptor

In and Out
(see Gartner & Hiatt Graphics 1-3 &14; Martini, pages 32-33.) Cells take materials into themselves and also produce materials for export. When materials are sent from cells, the process is called exocytosis. The extra cellular material of connective tissues is a product of exocytosis, as is the serous fluid produced by serous membranes. When materials are taken into the cell, the process is called endocytosis. Exo means out; endo means within.

4 mediated endocytosis, the cell has specific receptor proteins arranged in a shallow pocket on the cellular membrane. The receptor proteins typically attract only one type of molecule. The attracted molecules are sometimes called ligands. Once the ligands attach to the receptors, the pocket is pulled inward to form a small vesicle. Once again a lysosome attaches to the vesicle and helps to remove the ligands, releasing them into the cytoplasm. The lysosome then detaches from the vesicle and the receptor rich vesicular membrane makes its way to the surface on the cell. Once it reaches the surface, the vesicle forms shallow pocket again. Its a little like a bubble rising to the surface, but never quite completing its journey. Once this pocket is formed, the receptors are ready for more ligands.

Cytology in a Flash (A Quick and Dirty Guide to Organelle Function)

Nucleus: Brain of the cell. Usually the largest organelle. Nuclear Envelope: Double membrane of the nucleus; separates it from the cytoplasm. Cytoplasm: Everything in the cell, except the nucleus. Nucleolus: Ribosomes synthesized here. Ribosome: Sites of protein synthesis Rough endoplasmic reticulum: Cellular highway for protein transport and alteration. Contains ribosomes. Cellular membranes are remade here. Smooth endoplasmic reticulum: cellular highway for lipid synthesis. ribosomes absent. Resembles a vesicle, but its not round. Multiple Jobs: example, detox unit. Golgi apparatus: Refinery of the cell. Receives partially completed materials from the endoplasmic reticula. Lysosome: Digestive unit of the cell. Peroxisome: Another detox unit of the cell. Looks like a vesicle. Gee thanks. I wont ask to you to identify these. Centriole: Non-membranous organizer of cilia and cell division. Cilia: hair-like structures that move extracellular material across the cells surface. Like an automatic brush. Microvilli: Folds within the cell membrane to increase its surface area. Vesicle: Storage bubbles within the cell. produced by the golgi apparatus and or endoplasmic reticula.

Martini, page 29, Table 2.1 Recall that organelles are the little organs of the cells. If you examine the line drawings in your handout, you will see that all organelles are not found in each cell type and that their number and location vary considerably as well. To comprehend the purpose of each cell, we must first understand the function of the organelles. See your handout. One thing to note is that some organelles are membranous, having a phospholipids bilayer exterior, while others are non-membranous and are composed largely of proteins. Because the membranes of organelles resemble the cell membrane itself, one membrane can literally blend into the other. This is particularly true of vesicles and vacuoles, which serve to replenish the cell membrane.

5 Cytoskeleton: protein scaffolding of the cell. Helps make its form. Mitochondrion: the power house of the cell. My favorite organelle! You may also refer to Chapter 2 in Martini and Chater1 in Gartner & Hiatt, as well as your cell handout. Have fun.

How to Multiply by Mitosis

(Martini, page 44, figure 2. 20)
Cellular division is a result of two processes, the division of the nucleus, called mitosis, and the division of the cytoplasm called cytokinesis. When a cell divides, it generally splits into two smaller cells of equal size. Because these cells are essentially half the size of the parent cell, it is usually important for the cell to grow before it divides again. The initial growth phase or gap phase is called G1. Here more organelles are generated, more cytoplasm is created, and the cell does whatever job it was destined to do. Once the cell has reached an appropriate size and age, synthesis of DNA takes place. This is the S subphase. In the S subphase, DNA is replicated, so that the cell contains twice the genetic material it had during G1. This process takes 6 to 8 hours to complete. That may seem fast, but on the cellular level, thats pretty slow. But then the cell has to be careful. The exact replication of genetic material is important because it ensures that when the cell divides again, a full compliment of DNA will go to each of the daughter cells without any surprises. A surprise in DNA is called a mutation. Special proof reading enzymes ensure that mutation rates are very low, so a cells destiny is essentially chiseled in stone. In the final gap subphase G2, the cell prepares for cellular division. This subphase usually lasts only a few hours. The centrioles will play a key role here, as they will provide the guiding structural frame work for the chromosomes to separate. Chromosomes are bundles of DNA and protein. (Well discuss this process a little later). The centrioles finish replication during the G2 phase, ensuring that the DNA will be distributed correctly. When G2 is finished, mitosis is ready to begin. Note that G1, S and G2 are all subphases during the non-dividing phase of the cells life called interphase. Most of the cells life is in interphase. We use to say interphase was the resting phase. Oh contraire! The cell is far from resting. During interphase, the cell grows, replicates its DNA, and yes, does whatever work it was meant to do. It should be called the working phase. Im not finished complaining about interphase. I have another gripe about this term. Inter means between, and one would guess that it means it is the phase that lies between mitotic divisions. Thats fine, but not all cells continue to divide. Skeletal and cardiac muscle cells dont divide, neurons dont divide, in fact most differentiated cells dont divide, and they encompass nearly all of our cells. Cells that can no longer divide are in the G0 phase. This is not an in between phase; this is an end-of-the-

6 line phase. Nonetheless, we are forced to say that cells in G0, are in interphase just to avoid non-conformity. Like I care about that! OK! OK! Dont be impatient! Mitosis is just around the corner. But first a word about chromosomes. Its an important digression, I assure you. (See Martini, page 39, figure 2.13). Chromosomes (meaning colored bodies, simply because they stain well) are visible only during mitosis. Prior to mitosis, the DNA is in the form of chromatin. Chromatin consists of loosely coiled threads of DNA that wrap around bundles of protein, called histones, like stripes on a barber pole. The histones are usually depicted as little spheres lumped together in clusters of eight. The clusters are separated from each other by special strips of DNA called linker DNA. The combination of DNA and histone clusters forms the fundamental unit of chromatin called the nucleosome. When a cell is preparing for mitosis, the histones of the nucleosomes begin to separate and each strand of DNA becomes more tightly bound to each individual histone, as opposed to the whole histone cluster. Each histone then resembles a doubled striped ball, with DNA wrapping around it. (Note that the figure in Martini doesnt do this relationship justice, but its a minor point.) The linker DNA also becomes shorter, forming a tightly coiled structure. As the structure gets tighter, it also becomes thicker and eventually it becomes visible. Once it becomes visible, the DNA/histone structure is called a chromosome. Because DNA was replicated in the synthesis (S) phase, there are two coils of DNA per chromosome. We refer to each of these coils as chromatids. (Two chromatids make one chromosome). Whats interesting about the chromatids in the same chromosome is that they are genetically identical. If youre wondering why that is, refer to Martini, pages 4445, figure 2.21. The two chromatids are pinned together in a special region called a centromere by specific proteins called kinetochores. The specific name for these proteins is cohesions. If youre ever on Jeopardy, youll need to know this. The centromere is a key feature in successful mitotic division and will be discussed later. Frankly, Ill be delighted if you remember the term centromere; the rest can wait for a cytology class.

Mitosis or Bust!
(Martini, pages 46-47, figure 2.22; Gartner & Hiatt, p. 17, plate 1.4)
Heres a brief overview of mitosis. Mitosis comes from the Greek word mitos, which means threads. During mitosis several things happen:

7 1. 2. 3. The chromosomes become visible, 2. The nucleus and nucleoli disappear, The chromosomes resemble short fat threads as they line up centrally within the cell. The chromosomes pull apart at their centromeres, and The halved chromosomes migrate to opposite poles. As new cells form, two new nuclear membranes form around the chromosomes Finally cytokinesis, the division of cytoplasm, typically follows. but you cant see everything thats happening. An electron microscope could reveal the centrioles that are migrating to opposite ends (poles) of the cell. The centrioles create spindle fibers that attach at the centromeres, which are tubular rods of proteins that will be used to move the chromosomes into a position that will ultimately pull them apart. Some spindle fibers also bypass the centromere to butt up against spindles coming from the opposite centrioles. These spindle fibers form a framework for chromosomal division. This framework is a diamond shaped spindle apparatus. Under the compound microscope, the spindle apparatus will become obvious to us in the next phase, metaphase. Metaphase is the second phase of mitosis. During metaphase the chromosomes line up at the center of the cell (in most cases). Their chromatin arms often project from the line resembling locked fingers. The spindle apparatus, as promised, is also visible. The chromosomes are still in tact, waiting for separation. We are in the middle of mitosis now. Met means middle. Anaphase is the phase of chromosome separation. Here the chromosomes separate into chromatids at the centromere. The chromatids then pull apart and the combination of chromatids and spindle apparatus resembles two umbrellas, one pointing up, and one upside-down. Indeed the prefix ana means up. I guess it was too much to include down in the terminology. (Please note that Im using the term chromatid here to avoid some confusion. Ill talk about the real name of the divided chromosome later).

4. 5. 6.


Thats the short version of cellular division. Now lets examine the specific phases in a little more detail. Prophase is the first phase of mitosis. (Pro means before; we are definitely in a state of preparation). During interphase, a stained cell displays an obvious nucleus with nucleoli. During prophase, the nucleoli disappear and the colored mass of the nucleus becomes darkly marbled. As prophase progresses, the nuclear envelope disappears. The chromosomes become more and more obvious until they resemble noodles. The transition of chromatin to chromosome is equivalent to going from soup to noodles! You can easily recognize prophase using a compound microscope,

8 Chromosomes soon appear, displaying both sets of DNA, one for each chromatid and two chromatids make the chromosome. Now comes the tricky bit. When the chromosome separates during anaphase, the chromatids are given a different name. They are now called daughter chromosomes. In a sense, the chromatids have graduated to the state of chromosome because it is now up to each of them to carry on the genetic message alone. As daughter chromosomes complete their migration at the end of telophase, they will give each of the two new cells one set of DNA, not two. If either of these cells is destined to divide, it will eventually progress to the S subphase, at which time the cell will create a complimentary set of DNA. When its chromosomes finally appear in prophase, each chromosome will be composed of two chromatids with identical strands of DNA. But we only need one, right? The other is just a copy, but a necessary copy when this cell divides. Duplicating the DNA in this way ensures that the two new daughter cells will be genetically identical, not only to themselves, but to the mother cell that produced them. The process repeats itself again and again in our lifetimes, depending upon the type of cell. One last thing. It is important to realize that mitosis is a fluent action. The transition between phases is gradual, not abrupt, and many intermediate phases exist. You will often have to make a decision as to which phase the cell is in. Im not going go to get too excited about this as long as you clarify your answer. For example, if you see a cell somewhere between anaphase and telophase, you could call it late anaphase or early telophase. Lets not split hairs over this. I only ask that you know enough to make

Telophase is the final phase of mitosis. We know were in telophase because it marks the end of chromosomal migration. Sorry, we cant see this migration. Everything is locked dead in its tracks, so to speak. Still, we see the chromosomal clusters that look like squashed bugs at opposite ends of the cell. Sometimes, a cleavage line can be seen between them, showing the beginnings of cytoplasmic division (cytokinesis). Then it appears that we are looking at not one cell, but two. As telophase progresses, the nuclear envelope reforms, the spindle apparatus vanishes, and eventually nucleoli reappear. Needless to say the wad of chromosomes starts to uncoil into chromatin, a transition from noodles back to soup. The prefix telo- means end. Well, thats the end of mitosis, but not the end of cellular division. Before I discuss cytokinesis, we need to resolve the matter of the chromosome. Let me preface my remarks by saying that there were no evil biologists trying to make learning cytological terminology difficult for students. It just happened that way. What I am about to tell you may sound a bit confusing, but Ill do my best to give you a satisfactory explanation. To understand the real terminology, it is important to review what weve covered so far. Remember Chromatin? Chromatin is the soupy state of DNA and histone protein that we find during interphase, but lets examine it more closely at the G1, S and G2 subphases of interphase. During G1, we have one set of DNA. Synthesis comes along, replicating the DNA, so that in G2 we have two sets of DNA. Clear so far?

9 an educated guess. During your examination, I will try to avoid the intermediate phases, but its fun to try to spot them. Ready for cytokinesis? The usual finale for cellular division is cytokinesis. Here the cytoplasm separates so that two distinct cell membranes form creating two cells called daughter cells. As you might guess, this process is relatively complex and best left for a cytology class. Still it is necessary to understand a few facts about cytokinesis. Cytokinesis is not requisite for all cells. Some nuclei divide without cytokinesis. The megakaryocytes that produce blood platelets, and the osteoclast cells that are responsible for tearing down bone, undergo further mitotic divisions without forming daughter cells, resulting in large multinucleated cells. Remember, these cells are the exception, and not the rule. If it were not for cytokinesis, we would be one amorphorous mass. Cellular differentiation would be impossible. We would have no neurons, no muscles, no glands, no skin. While mitosis, nuclear division, is important for the transfer and preservation of genetic material, cytokinesis ultimately sets the stage for cellular specialization.

Heres a little way to remember the phases of interphase + mitosis I = interphase Passed = prophase My = metaphase Anatomy = anaphase Test = telophase

Mitosis Hand Theatre Heres one more way to remember the phases of mitosis. You can use your hands to replicate the interphase and mitosis. Heres how. 1. Interphase: Fold your hands and lock your fingers so your hand resembles a ball. 2. Prophase: Wiggle your fingers. Your fingers represent chromosomes. 3. Metaphase: Start to pull your hands apart, but leave them locked in place.

10 4. Anaphase: Pull your hands apart. 5. Telophase: Make a fist with each hand. 6. Once you have achieved the two-fisted state, your fists represent two new cells, the daughter cells. 7. Practice this often, but dont do this in public unless you are not afraid to look like a lunatic. 8. Be happy.

Draw the phases of Mitosis and win a Buick!*






*Void in California