Cell Division. Learning Objectives: Introduction. Revised Fall 2018
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1 Revised Fall 2018 Cell Division Learning Objectives: 1. Define cell cycle and the ordered sequence of events in the cell cycle (Interphase and The divisional phase or M phase) 2. Explain the stages in interphase (G 1, S and G 2) 3. Understand the separation of chromosomes during mitosis. 4. Know the major events during cell division and their sequence (Prophase, Metaphase, Anaphase, Telophase and Cytokinesis) 5. Recognize mitotic phases in both plant and animal cells. Introduction Multicellular organisms generally begin life as a single fertilized cell, the zygote. These organisms must then repeatedly copy this cell to produce the many cells needed to make up the organism. The process used for this event is referred to as the cell cycle. The Mitotic Phase is that part of the cell cycle where a single cell divides its chromosomes (mitosis) and its cytosol (cytokinesis) to produce two identical daughter cells. It is also used for growth, maintenance and repair of tissues in multicellular organisms. You will be observing interphase, and the stages of the mitotic phase: prophase, prometaphase, metaphase, anaphase, telophase and cytokinesis in both plant and animal cells. Interphase and the Mitotic phase make up one cell cycle. Fig. 1 Summary Diagram of the Cell Cycle. G1 or G0, S and G2 make up Interphase. Prophase, Prometaphase, Metaphase, Anaphase, Telophase and Cytokinesis make up the Mitotic (M) phase. Be sure and mark Interphase and M phase on this diagram. Note: What you will be studying in lab today is mitosis which you can see in the diagram is just part of the cell cycle. The S phase is when the DNA is replicated to form the sister chromatids. You will be following their movement during mitosis. 1
2 Activity 1 Mitosis in Plants A) To observe stages of plant mitosis use the slide in your slide box labeled Onion or Allium root tip that contains 3 cross sections. Using the low power of your microscope examine one of the root tips noting the number of cells that have darkly stained filamentous chromosomes. Now increase the magnification up to 400X and using the mechanical stage move the slide to find each of the mitotic stages described below. You may need to look at all 3 root tips to find all of the stages. As you locate each stage make a drawing in the space provided and be sure to label your drawing. Figures 2A-2F are model representations of the various stages of mitosis that you should observe. Interphase (Figure 2A). As noted in Fig. 1 Interphase is the non-dividing stage of the cell cycle. During this time the nucleus is visible and contains 1 or more small, round, darker-staining structures called nucleoli (nucleolus is singular). The individual chromosomes are not visible during this stage but are replicated in S phase. This replication produces a pair of identical chromatids held together by a centromere. Figure 2A Identify a cell in the onion root tip that is in interphase Prophase (Figure 2B). The chromosomes, now containing two chromatids joined by a centromere, condense and become visible as dark linear structures. The spindle begins to form. You should be able to see the chromosomes but neither the chromatids nor the centromere will be visible at this magnification. Figure 2B Identify a cell in the onion root tip that is in prophase 2
3 Prometaphase (Figure 2C). During this stage, the nuclear envelope will be disassembled and the nucleolus will no longer be visible. The spindle continues to form and the spindle fibers begin to attach to the chromosomes. Figure 2C Identify a cell in the onion root tip that is in prometaphase and Metaphase (Figure 2D). The spindle apparatus is completely formed and spindle fibers have attached to each chromosome at the centromere. This causes the chromosomes to become aligned on the equator. You should be able to see the spindle fibers. Make sure you identify the chromosomes and the equator. Figure 2D Identify a cell in the onion root tip that is in metaphase Anaphase (Figure 2E). The centromere divides allowing the sister chromatids to begin to move apart. At this point since the chromatids have separated and each has its own centromere they are each called a chromosome. The chromosomes have begun to move to their respective poles. Be sure to identify the chromosomes and the spindle. Figure 2 E Identify a cell in the onion root tip that is in anaphase 3
4 Telophase (Figure 2F). Once the chromosomes have reached their respective poles they will uncoil and gradually become less visible. The nuclear envelope will be reformed around each complete set of uncoiling chromosomes. The nucleolus will once again become visible. Look for a cell in which the chromosomes have arrived at their respective poles. Cytokinesis (Figure 2F). During telophase a cell plate will form from the center of the equator outward separating the cytoplasm of the two new daughter cells. When the cell plate has completely divided the two daughter cells they are considered to be in interphase. Look for cells in late telophase where the cell plate should be visible and the chromosomes are being reorganized into a nucleus. Figure 2F Identify a cell in the onion root tip that is in telophase/cytokinesis and B) Identify all the different stages of mitosis in the diagram below. Circle and label Figure 3 below Fig 3: Mitotic stages in Onion root tip cells (under microscope, 400X view) 4
5 Activity 2 Mitosis in Animals A) The same mitotic stages exist in animals as in plants. To observe stages of animal mitosis use the whitefish blastula slide under low power and locate a group of dividing cells. Focus it on high power and notice that the cells are less ordered than you saw in the onion root tip. This may allow you to view mitosis from several angles causing some stages to look atypical. Look for cells that are more typical in appearance. You have several groups of cells to look at so if you can t find a particular stage in the group you are in use the mechanical stage to move to another group of cells. Draw each phase in the space provided labeling any structures indicated in bold print. Figures 4A-2F are model representations of various stages. Interphase. This stage will be similar to what you saw in the plant cell but you may not be able to see the nucleolus and there is no cell wall present. blastula that is in interphase Prophase. The chromosomes become visible as thread like structures. You will notice that the chromosomes are smaller and more numerous than in the plant cell. Two cytoplasmic structures not found in the plant cell called centrioles have migrated to opposite poles and are beginning to align the microtubules of the spindle. While each centriole is not visible the microtubules migrating out from it will be visible and have a star like appearance. These star like structures are asters (a cluster of microtubules that radiate out toward the poles when a dividing cell has centrioles). Make sure you can also locate the chromosomes and the spindle fibers. blastula that is in prophase and 5
6 Prometaphase (Figure 4A) The nuclear membrane and the nucleoli disassemble. The asters move towards opposite poles of the cell as the spindle fibers begin attaching to chromosomes. Figure 4A blastula that is in prometaphase and draw what you see here. Metaphase (Figure 4B) Events in this stage are very similar to the onion root tip. One difference you might note is that the spindle and its individual fibers are easier to see. Be sure to identify the aster, spindle fibers, chromosomes and equator. Figure 4B blastula that is in metaphase Anaphase (Figure 4C and 4D) Events here are like in the onion root tip. Identify the aster, spindle, spindle fibers, and chromosomes. Figure 4C blastula that is in anaphase and Figure 4D 6
7 Telophase and cytokinesis (Figure 4E and 4F) Unlike the onion root tip no cell plate forms during cytokinesis. Instead the plasma membrane appears to pinch in on either side of the equator forming a V like structure called the cleavage furrow. This will continue to deepen until it divides the cell into half to form two identical daughter cells. Each daughter cell is about half the size of the parent cell. blastula that is in telophase/cytokinesis and Fig 5: Mitotic stages in whitefish blastula (under microscope, 400X view) (Picture courtesy: "BaileyBio.com." RSS. N.p., n.d. Web. 26 Apr. 2014) 7
8 Pre-Lab Activity Prior to this lab you should: 1. Read section 12.2 from your textbook (Freeman, 6th edition; pages ) 2. Define the following phases and structures in the table below Definition Cell Cycle G1 Phase G2 Phase S Phase M phase Chromosome Chromatid Centromere Centrosome Cytokinesis Cell Plate Cleavage Furrow Spindle 8
9 Post Lab Activity Complete the following questions: 1. What differences are there between plant cell and animal cell division? 2. Describe each stage of cell division in plant cells. 3. Describe each stage of cell division in animal cells. 4. In which stage of the cell division in animal cells will asters appear? 5. In which stage do the chromosomes line up at the equator? 6. In which stage will sister chromatids separate? 7. In which stage does the nuclear envelope reform? 8. What structure holds the individual chromatids together? 9. How does cytokinesis occur in plant cells? How does it occur in animal cells? 10. What occurs during each part of interphase? 11. What is the purpose of cell division? 12. Each chromosome is formed by two 13. After mitosis is completed, two daughter cells are produced from a diploid mother cell 14. Identify the stages in the following diagram and arrange them in order 15. Above stages of cell division are in a. animal cells b. plant cells 9
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