BIOL 212 Genetics Lab Spring 2007 Lab 4: Mitosis and Meiosis Purpose: To review the stages of mitosis (somatic cell division) and meiosis (cell division in gametes). An understanding of these basic processes is fundamental to the study of Mendelian genetics. Materials: Laptop computers/internet access Slides, compound microscopes, lens paper Mitosis slides: whitefish blastula (Leucichthys blastodisc) Meiosis slides: Ascaris or Parascaris (nematode) oogenesis (slides #M1194, M1195, M1196) Colored pencils Index cards Background: Hartl and Jones, Essential Genetics, 4th ed. Chap. 3 pp. 76-88. Procedure: 1. A. Practice working through the processes of mitosis and meiosis for a diploid cell containing 2 or 3 pairs of homologous chromosomes. You can do this by color-coding the drawings of meiosis and mitosis on the last pages of the handout using color pencils, or by working through examples from the following web site: http://biog-101-104.bio.cornell.edu/BioG101_104/tutorials/cell_division/CDCK/cdck.html In genetics, we will focus on what the chromosomes are doing during cell division, rather than on the changes in cell structure that are also taking place. Ask yourself the following questions as you work through mitosis and meiosis. What is the number of chromosome sets at each phase (haploid=1N or diploid=2N)? When does DNA replication take place? How much DNA is present at each stage in comparison to a nondividing cell of the same type? I prefer to use the symbol “C” to refer to DNA content separately from chromosome content (i. e. haploid or diploid). Thus a nondividing diploid cell would have 2C DNA, a diploid cell which has undergone DNA replication prior to cell division has 4C DNA (yet is still 2N). Gametes are haploid (1N) and contain 1C DNA. B. You will be assigned one of the following stages of mitosis or meiosis: 1 Interphase, Prophase, Metaphase, Anaphase, or Telophase of mitosis Interphase, Prophase I, Metaphase I, Anaphase I, Telophase I, Prophase II, Metaphase II, Anaphase II, Telophase II of meiosis. Prepare to present in class a cell with a given chromosome set that is in that stage of cell division. 2. Review the process of mitosis using the diagrams in Fig. 2 of the handout. Observe mitosis on prepared slides of whitefish blastula under the light microscope. Identify cells in the stages of prophase, metaphase, anaphase, telophase and interphase. See instructor for help with setting up microscopes or locating structures. 3. Review the process of meiosis in Ascaris using diagrams B through E in Fig. 4. In Ascaris, penetration by the sperm initiates meiosis in the oocyte. Note that in oogenesis, only one of four gametes produced in meiosis survives to be fertilized. Other genetic material is expelled as polar bodies. Once meiosis is complete in the oocyte, the gametes (male and female pronuclei) fuse. Observe oogenesis in Ascaris. Examine a slide containing sections of the Ascaris uterus with oocytes in various stages of maturation. Ascaris contains four large chromosomes (2 homologous pairs). Some terms used in diagrams/descriptions of Ascaris meiosis: tetrads: 4 duplicated chromosomes (the two homologous pairs after replication) dyads: 2 duplicated chromosomes (one from each homologous pair) polar body: genetic material that is discarded after each meiotic division in the oocyte pronucleus: male and female haploid nuclei prior to fertilization Students will be assigned to identify a particular stage of Ascaris meiosis and to set up one of the microscope stations for the class. Using Fig. 4 as a reference, try to identify the following stages on the prepared slides set up on the demonstration microscopes (B-E refer to the labels in Figure 4). Please consult the instructor for help in identifying stages. Station #1: Station #2 Station #3 Station #4 Station #5 Station #6 Station #7 unfertilized primary oocytes (B), sperm entrance (B), oocyte with tetrads in meiosis I (C), formation of first polar body (C), secondary oocyte with dyads in meiosis II (D), formation of second polar body (D&E), male and female pronuclei (E). When all the stations are set up, try to visit them in sequence to better understand the process of meiosis in Ascaris. 2 3 4 5 6 Quiz: There will be a quiz next week in lab (20 pts.) following a short discussion. The quiz will emphasize what happens to the chromosomes during mitosis and meiosis and some of the major distinctions between the two processes. It would be good practice to use the “random” feature of the Cell Division Construction Kit http://biog-101-104.bio.cornell.edu/BioG101_104/tutorials/cell_division/CDCK/cdck.html for practice in visualizing the number and arrangement of chromosomes in cells in different stages of cell division. Some questions may also be drawn from the slides which you observed. A sample quiz will be posted on-line. 7