1. Using a diagram, explain how MCSF facilitates the development/differentiation of macrophages from progenitor cells.
2. List/explain 2 key differences between the fish and the mammalian MCSFs.
3A) Diagram how the single mammalian MCSF gene produces multiple MCSF isoforms.
3B) Draw and describe 3 of these isoforms in terms of their protein structure and post-translational modifications.
4. Draw a phylogenetic tree of vertebrate MCSF proteins. Include in this tree only the following:
mouse MCSF, human MCSF, zebrafish MCSF1, zebrafish MCSF2, salmon MCSF1, salmon MCSF2 and predict where the frog and the chicken MCSFs would branch.
5A. Are the different fish MCSF gene sequences more closely related to each other or to the mammalian MCSF gene sequences?
5B. According to the authors, what does this suggest about the functional roles of the fish MCSF molecule?
5C. Explain why you agree or disagree with the authors in this regard.
6. How (what technique) would you use to determine whether the trout MCSF1 undergoes alternative splicing? Briefly explain how this would indicate the presence or absence of splice variants.
7. Explain what is the technique used in Figure 7, how does it work and whether and why it is more or less sensitive than the technique.
8. What is the technique used in Figure 10B? Using what you learned in class, explain how this technique works.
9. Use your reasoning and description of the technique used in Fig. 10B to explain the meaning behind the difference in the responses elicited by MCSF1/purified versus CCM.
10. What are RTS-11 cells and why do you think they were used for the studies in this paper?
11. List four differences in the gene expression of the trout MCSF1 and MCSF2.
12A. Explain what is PMA and why was it used in this paper in terms of what you learned in class.
12B. Briefly explain whether the results depicted in Fig. 9A are consistent with what the authors expected to see based on what is known to happen in mammals.
12C. Based on your answer in 12A, how could the authors confirm that PMA is acting on the fish cells the way that they expect it to?
13. What is the role of the fish head kidney and what tissue/organ performs this role in human?