Biol 441 Review questions for techniques:

1. What did the Acetabularia experiments tell us about the importance of nucleus or cytoplasm in differentiation?

2. What conclusions can arise from the work on amphibian nuclear transplantation? What is different about the work of Briggs and King vs. Gurdon?

3. What can you say about the effects of egg cytoplasm on different nuclei transplanted in? What new activities can those nuclei do they didn't do in the cells they came from?

4. What general kinds of nucleic acid hybridization experiments can be done (isolate mRNA, make cDNA and clone to use as reagent, hybridize with whole genome DNA or RNA or tissue in situ) or and what are the requirements (Labeling with nucleotides, melt, renaturation, electrophoresis, transfer by blot, enzyme breakdown of non-paired. Learn an example of DNA-DNA and one of DNA-RNA hybridization.

5. Learn Ways to detect gene activity:

Histochem- look at chromatin for ruffing, look for RNA production using radioactive nucleotides;DNA RNA in situ hybridization

6. Learn ways to compare RNA between stages, using hybridization

7. look for protein production-electrophoresis

8. How to detect gene control proteins:

Footprinting

Inject into egg with gene

9. How can making a reporter gene construct with a different control region be useful in study of gene control?

10. How are restriction enzymes useful in making such constructs?

11. What would a sea urchin blastula cDNA library be?

12. What can you say about the dot blot on pg 65? For examp[[le, HOw is clone D42 different from DG 72 or r5?

13. How would you describe the difference between the adhesive behavior of ectoder, mesoderm, and endoderm?

14. How do different cell types become segregated from each other? What are the membrane structural molecules which interact to cause cell adhesion?

15. Distinguish between CAMs and Cadherens.

17. What is the difference between cell-cell adhesion and cell -ECM adhesion?

18 What kinds of reactions due to signal transduction occur in both sperm and eggs in spite of different sets of receptors in each?

19. How does the amount of yolk influence development?

20. What is different between amniotes and anamniotes?

21. What is different about signals for meiosis to occur and mitosis after fertilization?

22. How can antibodies be used to detect different receptors or surface molecules on different parts of cells or different kinds of cells?

23. What is the difference between the fast block to polyspermy and the slow block?

24. How are the acrosomal reaction and the cortical reaction similar? Both can only happen once and then the gametes will die if they are not together as a result of those reactions: think about what reacted sperm can do if placed with new eggs, and what can reacted eggs do if placed with new sperm.

25. What is the significance of cytoplasmic movement or shifts after fertilization? Give some examples and how you can prove something significant happened.

26. How can fate maps be drawn and what is the relationship between fate maps of different stages?

27. How are movements of the three germ layers different from each other?

28. How is formation of inner cell mass and then cavity formation in mammals connected with extraembryonic membranes and the primitive streak of the embryo?

29 How are the cell movements or other mechanisms of neurulation different from gastrulation?

30. What are the brain regions and how are they related to notochord and somites and prechordal plate? What is the difference between structures derived from basal and alar plates and neural crest? What are some differences in Pax genes found in those regions?

31. How is a reflex arc constructed in the embryo?