1. Lecture 5 Anterior Posterior axis formation: Cell Biology Bicoid is a morphogen

2. Lab 5 schedule On Monday and Tuesday come in at the regular time. On Tuesday and Wednesday come to B&G 3015 to mount your embryos. Elyse will discuss the time of day to come.

3. Exams Midterm Tuesday Feb. 28, 2012 UCC 63, 7-10PM Tentative Final exam date Thursday April 19, 2012 at 9:00AM

4. Structure of the egg Dorsal appendage Chorion Vitelline Micropyle D V A P

5. Cellular Blastoderm Hartenstein 1993

6. Blastoderm fate map Hartenstein 1993

7. Simplified Fate Map Nusslein-Volhard et al., Science 238, 1675-1681

8. Remove posterior pole plasm Classical Embryology Remove 5- 10% of the anterior cytoplasm Nusslein-Volhard et al., Science 238, 1675-1681

9. Transplant anterior cytoplasm to posterior Nusslein-Volhard et al., Science 238, 1675-1681

10. Transplant posterior cytoplasm to anterior Nusslein-Volhard et al., Science 238, 1675-1681

11. Transplant posterior cytoplasm to anterior Nusslein-Volhard et al., Science 238, 1675-1681

12. Transplant posterior cytoplasm to an anterior that lacks the anterior signal Nusslein-Volhard et al., Science 238, 1675-1681

13. Properties of the polar center signals Both exert long range effects The anterior and posterior activities inhibit each other Both activities have inductive properties

14. Properties of the polar center signals Both exert long range effects

15. Remove posterior pole plasm Classical Embryology Remove 5- 10% of the anterior cytoplasm Nusslein-Volhard et al., Science 238, 1675-1681

16. Properties of the polar center signals The anterior and posterior activities inhibit each other

17. Transplant posterior cytoplasm to anterior Nusslein-Volhard et al., Science 238, 1675-1681

18. Transplant anterior cytoplasm to posterior Nusslein-Volhard et al., Science 238, 1675-1681

19. Properties of the polar center signals Both activities have inductive properties

20. Transplant anterior cytoplasm to posterior Nusslein-Volhard et al., Science 238, 1675-1681

21. Transplant posterior cytoplasm to an anterior that lacks the anterior signal Nusslein-Volhard et al., Science 238, 1675-1681

22. What you may know already Bicoid: Anterior signal Nanos: Posterior signal How do we know this?

23. Maternal effect screens bcd/bcd mother Normal egg Larva missing a head

24. Nusslein-Volhard et al., Science 238, 1675-1681

25. Three classes of maternal effect phenotypes Nusslein-Volhard et al., Science 238, 1675-1681

26. The real cuticles wt bcd oskar torso Nusslein-Volhard et al., Science 238, 1675-1681

27. Mutant combinations St Johnston and Nusslein-Volhard Cell 1992

28. Transplantation of cytoplasm into bcd mutants Nusslein-Volhard et al., Science 238, 1675-1681

29. Controlling anterior posterior polarity Nusslein-Volhard et al., Science 238, 1675-1681

30. The difference between the posterior and anterior signals Nusslein-Volhard et al., Science 238, 1675-1681

31. Distinct Properties of the signals Anterior signal-instructive. Induces polarity at site of injection. Posterior signal-permissive. Polarity independent of injection site.

32. Evidence for Bicoid Rescue activity spatially distributed Amount of rescue dependent on amount injected Linear relationship between gene dose and rescue activity mRNA asymmetrically localized and protein expressed in a gradient

33. Bicoid rescue assay Nusslein-Volhard et al., Science 238, 1675-1681

34. % rescue activity 100% 50% % egg length 100% 0%40% Rescue activity is spatially distributed

35. % rescue activity 01/32/31 Amount of rescue is dependent on amount injected 100% 50% Transplanted volume

36. Linear relationship between gene dose and rescue activity 100% 50% 012 3 % rescue activity Dose of bicoid gene in mothers

38. mRNA assymetrically localized and protein expressed in a gradient mRNA Protein Driever and Nusslein-Volhard Cell 54, 83-93

39. Bicoid Bicoid has the properties of the anterior signal Does Bicoid need to interact with any other anteriorly localized signals to work?

41. Bicoid is necessary and sufficient Bicoid mRNA made in vitro mRNA injected at posterior pole Bicoid protein expression Larva with two heads Driever et al., 1990 Development 109, 811-820

42. Bicoid Bicoid is the anterior signal What does Bicoid encode? How does Bicoid work?

43. Bicoid encodes a homeodomain containing protein.

44. Hunchback lacks anterior segments Nusslein-Volhard and Wieschaus 1980 Nature 287, 795

45. Bicoid mRNA Bicoid protein Hunchback mRNA expression Hunchback is expressed at the anterior pole Driever 1993

46. Hunchback gene Driever and Nusslein-Volhard Nature 337, 138-143

47. Bicoid is a transcriptional activator Driever and Nusslein-Volhard Nature 337, 138-143

48. mRNA assymetrically localized and protein expressed in a gradient mRNA Protein Driever and Nusslein-Volhard Cell 54, 83-93

49. Morphogen Long range action Affect polarity Concentration dependent Different fates at different concentrations

50. Morphogen Long range action

51. Bicoid mRNA Bicoid protein Hunchback mRNA expression Hunchback is expressed at the anterior pole Driever 1993

52. Morphogen Affect polarity

53. Bicoid is necessary and sufficient Bicoid mRNA made in vitro mRNA injected at posterior pole Bicoid protein expression Larva with two heads Driever et al., 1990 Development 109, 811-820

54. Morphogen Concentration dependent

55. Concentration dependence of Bicoid function

56. # of bicoid genes in the mother 1 2 0 4 6 hb-lacZ expression Concentration dependence of Bicoid function Struhl et al., 1989 Cell 57, 1259-1273

57. Morphogen Different fates at different concentrations

58. Distinct Bicoid binding sites are activated at different concentrations of Bicoid

60. Morphogen Long range action Affect polarity Concentration dependent Different fates at different concentrations

61. How is bicoid mRNA localized to the anterior pole?

63. Other anterior class mutants Driever and Nusslein-Volhard Cell 54, 94-104

64. Bicoid gradient affected in the mutants Driever and Nusslein-Volhard Cell 54, 94-104

65. The mutants affect different stages of bicoid mRNA deposition in the developing oocyte St. Johnston et al., 1989 Development sup 13-19

66. Localization of bicoid mRNA: recent paper Irion and St Johnston 2007 Nature 445, 554-558