1. Lecture 1 Course Outline Central problem in Development Terms Cellular mechanisms of determination

2. Objectives Lectures: application of genetic analysis to problems in development. Emphasis on problem solving as opposed to rote memorization. Labs: Illustration of some basic techniques used in Developmental genetics. Lab report written in the format of a scientific paper.

3. Jan 12 Lecture 1 Introduction to developmental genetics Jan 19 Lecture 2 Introduction to genetic dissection: screens, complementation and epistasis. Jan 26 Lecture 3 Drosophila cell biology Feb 2 Lecture 4 Modern Drosophila genetics Feb 9 Lecture 5 Anterior posterior axis formation Feb 16 Lecture 6 Anterior posterior axis formation continued Feb 23 SLACK WEEK March 1 Lecture 7 Morphogens March 8 Lecture 8 Nuclear gradients and the habits of developmental signaling pathways March 15 Lecture 9 Short course on C. elegans cell biology and genetic analysis. March 22 Lecture 10 Lateral inhibition—emergent properties in development March 29 Lecture 11 Short course on Evolution and Development. April 5 Lecture 12 Experimental approaches to Evo Devo. Paper due. Schedule of Lectures

4. Date for the Midterm examination Monday February 27, 2012 This is the Monday right after slack week 7-10PM. This midterm will cover material up to Lecture 6 Anterior Posterior axis.

5. The course web site can be found at instruct.uwo.ca/biology/4540g This web site contains all the information you need for this course. Consult it. Explore it.

6. Jan 9/10 NO LAB Jan 16/17 Lab 1 Preparation of first instar larval cuticles (test) Jan 23/24 Lab 2 Ectopic expression of Fushi tarazu (come in the day before). Fixation of embryos for Labs 3&4 (test) Jan 30/31 Lab 3 Engrailed antibody staining (long lab) (test) Feb 6/7 Lab 4 Wingless in situ hybridization (come in the day before; long lab) (test) Feb 13/14 Lab 5 Beta-galactosidase staining for assaying ftz enhancer activity. (test) NO MORE LABS Schedule of the Labs and in class tests

7. Jan 9/10 NO LAB Jan 16/17 Lab 1 Preparation of first instar larval cuticles Jan 23/24 Lab 2 Ectopic expression of Fushi tarazu (come in the day before). Fixation of embryos for Labs 3&4 Jan 30/31 Lab 3 Engrailed antibody staining (long lab) Feb 6/7 Lab 4 Wingless in situ hybridization (come in the day before; long lab) Feb 13/14 Lab 5 Beta-galactosidase staining for assaying ftz enhancer activity. NO MORE LABS Long labs

11. The lab test on the readings Why am I doing this to you? In the past I expected students to read the papers on their own before coming to the lab. Over the years it become clear that this was not occurring. Reading in the first half of the term saves a lot of pain later. Since I know this, I am being a pain now.

12. Format of the test on the readings You will have 5 minutes to answer one question at the beginning of the lab. The question asked will be different on Monday and Tuesday.

13. This and the next lecture will focus on definition of the terms and basic genetic concepts used in Developmental genetics.

14. The central problem in Development “How is the information stored in linear DNA transformed over time into a three dimensional multicellular organism?”

18. From this perspective the problem seems limitless. However, the size of the genome provides a limit on the problem because all information required for hand development is somehow encoded in the genome.

21. What is animal development?

22. It is an essential part of our and all animal life cycles.

23. What is animal development? It is an essential part of our and all animal life cycles. At the present time only life can beget life.

24. General animal life cycle Soma Germ-line fertilization

25. General animal life cycle Soma Germ-line fertilization Somatic germ-line division

26. General animal life cycle Soma Germ-line fertilization Somatic germ-line division Life does not begin at fertilization; it is a continuous process.

27. What are the characteristics of animal development?

28. Development at its most basic level. Single celled Zygote Decisions Complex multicellular organism of many specialized cell types and organs.

29. Two terms used in Developmental Biology. Determination Differentiation

30. More terms Cell specification versus Pattern formation

31. Examples of cell specification Lens: each cell as it differentiates becomes transparent. Red blood cell: no nucleus packed with hemoglobin.

33. Example of pattern formation You distinguish an arm from a leg by its shape and not by what it is made up of which is basically the same stuff: bone, muscle, nerves etc.. This is an abstract ability, because shape is a quality defined only by what it is.

34. Cellular mechanisms of determination Intrinsic Extrinsic

35. mRNA protein Anterior Posterior Example of an intrinsic mechanism: Assymetric determinant Bicoid mRNA and protein are assymetrically localized in the cell.

36. Extrinsic mechanisms A) Lateral inhibition B) Induction

37. Lateral inhibition during neurogenesis in Drosophila. Example of an emergent property because before the process starts you can not tell which cell will be neurogenic and which will be epithelial.

38. Delta ligand Notch receptor Start off with identical levels of expression in two cells.

39. Delta ligand Notch receptor Start off with identical levels of expression in two cells. No Net signal

40. Due to a stochastic (random, chance) event one cell expresses more Delta ligand than the other. The net signal is in this direction. The activation of Notch receptor results in decreased Delta expression. Dl

41. When Notch or Delta are absent, the ectoderm cells become neurogenic. The cell that becomes the neuroblast inhibits the surrounding cells from this fate, and telling them to become epithelial cells instead. Lateral inhibition. NeuroblastEpithelial Notch Neuroblast Epithelial

42. Induction 1 2 A cell or group of cells removed from a second cell that directs the developmental fate of a second cell or group of cells.

43. Example of induction Anchor cell-gonad signals Epidermis Vulva

44. What molecules control determination?

46. Gut Brain Analysis of gene regulation: cis regulation. gene something embryo of something ORF

47. ATG Translational fusion to a reporter gene Reporter gene

48. ATG GFP TATA Regulatory regions of the gene

49. ATG GFP TATA Regulatory regions of the gene Gut Brain

50. ATG GFP TATA Regulatory regions of the gene Gut Brain Gut Brain

51. ATG GFP TATA ATG GFP TATA ATG GFP TATA ATG GFP TATA ATG GFP TATA Mutagenesis Gut Brain Gut Brain

52. ATG GFP TATA ATG GFP TATA ATG GFP TATA ATG GFP TATA ATG GFP TATA Mutagenesis Gut Brain Gut Brain Gut Brain

53. ATG GFP TATA ATG GFP TATA ATG GFP TATA ATG GFP TATA ATG GFP TATA Mutagenesis Gut Brain Gut Brain Gut Brain

54. ATG GFP TATA ATG GFP TATA ATG GFP TATA ATG GFP TATA ATG GFP TATA Mutagenesis Gut Brain Gut Brain Gut Brain Gut Brain

55. ATG GFP TATA ATG GFP TATA ATG GFP TATA ATG GFP TATA ATG GFP TATA Mutagenesis Gut Brain Gut Brain Gut Brain Gut Brain

56. ATG GFP TATA ATG GFP TATA ATG GFP TATA ATG GFP TATA ATG GFP TATA Mutagenesis Gut Brain Gut Brain Gut Brain Gut Brain Gut Brain

57. ATG GFP TATA ATG GFP TATA ATG GFP TATA ATG GFP TATA ATG GFP TATA Mutagenesis Gut Brain Gut Brain Gut Brain Gut Brain Gut Brain

58. ATG GFP TATA ATG GFP TATA ATG GFP TATA ATG GFP TATA ATG GFP TATA Mutagenesis Gut Brain Gut Brain Gut Brain Gut Brain Gut Brain Gut enhancer Brain enhancer Gut silencer

59. ATG Heterologous TATA box Transcriptional fusion Gut Brain Gut enhancer Brain enhancer Gut silencer GFP

60. ATG GFP Gut Brain

61. ATG GFP Gut Brain ATG Gut enhancer Brain enhancer Gut Brain Gut Brain

62. ATG GFP Gut Brain ATG Gut enhancer Brain enhancer Gut Brain Gut Brain

63. ATG GFP Gut Brain ATG Gut enhancer Brain enhancer Gut Brain Gut Brain ATG Gut enhancer Gut silencer Gut Brain

64. ATG GFP Gut Brain ATG Gut enhancer Brain enhancer Gut Brain Gut Brain ATG Gut enhancer Gut silencer Gut Brain

65. Important property: Necessary and Sufficient

66. Developmental pathways

67. Transcription factor signal transcription factor pathways