Contributions of Fruit Flies in Life Science Research

1. 模式动物果蝇对生命科学研究的重要贡献 概述: 1，果蝇在生命科学研究中的历史贡献 2，来源于果蝇研究中的重大技术突破 3，果蝇在现代医学研究中的应用 专题：突触发育与智障和自闭症

2. 成虫 4d 胚胎 1d 蛹 10hrs 1.5d 1d 2d 幼虫 ~10 days at 250C

3. Most Commonly Used Mutants wildtype, WT white yellow Curly Bar ebony

4. Basic Genetics Concepts Mutants--lots and lots of various mutants Mutations--loss-of-function and gain-of-function mutations Mutagen--chemical, physical and biological mutagens Mutagenesis--generation of various mutants Mosaics--mutant tissues or cells in otherwise wildtype individual Epistasis--gene interaction in terms of phenotypes Genotype versus phenotype Forward genetics versus Reverse Genetics

5. Genetics Versus Biochemistry Biochemistry: the scientific study of the chemistry of living cells, tissues, organs and organisms; in vitro structural and functional study of purified living components Genetics: the study of the patterns of inheritance of specific traits; understanding the in vivo functions of a gene through mutants Interdisciplinary approach to study biological processes Protein: 蛋白、蛋白质 Mutation/mutants: 突变、突变体

6. The Nobel Prize in Physiology or Medicine 1933 For his discoveries concerning the role played by the chromosome in heredity Morgan, T.H. Science 32: 120-122 (1910) Sex limited inheritance in Drosophila From Columbia University to California Institute of Technology

7. Morgan and His Three Extremely Gifted Students Alfred H. Sturtevant Calvin B. Bridges Hermann J. Muller

8. A. H. Sturtevant

9. The Nobel prize in Physiology or Medicine, 1946 For the discover of the production of mutations by means of X-ray irradiation Hermann Joseph Muller, 1890-1967 Indiana University, Bloomington, IN, USA Before his discovery, mutants were largely from spontaneous mutations!! It revolutionized animal and plant breeding, offered novel insights into irradiation damage and cancer biology

10. Ed B. Lewis, 1918-2004 Nobel Prize Winner in Physiology or Medicine, 1995 Lewis, E.B. (1978) A Gene Complex Controlling Segmentation in Drosophila. Nature 276, 565-570

12. Two-headed lamb born at a family farm in New Zealand Oct. 10, 2014

13. Homeotic gene expression--colinearity All transcription factors

14. The Nobel Prize in Physiology or Medicine, 1995 For their discoveries concerning the genetic control of early embryonic development Edward B. Lewis Christiane Nusslein-Volhard Eric F. Wieschaus 77 52 48 Nuesslein-Volhard, C., Wieschaus, E. (1980). Mutations Affecting Segment Number and Polarity in Drosophila. Nature 287, 795-801 Some of these genes may be responsible for spontaneous abortions and congenital malformations

15. Walter Gehring, University of Basel, Switzerland Those who succeed… …are the most hard-working and persistent. EMBL, Heidelberg

16. A Turning Point in the Historyof Developmental Genetics The October 30, 1980 cover “Mutations Affecting Segment Number and Polarity in Drosophila”

17. Classical F3 Screen for Recessive Mutations EMS propagation Self-cross Balanced homozygotes: Dia as larvae Balanced mutant stock: viable Homozygous for mutations: Screen embryos for phenotypes Tedious and time-consuming: single crosses, three generations

18. Mutations and Genes Identified from the Screen WT wingless staufen

19. 150 development-regulating genes that affect gross morphology in Drosophila stimulated the search for development genes in other systems (nematode and mouse) virtually all the genes involved in development of Drosophila are represented also in vertebrates, conserved over 700 million years of evolution

20. The Uniqueness of the Nobel Prize Screen 1, Screen for phenotypes in embryo instead of adult (1965-1980) 2, the first screen to find mutations affecting a given process Developmental features crucial for the success of the screen Larval cuticle: an excellent readout of embryo patterning Embryogenesis is very fast, housekeeping proteins are provided from the egg. Only the patterning genes are specifically identified.

21. The Nobel Prize in Physiology or Medicine, 1995 For their discoveries concerning the genetic control of early embryonic development Edward B. Lewis Christiane Nusslein-Volhard Eric F. Wieschaus Determined to understand a biological process and be persistent! We were young and foolish, and it was worth trying

22. The Noble Assembly at Karolinska Institut has today decided that the Noble Prize in physiology or medicine shall be divided, with one half jointly to Bruce Beutler and Jules Hoffmann for their discoveries concerning the activation of innate immunity and the other half to Ralph M. Steinman for his discoveries of the dendritic cell and its role in adaptive immunity Jules Hoffmann 2011-10-03

24. Conserved pathways in innate immunity Hoffmann et al., Science, 1999

25. Illustration by Mattias Karlen

26. 2011 Noble Prize work on immunology

27. Thomas Hunt Morgan Medal Genetics Society of America Medal George W. Beadle Medal 1983 1995 7 Nobel Prize Winners 1958 2003 1995 2003 2003

28. Thomas Hunt Morgan Medal Genetics Society of America Medal George W. Beadle Medal 12 recipients

29. Michael Ashburner Gene regulation: chromosomal puffs Books: most popular books Genome: successful fly genome projects Database: most comprehensive flybase Department of Genetics Cambridge University

30. David Hogness: The 2003 Thomas Hunt Morgan Medal Department of Biochemistry, Stanford University David Hogness and son, Peter

31. David Hogness: The 2003 Thomas Hunt Morgan Medal application of mol. biol. to fly genetics 1, produced first genomic library of any organism 2, Established the method of “positional cloning” or “map-based cloning” 3, Establsihed “colony hybridization”, 1975 4, First person to practice “genomics” and “functional genomics” 5, Discovered “Goldberg-Hogness box”, now known as the TATA box, 6, Trained many well-known scientists, eg., Gerry Rubin, Richard Mann

32. Two Most Active Players in the Fly Community in the Last Twenty Years Gerry M. Rubin UC Berkeley Vice President of the HHMI Allan C. Spradling, Director of the Department of Embryology of the Carnegie Institute Rubin, G.M., Spradling, A.C. Genetic transformation of Drosophila with transposable element vectors. Science, 1982 218(4570):348—353 Prominent leaders in the Drosophila Genome Project Gerry: the Berkeley Drosophila Genome Project, sequencing of genomic clones, EST clones and mapping of P element insertions. The genome sequence of Drosophila melanogaster. Science2000 287(5461):2185--2195 Allan: Genetic screens to mutate the genome by P elements, aiming at having a P insertion in each of the 15,000 genes. The BDGP gene disruption project: single transposon insertions associated with 40% of Drosophila genes.

33. 模式动物果蝇在生命科学和医学研究中的应用 1，果蝇在生命科学研究中的历史贡献 2，来源于果蝇研究中的重大技术突破 (P Insertion; UAS-GAL4; FRT-FLP) 3，果蝇在现代医学研究中的应用

34. Landmarks in Fly Genetics • Elucidation of early development by systematic genetic screens, 1980; Nobel Prize in 1995 • Germline transformation of fruit flies, 1983 • Completion of Genome Sequencing in 2000, fly as a model for human diseases • Mutagenesis by P element insertion, each gene has a insertion • UAS-GAL4 system to manipulate gene expression • FRT-FLP system to make mosaic clones

38. Gene Expression Manipulation UAS-GAL4 System Derived From Yeast Brand, A.H., Perrimon, N. Targeted gene expression as a means of altering cell fates and generating dominant phenotypes. DEVELOPMENT 1993 118(2):401--415 Cited: 5983 Rorth, P. A modular misexpression screen in Drosophila detecting tissue-specific phenotypes. PNAS 1996 93(22):12418--12422 Rorth, P., Szabo, K., Bailey, A., Laverty, T., Rehm, J., Rubin, G., Weigmann, K., Milan, M., Benes, V., Ansorge, W., Cohen, S. Systematic gain-of-function genetics in Drosophila. DEVELOPMENT 1998 125(6):1049--1057

39. GAL4–UAS System for Directed Gene Expression GAL4: a transcription factor UAS: upstream activating sequence Yeast: Nature, 1988 Cre floxP

40. Cre-LoxP system in mammals From www.wikipaedia.org

41. Chromosomal Recombination—Mosaic Analyses FRT: a specific DNA sequence FRT-FLP System FLP: flippase or FLP recombinase Xu, T., Rubin, G.M. Analysis of genetic mosaics in developing and adult Drosophila tissues. DEVELOPMENT 1993 117(4):1223--1237 Cited: 1600 Lee, T., Luo, L.Mosaic analysis with a repressible neurotechnique cell marker for studies of gene function in neuronal morphogenesis. NEURON 1999 22(3):451--461 Cited: 1279 Mouse: Cre-LoxP System from bacteriophage P1

42. Mosaic Analyses—Powerful Genetic Toolconditional knockout Daughter cells DNA duplication No recombination 偷梁换柱 Recombination induced by yeast derived Flipase and FRT sequences

43. Advanced Mosaic Analysis--MARCM Lee T & Luo L, Neuron, 1999 Mutant, no label classical WT, labeled Mutant, labeled MARCM WT, no label

44. Single Brain Neuron When FMRP Expression is Altered WT Mutant Overexpression

46. Bellen et al., Nat Rev Neurosci, 2010

47. Bellen et al., Nat Rev Neurosci, 2010

48. 模式动物果蝇在生命科学和医学研究中的应用 1，果蝇在生命科学研究中的历史贡献 2，果蝇研究中的重大技术突破 3，果蝇在现代医学研究中的应用

49. 果蝇在医学研究中的应用 1, Infection and immunology 2, Cancer biology and signal transduction 3, Neurobiology: ion channels, circadian rhythm, learning and memory 4, Neurological diseases: neurodegenerative disease and mental retardation

50. A Good Model to Study Cancer Biology