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    Why Use Drosophila To Study Human Disease?

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    Daiana Groisman

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    Why Use Drosophila To Study Human Disease?

    Uploaded by

    Daiana Groisman
    25 views41 pages

    Original Title

    2IntroToDrosophila

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    © © All Rights Reserved

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    25 views41 pages

    Why Use Drosophila To Study Human Disease?

    Uploaded by

    Daiana Groisman

    Copyright:

    © All Rights Reserved
    Download as PPTX, PDF, TXT or read online from Scribd
    Download as pptx, pdf, or txt
    of 41

    Why use Drosophila to study

    Human disease?
    A lot already known
    • Drosophila have been studied for over 100
    years
    • Genetics experiments started in 1909 by Dr.
    Thomas Hunt Morgan
    • Discovered chromosomes as location of
    genetic content
    • Genes arranged on chromosomes
    • Recombination/Linkage
    • Spontaneous mutations

    Nobel prize: 1933


    History Continued
    • Mutagenesis techniques to study phenotypes
    induced by gene knockout
    • Molecular technique development started in
    the 1970s
    – Gene cloning
    – P-element transformation
    – Clonal analysis
    Nobel Prize 1995: Genetic Control of Early
    Embryonic Development

    • Edward B Lewis: body segments into


    specialized organs

    • Christiane Nusslein-
    Volhard and
    • Eric F Wieschaus: body
    plan and formation of
    body segments

    https://www.nobelprize.org/prizes/medicine/1995/summary/
    Genomics
    • Drosophila genome
    sequenced in 2000.
    – Human genome sequenced
    2003
    • 75% of Human disease
    genes found to have
    orthologues in Drosophila
    – Sequence identity average
    40%

    Callus & Mathey-Prevot 2002


    Genomics, continued
    • Genome sequence allowed for all sorts of new
    techniques and studies!
    – DNA binding sequences
    – RNA expression studies
    • Microarray
    • RNA-sequencing
    – Chromatin-immunoprecipitation
    – polymorphisms
    Nobel Prize 2017: molecular mechanisms
    controlling the circadian rhythm
    • Jeffrey C Hall

    • Michael Rosbach

    • Michael W. Young

    https://www.nobelprize.org/prizes/medicine/2017/summary/
    https://nobelmedia.akamaized.net
    /flashcontent/announcement_201
    7_med-interview_01_496.mp4
    Circadian Rhythms: Daily oscilations based on time of day, across all organisms
    Mutant fly with disrupted circadian clock:
    period
    • Nobel prize given for identifying which protein
    was mutated and how it functions in circadian
    rhythm
    • Period accumulates during the night and is
    degraded during the day, over a 24 hour
    period.
    PER protein inhibits per transcription
    Second circadian rhythm gene: Timeless
    Circadian Rhythm is important for human
    physiology
    Drosophila lifecycle

    http://flymove.uni-muenster.de/
    Drosophila embryo studies
    • Pattern formation
    • Cell fate determination
    • Organogenesis
    • Central and peripheral neuronal development
    • Axon pathfinding
    Time lapse video of live embryogenesis

    Fluorescently labeled nuclei

    New microscopy technique


    light sheet microscope
    low level of light to keep sample alive for a long time (possibly days)
    Imaginal Discs: Cells for adult structures are set aside in the embryo
    and grow in the larvae

    http://www.sdbonline.org/sites/fly/lewheld/id41.htm
    Drosophila larval studies
    • Developmental processes
    – Cell cycle studies
    • Physiological processes
    • Behavior
    – Foraging
    Adult structures from imaginal discs
    Drosophila metamorphosis studies
    • Imaginal disc cell proliferation
    • Imaginal disc differentiation
    • Imaginal disc organogenesis
    • Autophagy
    • Cell death
    • Hormonally regulated processes
    – 20-hydroxyecdyone (ecdysone)
    • metabolism
    Drosophila adult studies
    • Organs that function like human organs:
    – Heart
    – Lung
    – Kidney
    – Gut
    – Reproductive tract
    – Central nervous system
    • Behaviors
    – Wake and sleep circadian rhythms
    – Learning and memory
    – Feeding
    – Aggression
    – Courtship
    – Grooming
    – Response to drugs
    The Drosophila Genome
    Female karyotype Male karyotype

    Thomas C. Kaufman Genetics 2017;206:665-689

    Copyright © 2017 by the Genetics Society of America


    The Drosophila Genome
    Euchromatin:
    open chromatin
    transcriptionally active

    Heterochromatin:
    closed chromatin
    transcriptionally inactive
    structural roles
    centomere
    Polytene chromosomes
    DNA replication, but no cell division
    cycles of S and G phases, without any M phase
    up to 1024 cycles!

    Giant chromosomes

    Can see banding pattern through regular light


    microscope
    banding pattern caused by how condensed the
    chromatin in (euchromatic vs heterochromatic)
    The original polytene chromosome map drawings of Bridges (1935).

    Thomas C. Kaufman Genetics 2017;206:665-689

    Copyright © 2017 by the Genetics Society of America


    The photographic polytene chromosome maps of Lefevre (1976).

    Thomas C. Kaufman Genetics 2017;206:665-689

    Copyright © 2017 by the Genetics Society of America


    Diagrammatic representation of the extent of deletions and duplications molecularly mapped
    to the fourth chromosome.

    Thomas C. Kaufman Genetics 2017;206:665-689

    Copyright © 2017 by the Genetics Society of America


    Drosophila Genetics: Balancer Chromosomes
    • Keep sets of alleles on the same chromosome
    • Allow maintenance of heterozygous lethal
    mutations in stable stocks
    • Also useful for mutant screens
    • Don’t allow viable recombination events
    • Due to large inversions on the chromosomes
    • Elements
    • lethal recessive mutation in order to avoid
    taking over the stock
    • Dominant visible mutation to identify the
    chromosome
    Normal Recombination
    Diagrammatic representation of the paring configuration and consequences of crossing over
    in a female heterozygous for a paracentric inversion.
    Diagrammatic representation of the paring configuration and consequences of crossing over
    in a female heterozygous for a paracentric inversion.

    Thomas C. Kaufman Genetics 2017;206:665-689


    Drosophila Genome
    • 180 Megabases (Mb)
    – 120 Mb euchromatin
    – 60 Mb heterochromatin
    • 17,726 genes
    – 13,907 protein coding genes
    – 3821 noncoding genes
    • rRNA
    • tRNA
    • snRNA
    • snoRNA
    • miRNA
    • lncRNA
    • pseudogenes
    Forward Genetic Screen
    • Mutate DNA randomly
    – Ideally, only once per genome
    • Maintain mutants
    • Examine mutants for phenotype of interest
    Schematic diagram showing one potential way to screen for mutations on the autosomes.

    Thomas C. Kaufman Genetics 2017;206:665-689

    Copyright © 2017 by the Genetics Society of America


    Gal4-UAS Targeted Gene Expression System

    • Effect only specific subset of cells in vivo


    – Turn genes on, off, or label
    • Gal4:
    – Yeast transcription factor to turn on transcription
    – Not active in wild-type Drosophila
    • UAS:
    – DNA sequence that Gal4 binds to
    – Turns on transcription of anything downstream of
    the UAS sequence
    Reverse Genetics
    • Mutate specific gene and evaluate phenotype
    – Drosophila homologue of Human disease gene
    • Many mutagenesis methods
    – CRISPR/Cas9
    – Transposon mediated
    – Exise transposable element
    • Gene Silencing
    – RNAi
    Why Flies?
    • Small
    • Easy to handle
    • Inexpensive to maintain and manipulate
    • Short life span
    • Many offspring
    • External development of eggs
    • Many resources-stocks
    • Best annotated genome
    • Homologues of 75% of human disease related genes
    • Complex behaviors
    • Fewer ethical concerns
    Fly resources
    Overview and summary
    More Online Resources

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