March 17. Imaginal Disc Regenerative Growth

March 17. Imaginal Disc Regenerative Growth -...

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Regenerative Growth in Drosophila  Imaginal Discs Is Regulated by Wingless and Myc Miling W. , Eric S. and Jasmine C
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Regeneration Hydra Salamander
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Imaginal Discs  sheets of prospective  epidermal cells   grow during larval  development  form folded sacs of  epithelia    fate map of adult  structures
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Studies on Regeneration Previous Methods: - Surgical transplants       (Hadorn and Buck 1962) - Trans-determination experiments  (Maves and  Schubiger, 1998)
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Purpose To describe a new “unbiased” method for  identifying regulatory genes in regeneration  To further explore the role of Wg and Myc in  regeneration  (Herranz et al. 2008)
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Nonsurgical tissue-specific ablation GAL4 enhancer trap. A technique in  Drosophila  that  uses GAL4 to turn on the expression of a specific  gene in a particular tissue or stage of development GAL4 gene Encodes a transcription  factor that can turn on the  expression of any gene with  a GAL4 binding site in its  promoter region Box 2F ( Principles of Development ) 1. GAL4 gene is integrated, via a P element, into the Drosophila genome randomly 2. Under the control of a tissue-specific enhancer 3. Choose lines with GAL4 expressed only in the target tissue 3. Cross with target Drosophila that have the endogenous gene of interest with a transgenic GAL4 binding site in its promoter
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GAL4/UAS-Eiger system Eiger gene A pro-apoptotic gene that  encodes a signalling ligand  that initiates a cascade that  activates apoptosis, via  caspaces. SPATIAL control. Tissue  damage is restricted to the  larvae wing pouch
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GAL4/UAS-Eiger/GAL80 system TEMPORAL control. The timing  of apoptosis is specified. GAL80 gene Encodes a temperature- sensitive protein that acts as  an inhibitor of GAL4 At  18°C , GAL80 prevents  GAL4 from binding its  binding site on Eiger  promoter At  30°C , GAL80 does not  inhibit GAL4, allowing it to  bind to its binding site on  Eiger promoter 
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Green marks dying tissue Red marks wing pouch cells Ablation and Regenerative Growth Figure 1. Number of wing pouch cells present at A40/R0.
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Efficacy of Method The majority of the wings that underwent the  ablation-recovery process resulted in wings  75%-100% of normal non-ablated wing size Figure 2. Different sizes of adult wings after ablation recovery.
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Characteristics of Cell Regeneration 1. Localized cell proliferation 2. Quick and long-lasting
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Localized Cell Proliferation  in regenerating disc, expression of BrdU and E2F  reporter high in wing pouch and low in hinge and  notum  BrdU: thymine analogue, marks DNA synthesis  E2F: codes transcription factors that regulate cell proliferation
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This note was uploaded on 12/06/2011 for the course BIOL 430 taught by Professor Dr.ianchin-sang during the Winter '10 term at Queens University.

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March 17. Imaginal Disc Regenerative Growth -...

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