Whoa, Whoa, Where Did the Nerve Cells Go

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Whoa, Whoa, Where Did the Nerve Cells Go?
By Randy Hewes Research Group Faye, Danielle,
Lianne, April, Jane
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Have you or anyone you know suffered from any of
the following

• Diabetes
• Parkinsons Disease
• Alzheimers Disease
• Stroke or brain injury

If so, this research could be of interest to you
and impact your life.
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Why Bursicon Cells?

• Bursicon cells are present throughout the life
  cycle and trigger wing expansion behavior.
• Within 24 hours of wing expansion, bursicon
  cells will die.
• We can affect bursicon cells by genetically
  manipulating (turning on or off genes) within the
  bursicon cell.

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Inhibition of Insulin signaling (FOXO
overexpression) kills the Bursicon Cells
Wild Type
Yellow Mutant
Yellow Mutant
without FOXO expression
with FOXO expression
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What was the hypothesis?

• If insulin signaling is increased within cells
  that normally die, we can block cell death.

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No Fly Left Behind

• Drosophila melanoagster
• (popularly known as fruit flies).

• y w Mutant
• 2. y w Mutant with PI3K in the bursicon cells
• 3. y w Mutant with Insulin Receptor turned on in
  the bursicon cells

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The Method--Staining for bursicon
The process of immunocytochemistry

• 6. Wash
• 7. Add Secondary Antibody
• 8. Wash
• Glycerol
• Mount

• Dissection
• Fixation
• Wash and permeabilize
• Block
• Add Primary Antibody

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Dissection

• First, we had to anesthetize the fruit flies
  using carbon dioxide.

• Then, we had to remove the legs and wings.

• Next,we had to move and pin them to a petri dish.
  
• Once on the dish we had to expose the brain by
  removing the cuticle.

• Next we had to separate the cuticle of the thorax
  to expose the flight muscles.
• Then we had to remove the flight muscles to
  reveal the ventral nerve cord.

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Fixation
Wash
Block
Primary Antibody
Wash
Secondary Antibody
Wash
Glycerol
Mount
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The Results
Control
PI3K
InR
Control
PI3K
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What does it all mean?

• We expected the Bursicon Cells in the control
  group to have died.
• Although they were NOT absent, we did see a
  significant difference.
• The diameter of the PI3K cells were larger than
  the control cells and the InR cells.

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What does it all mean?

• We expected the Bursicon Cells in the control
  group to have died.
• Although they were NOT absent, we did see a
  significant difference.
• The diameter of the PI3K cells were larger than
  the control cells and the InR cells.
• There was a trend in brighter cells in the PI3K
  cells. But it was not statistically significant.

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What does it all mean?

• We expected the Bursicon Cells in the control
  group to have died.
• Although they were NOT absent, we did see a
  significant difference.
• The diameter of the PI3K cells were larger than
  the control cells and the InR cells.
• There was a trend in brighter cells in the PI3K
  cells. But it was not statistically significant.
• There was also a trend toward more cells in the
  PI3K cells. But it was not statistically
  significant.

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