Zebra Fish

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Zebra Fish

• Danio rerio

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What is a zebra fish?

• Tropical freshwater fish
• Minnow family
• Generation time is 3-4 months
• Adults 4cm in length
• Omnivorous
• Eats primarily zooplankton, phytoplankton,
  insects and larvae

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Why are zebra fish important?

• The embryos are see-through
• allows us to watch the changes in development
• Adults breed rapidly
• good supply of embryos for our research
• Embryos develop quickly
• 4-5 days old the embryos become swimming larvae
  and start to catch their own food
• Zebra Fish Development
• Close homology to humans
• More closely related to humans then models such
  as the worm and fruit fly, easier to use than
  mouse/rat

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Development
HPFHours Post Fertilization
0 HPF 1 HPF 2 HPF
4 HPF 8 HPF 12 HPF
16 HPF 24 HPF 36 HPF
48 HPF 72 HPF
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Zebra in the classroom

• Read through the zebra in the classroom article
  with a partner
• What must be done in order for zebra fish to
  successfully breed?
• Why are zebrafish embryos helpful for studying
  development?
• What does hpf stand for?
• What is it about zebra fish physiology that makes
  them good models to study in the lab?
• Explain why biochemistry and genetics are
  important aspects to understand about the zebra
  fish.
• Is it right to say that zebra fish are a
  research tool?
• What information is most important to know for
  conducting an experiment with zebra fish?

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Salt Solution (Test Salinity)
Concentration mg/Liter Stock (40.0g/Liter) I.O. Solution Distilled Water
60 .75 mL 499.25 mL
120 1.5 mL 498.5 mL
240 3.0 mL 497.0 mL
480 6.0 mL 494.0 mL
1000 12.5 mL 487.5 mL
10,000 125 mL 375.0 mL

• Must use 3 Concentrations of your choice

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pH Solution (Test acidity)
Solution Final volume of 500 mL 40 g/L Stock Solution I.O Solution Distilled Water Part 1 Additions to adjust pH to final pH
pH 4.0 0.25 mL 150 mL Add 0.1M HCl until pH 4.0
pH 7.0 0.25 mL 150 mL Add either 0.1M HCl to lower or 1M NaHCO3to raise to a final pH 7.0
pH 10.0 0.25 mL 150 mL Add 0.1M NaOH until pH 10

• Must Use 3 Concentrations of your choice
• ADD ACID TO WATER NEVER WATER TO ACID!!!

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Ethanol
Final in mM Stock in mM mL of Stock mL of Embryo Media Total volume in mL
300 1000 100 350 450
100 1000 33 300 333
30 300 33 300 333
Unstablemust make the day before use Must use 3
Concentrations of your choice
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Zebrafish Development - Zygote The Bubble

• Zygote Period 0 0.75 hpf
• Fertilized Egg to develop with its first division
  of cells

10
Cleavage Period The bubbles boil over

• Cleavage - 0.75 2.25 hpf
• Cells begin to divide still large enough to
  count

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Blastula Period The afro

• Blastula- 2.25-5.25 hpf
• Cells multiply very rapidly at the top

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Gastrula Period The Solar Eclipse

• Gastrula - 5.25- 10.33 hpf
• Outside of the cell begins to develop

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Segmentation Period The six pack abs

• Segmentation - 10.33 24.0 hpf
• Somites form around the midsection for
  segmentation.

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Pharyngula Period The fat belly

• Pharyngula - 24.0-48.0 hpf
• Fish has large yolk sack present/ Body
  straightens

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Hatching Period The free fish

• Hatching - 48.0-72.0 hpf