Vertebrate Development Flashcards

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1
Q

Fertilization, Cleavage, Gastrulation, then what?

A

Phylotypic Stage (Distinct head)

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2
Q

Phylotypic Stage

A
  1. Forms the distinct head
  2. Neural Tube is formed (place where future brain & nervous system will be
  3. Notochord- place where future spine will be, flanked by somites (which are blocks of cells for future muscle & skeleton)
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3
Q

Anamniotes

A

No amnion, fish and amphibians

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4
Q

Amniotes

A

Amnion- extra embryonic membrane (mammals & birds)

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5
Q

Frog (Xenopus)

A
  1. Have easy access to zygotes where can inject hormones to make them mate
  2. They have 100 eggs in one large blastula (which is easy to manipulate)
  3. They are hardy eggs where no infections after surgery
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6
Q

Which is better for genetic studies X. tropicalis (diploid) or X. laevis?

A

X. tropicalis which is diploid

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7
Q

Frog life cycle: The egg

A
  1. Polar where it has two region a dark region that is the animal region & a light region that is vegetal region (yolk/proteins)
  2. Stopped at meiosis - 1
  3. Fertilization- one sperm enters the animal region where it starts meiosis 2 & pronuclei fuse
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8
Q

What happens during the 1st cleavage of a frog egg?

A

1st cleavage occurs 90 mins after fertilization through animal- vegetal axis where it cuts both animal & vegetal region

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9
Q

2nd Cleavage of frog egg

A

Occurs after 30 mins where it cuts through animal- vegetal axis perpendicular to the 1st cleavage where it cuts both animal & vegetal region (all cells are equivalent)

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10
Q

3rd cleavage

A

Occurs at the equatorial axis where it separates the animal & vegetal region- 4 small animal cells & 4 large vegetal cells (both blastomeres)

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11
Q

Cleavage of the frog eggs

A

Its synchronous initially but at 4th division, not synchronous where thick vegetal yolk impedes which results in the vegetal region having fewer but larger cells

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12
Q

Blastula

A

Occurs after the 12th cleavage where it has 1,000 cells& in the animal region the blastocoel is the fluid cavity (the 3-germ layer are allocated but not moved)

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13
Q

Gastrulation

A
  1. Has a little mitosis where it moves & rearrangement of germ layers
  2. blastopore –infoldings on future dorsal side
  3. dorsal lip–organizer of AP axis structures
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14
Q

Gastrulation also involves what during the frog egg cycle?

A
  1. involution- endoderm, mesoderm occurs where they move inside through blastopore
  2. epiboly- ectoderm spreads & covers embryo
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15
Q

Neurulation (neurula)

A

Occurs after fertilization where the neutral tube precursor to CNS

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16
Q

Organogenesis

A

Involves making organs & tissues

17
Q

Describe the frog egg cycle

A
  1. Egg goes through cleavage to make blastula
  2. Blastula goes through gastrulation to make gastrula
  3. Gastrula makes neurula
  4. Neurula goes through Organogenesis to make an embryo
  5. Embryo forms tadpole
  6. Tadpole goes through metamorphosis to become an adult
18
Q

Zebrafish

A
  1. Has a short life cycle off about 3 months
  2. Embryo transparent
  3. Cleavage doesn’t cut through the yolk instead the blastomeres mound on top of the yolk
19
Q

Mutation involving zebra fish

A

lymphatic vessels grow out of control fluid accumulation in lungs (created same mutation in zebrafish fish got similar phenotypes screened for drugs saved boy’s life)

20
Q

Early studies

A

Were descriptive where they describe & draw & used microscopes

21
Q

In situ hybridization

A

They detect mRNA gene expression with labeled RNA probe where the whole embryo in situ

22
Q

Immunofluorescence

A

Detect protein gene expression with labeled antibody

23
Q

Microinjection (Used for fate mapping)

A
  1. Use for large cells
  2. Fluorescent dye
  3. GFP mRNA
24
Q

Forward genetics

A

Phenotype can be used to ID genes, it also has spontaneous mutations, which are slow rate & natural Induced mutations, which are forced & high rate (like chemicals, X-ray, UV)

25
Q

Induced mutations

A

Random mutations that can occur in every gene & can also treat large populations (breed fast)

26
Q

Zebrafish relating to mutation

A
  1. Large-scale mutagenesis
  2. high fecundity many embryos
  3. embryo transparent - large ID phenotype
27
Q

Induced mutations (with mice)

A

Dominant mutants were shown in the phenotype of F1 (F1 shows phenotype)

28
Q

Induced mutations in fish

A

Recessive mutants phenotype in F3 (F1 het. (no phenotype), F2 het fam (no phenotype), when you cross F2 fam, F3 homo recessive (phenotype)

29
Q

Knock-out

A
  1. permanent germline

2. mutate gene in ESCs in vitro

30
Q

Knock-in

A
  1. knock-out

2. replace endogenous gene (or allele) with another

31
Q

Knock-down

A
  1. block mRNA or protein not DNA

2. transient DNA intact (RNAi)