principles of experimental embryology
TRANSCRIPT
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Principles of Experimental Embryology
Biology 4361 – Developmental Biology
September 19, 2006
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Major Research Questions
How do forces outside the embryo affect its development? (Environmental Developmental Biology)
How do forces within the embryo cause the differentiation of cells?(Specification, Determination, Committment)
How do cells organize themselves into tissues and organs? (Morphogenesis and Cell Adhesion)
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The Embryonic Environment
What is the “embryonic environment”?
Internal influences (e.g. intrauterine):
- chemicals (e.g. maternal hormones, caffeine, nicotine)
- competitors (e.g. litter-mates)
Environmental regulation pathway:
- external stimulation triggers signaling event in embryo
- signal stimulates an embryonic pathway, usually hormonal, that changes the developmental pathway.
Organisms are connected to their environment
External influences:
- light
- temperature
- humidity
- predators
- competitors
- intraspecific signals
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Environmental Regulation of Development
Light, temperature & moisture = environmental signals that produce changes in embryonic hormones.
Hormones induce changes in color, reproduction, hibernation, behavior
Environmentally-produced changes in hormones during development can affect the adult phenotype; e.g.
Bicyclus anynana
seasonal polyphenism - common in butterflies
Nemoria bistriariawet season dry season wet seasondry season
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Seasonal Polyphenism
Nemoria arizonaria larvae
early spring – oak catkin (flower) summer – oak twig
Environmental signal – tannins in oak leaves (probably)
- developmental response – change in cuticle morphology
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Anthropogenic Effects
pepper moths
Industrial melanism
(“anthro” – man)
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Environmental signals – UV radiationUV = mutagenic, damaging to cell and tissue development
- lower wavelengths have more energy; cause more damage- developmental stages are the most sensitive!
melanin
Mycosporine-like amino acid = sunscreen
HO HO
NOH
COOH
NH
COOH
OCH3
R
UV exposure can induce protective mechanisms
northern leopard frog
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Bonellia viridis (marine echiuroid worm)
♂♀
Environmental Signals - Intraspecific
- larvae settles on sand/rock surface = ♀- larvae settles on proboscis = ♂
10 cm
2 mm
Therefore, signal (most likely chemical) to become ♂ is from ♀
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Environmental Signals - Temperature
- sex determined by the egg temperature during weeks 2 & 3
≤ 30° C = female
≥ 34° C = male
Temperature-dependent sex determination in alligators
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Differentiation
How does the intraembryonic environment direct cellular differentiation?
Differentiation – development of cellular specialization
Differentiation is a process
- preceded by several identifiable steps
- commitment
- specification
- determination
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Commitment
undifferentiated differentiated
specification determination
1. Specification – capable of differentiating autonomously when placed in a neutral environment.- reversible
2. Determination – capable of differentiating autonomously evenwhen placed into another region of the embryo.- essentially irreversible
Stages of cell commitment:
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Specification Types
I. Autonomous specification
II. Syncytial Specification
III. Conditional Specification
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Autonomous Specification
-Cells are specification by differential distribution of cytoplasmiccomponents during cleavage of the egg and early embryo.- proteins- RNA
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Autonomous SpecificationTunicate (sea squirt)
blastomere separation
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Autonomous SpecificationTunicate (sea squirt)
- blastomeres are committed at a very early stage in mosaic development
- each dissociated blastomerepair forms original structures
-each blastomere contains positional information in the form of specific proteins and genes
dissociated blastomeres
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Mosaic Developmentdifferentiated cell
mitosis
early embryo
later embryo
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Syncytial Specification
Syncytium – nuclear division without cell division;results in cytoplasm with many nuclei
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DrosophilaCleavage
nuclei & cytoplasm formsyncytial blastoderm
FELICE FARBER
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Syncytial Specification throughMorphogen Gradients
Drosophila egg
bicoid – anterior determinant
nanos – posterior determinant
Maternal messages:
U Irion & D St Johnson
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Syncytial Specification throughMorphogen Gradients
bicoid – anteriornanos – posterior
Maternal messages:
Each morphogenestablishes a gradientthroughout the embryo (like a diffusion gradient)
Bicoid & Nanos proteins = morphogens
1:0 10:1 1:1 1:5
- each region will have a distinct Bicoid:Nanos ratio
- Bicoid:Nanos determines anterior-posterior identity
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Syncytial Specification throughMorphogen Gradients
Cells establish identity depending on their position in multiple gradients
bicoid – anteriornanos – posterior
Maternal messages:
Each morphogenestablishes a gradientthroughout the embryo (like a diffusion gradient)
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Bicoid Protein = Head
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Bicoid Manipulation
= morphogen gradient
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Conditional Specification
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Conditional Specification
Cell fate depends on interactions with neighboring cells
Embryonic cells can change fates to compensate for missing parts = regulation
Conditional specificationproduces Regulative Development
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Conditional Specificationdifferentiated cell
mitosis
early embryo
later embryo
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Morphogen Gradients in Conditional Specification
Cells respond to protein concentration by turning different colors.
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Morphogen Gradients in Conditional Specification
Cell commitment and differentiation are programmed by various morphogen gradients.
Transplants of flag “cells”shows that they retain theiridentity (nationality), but grow according to the cellsaround them.
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Conditional Specification: Leg-Antenna Transplant
Drosophila legs and antennae are structurally-related, but obviously have different morphologically.
Consider “leg” and “antenna” to be like the national identities in the flag analogy.
Experiment: Transplant cells that would ordinarily produce the proximal (close to the body) leg to an area that would ordinarily produce the tip of the antenna.
proximal portion of legdistal portion of antenna
clawsI. Duncan
The resulting structure would be an antenna with a claw at the end.
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The transplanted leg cells have kept their “leg” identity, but have modified theirdevelopment from their original location (proximal to the body), to that of their new location (the distal-most point).
In this example of conditional specification, a morphogen gradient that started at the body (source) would specify proximal structures. As the morphogenconcentration decreased more distal structures would be formed. Therefore, while the leg cells kept their leg identity, they were “conditioned” by the very low morphogen concentration at the tip (sink) to form the most distal leg structures – claws.
I. Duncan
Conditional Specification: Leg-Antenna Transplant
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Stem Cells and Commitment
Pluripotent – makes many types of cells.
Totipotent – ability to make all cell types.
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Stem Cell Derived Blood Cells
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Morphogenesis and Cell Adhesion
How are tissues formed from populations of cells?
How are organs constructed from tissues?
How do organs form in particular locations and how do migrating cell reach their destinations?
How do organ and their cells grow, and how is their growth coordinated throughout development?
How do organs achieve polarity?
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Cell Interactions
Cell membrane protein components bind cells together; e.g.
cadherins catenins
Cells interact with each other either through paracrine signaling at some distance, or through direct contact.
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Cadherin-mediated Cell Adhesion
actinmicrofilamentsystem =anchoring andmovement
Different cellshave different cadherins.
Different cadherinshave different affinities for each other.
Thus, cell types can segregate themselves based on membrane components.
NOTE - Ca2+ -dependentbinding:
Ca2+ can control both strength andreversibility ofbinding