Approaches to development:
experimental embryology
NORMAL DEVELOPMENT
- NORMAL
DEVELOPMENT
refers to the course of
development that the embryo
follows in standard laboratory
conditions
- There is no experimental
manipulations
- Development occurs naturally
- EMBRYONIC
DEVELOPMENT
is
predictable
- Mammals
and
birds:
Development occurs at a
standard
physiological
temperature
- Free-living
embryos
(Xenopus or zebrafish): rate
of development varies on
temperature
- Development
can
be
MATERNAL (due to the
components of the egg or
the genome of the mother)
or ZYGOTIC (due to the
components
newly
synthesized by the embryo
itself after fertilization)
TERMS TO REMEMBER
1. Anterior or cranial end
- front end of an animal
2. Posterior or caudal end
- rear end of an animal
3. Dorsal
- upper surface of an animal
4. Ventral
-lower surface of an animal
(chest area)
MICROSCOPIC SECTIONS
5. Transverse section
- Section taken across the
long axis of an animal
(separating the posterior and
anterior region)
6. Longitudinal
- parallel to the long axis
7. Sagittal
VERTICAL
longitudinal
section that is cut at the
midline
8. Parasagittal
VERTICAL
longitudinal
section that is cut at one side
of the midline
9. Frontal or coronal section
- HORIZONTAL longitudinal
section that separates the
dorsal and ventral sides
STAGE SERIES
- All models used for laboratory
work have published stage
series
- Describes the course of
development as a number of
standard stages (stages the
embryo goes through as it
develops) identified through
external features using a
dissecting microscope
THE FATE MAP
- Diagram that shows what will
become of each REGION of
the embryo in the course of
NORMAL development
1. Where will it move?
2. How will it change shape?
3. What structures will it
form?
- FATE MAP changes from
stage to stage due to
morphogenetic movements
and growth
MOSAIC
- Experimentally isolated parts
DEVELOPS according to the
fate map
REGULATIVE
- Isolate parts form MORE
STRUCTURES expected from
the fate map
- Depends on the double
gradient system
Ex: ADMP-Chordin system in
Xenopus
How are fate maps constructed?
1. Label SINGLE cells or regions
of embryos
2. Locate the position and shape
of the labeled patch at a later
stage of development
Labeling can be
a. Application of extracellular
label to a patch of cells
b. Injection of an intracellular
label to one cell
DEVELOPMENTAL COMMITMENT
- Formerly uncommitted cells
become committed cells to
form body parts of cell
types
- Commitment
is
the
combination of transcription
factors present in the cell so it
can be visualized directly by
observing the expression of
the relevant genes using in
situ hybridization
1. Specification
- a cell or tissue is said to be
specified
to
become
a
PARTICULAR STRUCTURE
2. Determination
- Commitment is irreversible
- It will continue to develop
autonomously even if is
moved to any other region
of the embryo
- Pathway of development is
UNALTERED
- Cells
have
lost
their
responsiveness
or
competence to the singals
that originally turned on the
relevant combination of
transcription factors
1. Orthotopic graft
- Graft to the same
position of another embryo
2. Heterotopic graft
- Graft to the different
position in the host
- Test for determination
Potency
- range of possible cells types
or structures into which a
morphogen
Ex of morphogens: sonic
hedgehod protein in the
neural tube and the limb
bud, active BMP (bone
morphogenic protein in
Xenopus)
or
Dpp
(Decapentaplegic protein)
in Drosophila
b. If the signaling center lie
in one cell sheet and
responding
cells
in
another
single
threshold response
ex: induction of nasal, lens
and otic placodes
2. Appositional induction
- only one threshold
response would be made
LATERAL INHIBITION
- Another
type
of
cell
communication
- Known in terms of the
behavior
of
Notch-Delta
system
- The individual cells from a
uniform population follow
one pathway while the
surrounding cells follow
another
Ex: neurogenesis, formation
of endocrine cells in the
epithelia of the gut
- Activator
and
inhibitor
substance
- Activator substance is short
ranged and only active
through
intercellular
contacts
- Inhibitor substance is long
ranged, moving freely with
diffusion
STOICHASTICITY IN
DEVELOPMENT
- there
is
random
determination
(random
distribution
or
random
pattern)
- DETERMINANTS
become
localized at one end of the cell
rather than the other
- Multiple cell types appear to
differentiate from a single
type of progenitor
- SYMMETRY BREAKING
THE EPIGENETIC LANDSCAPE
- Developmental commitment is
the subject of a remarkable
concept first introduced by the
embryologist and geneticist
C.H. WADDINGTON
- WADDINGTON introduced
the term EPIGENETIC as
synonym of development,
recognizing that a large
element of development
consists of the regulation of
gene expression
- The epigenetic landscape
portraying symmetry
breaking event
- Ball: cell in an embryo
- Valleys: states of
developmental commitment
- Each Bifurcation:
developmental choice
Problems
1. Diagram relates only to cell
states and does not capture
the structure of embryo
2. Exchange of inductive
signals are not represemted
3. Most developmental
choices are not symmetry
breaking events BUT
DETERMINISTIC EVENTS
controlled by inductive
signals
How is EPIGENETICS DEFINED
NOW?
- It is the understanding of gene
expression
in
terms
of
chromatin structure rather
than of DNA alone
- Importance
of
DNA
methylation as a controller of
gene
expression
and
numerous
chemical
modifications of histones and
other chromosomal proteins,
which can collectively prevent
or enable the activity of
transcription
CRITERIA FOR PROOF
1. Expression
- molecule
should
be
expressed in the right place,
right
stage
and
in
a
biologically active form
- in situ hybridization or
immunostaining
- use of retinoic acid
2. Activity
- molecule
must
have
appropriate biological activity
in a suitable test system
3. Inhibition
- molecule is in vivo (inside)
then the process for white it is
thought responsible should
fail to occur
- redundancy
- inhibition may be achieved at
the DNA level by mutation of
a gene to inactivity
- Ex:
neurogenesis,
formation of endocrine cells
in the epithelia of the gut
- At RNA level, using of
antisense morpholinos or
RNAi
- At protein level, introduction of
a specific inhibitor of the
normal gene product