Objectives:

To understand what is Fate Mapping and

its history.
To understand the cell fate, the
commitment and the mechanism of
developmental commitment involved.
To know the different techniques used in
Fate Mapping and its different processes.

History & Proponents:
1880 The first fate maps were made.
1905 The first comprehensive collection
of Ascidian (sea squirt) fate maps was
published by Edwin Conklin.
1929 Walter Vogts research, the
amphibian blastula, divides into three
regions: animal, marginal, and vegetal.
Each of these areas houses progenitors of
the cells that will make up the future
organs of the organism.

1969 Nicole Le Douarin pioneered the
use of chickquail chimaeras for fate
mapping.
1971 More detailed fate maps have
been created for the frog Xenopus, such
as the one published by Osamu Nakamura
and Keiko Kishiyama.
History & Proponents
1974 Several experiments were initiated by
Sydney Brenner, a biologist and 2002 Nobel
Prize winner in Physiology or Medicine.
These he chose the nematode worm for
study because of its rapid period of
embryogenesis and very few cell types.
Balinskys An Introduction to
Embryology (1981) showed an image of the
fate map of the amphibians
Discoglossus and Ambystoma similar to
those created by Vogt for Xenopus.
History & Proponents
More recent illustrations, in Hake and
Wilts Principles of Developmental Biology
(2004) showed how a fate map can be
made using an amphibian egg.
Scott Gilberts Developmental Biology
(2006) showed fate maps for several
different model organisms, including the
zebra fish, frog, mouse,
and chick embryos.

History & Proponents
Cell Fate & Commitment
Developmental Hierarchy
The number of different cell types in
the embryo increases as
development proceeds, but new
cell types arise from particular pre-
existing cell types through
hierarchical series of decisions.
Cell Fate & Commitment
Cell Fate and Potency
Cell fate describes the range of cell
types a particular cell can give rise to
during normal development, while cell
potency describes the range of cell
types that a cell can give rise to in all
possible environments.
Cell Fate & Commitment
Fate Maps
A fate map is a map of an embryo
showing which parts of the embryo
give rise to a particular adult tissues.
Fate maps are generated by marking
or labeling cells in the early embryo
and following their progress through
development.
Cell Fate & Commitment
Levels of Developmental Commitment
In animals, cells become committed to
certain fates in stages, first reversibly and
then irreversibly. A nave cell may receive
information in the form of cytoplasmic
determinants or inductive signals that
specifies its fate, but this fate can be altered
by placing the cell in a different
environment. A cell becomes determined
when its fate is irreversible. This coincides
with the loss of competence to follow
alternative developmental pathways.
Mechanisms of Developmental
Commitment
How Cells Differentiate?
To differentiate, cells must begin to
synthesize new proteins. This requires the
selective expression or activation of
regulatory molecules such as transcription
factors that control which proteins are
synthesized in the cell.
Mechanisms of Developmental
Commitment
Cytoplasmic Determinants
Are molecules in the cytoplasm of a cell
that help to determine cell fate.
Induction
Is a process whereby one cell or group of
cells can influence the developmental
fate of another, and is a common strategy
to control differentiation and pattern
formation in development.
Mechanisms of Developmental
Commitment
COMPETENCE
Induction relies both on ability of
the including cell to produce the
signal and the ability of the
responding cell to receive the signal
and react in the appropriate
manner.
Mechanisms of Developmental
Commitment
Instructive and Permissive Induction
Instructive induction occurs when the
responding cell has a choice of fates, and the
inductive signal instructs the cell to adopt one
of those fates in preference to the other.
Permissive induction occurs when the
responding cell is already committed to a
certain developmental pathway, but needs
the inductive signal to continue towards
differentiation.
Mechanisms of Developmental
Commitment
Lateral Inhibition and the Community
Effect
In lateral inhibition, differentiated cells
arise in a regularly spaced pattern within
a group of equivalent undifferentiated
cells due to random fluctuations in levels
of a ubiquitous signal that inhibits
differentiation.

Mechanisms of Developmental
Commitment
In the community effect, populations of cells
can change their collective fate by
secreting enough of an inductive signal to
reach a critical concentration threshold,
whereas isolated cells follow a different
developmental pathway because they
cannot produce enough of the signal on
their own.
Different Fate Mapping Techniques
In 1905 Edwin G. Conklin conducted
the first cell lineage experiments, which
involved following the progenitor cells
of the embryo of the tunicate Styela
partita.

In 1929 Walter Vogt invented a
process in which vital dye and agar
chips are used to stain a specific
region of a developing amphibian
embryo. To do this, Vogt spread dye
and agar on a microscope plate and
allowed it to dry. He then cut small
pieces of the dried agar and applied
it to a desired part of the embryo.
Radioactive labeling and fluorescent
dyes are both relatively simple
experimental tools that use a donor
and a host embryo to follow cell
migration. The donor embryo is
treated with dye or irradiated and a
graft from the donor is removed and
placed onto the host embryo where it
joins the developmental process.
Another process for fate mapping was invented by
Nicole Le Douarin, a developmental biologist who
created chimeras, or animals with two or more sets
of genetically distinct cells. Le Douarin removed a
portion of neural tube and neural crest from a
chick embryo and replaced it with an identical
portion of neural tube and neural crest from a quail
embryo at the same stage of development. Le
Douarin also discovered that Feulgen stain
distinguishes quail cells from chick cells, which
allowed her to trace the migration of quail cells.
Genetic fate mapping (GFM) uses
two genetically engineered alleles,
one of which expresses a site-specific
recombinase, such as cyclization
(Cre) or flipase (Flp), while the other
contains a reporter allele such as
green fluorescent protein (GFP).
Utero fate mapping
A cell is grafted from a
donor animals and dyed and
was injected into the embryo
inside the uterus.



Inducible fate mapping (GIFM). This
technique generates the Cre fusion
proteins used in GFM with a
tamoxifen-responsive estrogen
receptor ligand binding domain
(CreER). CreER is removed from the
cytoplasm of the cell via heat shock
protein 90 (Hsp90).
Introduction
Cell migration is an important determinant of
brain structure. Most, if not all, nerve cells have to
migrate from the sites where they are born to places
where they terminally differentiate and integrate into
the brain circuitry. Radial migration of young neurons
from germinal regions lining lateral ventricles to
more superficial layers of the neocortex has been
recognized as a prerequisite for proper
morphogenesis and function of the cerebral cortex
(Caviness and Rakic, 1978; Sidman and Rakic, 1973; Walsh and Goffinet,
2000).
Introduction
Radial migration could explain the radial
organization of the cerebral cortex and the
inside-out sequence of cortical formation, and
as such it has been for many years considered
to be the principal mode of cell translocation in
the developing cortex (Rakic, 1988). This view
began to change when retroviral lineage
tracings revealed that a large number of
neurons do not follow the radial path, but
disperse tangentially throughout the developing
neocortex (Price and Thurlow, 1988; ORourke, 1992; Walsh and
Cepko, 1992).
It has been proposed that tangentially and radially
migrating neurons belong to different cell pools that
segregate early in the embryonic development (Tan
and Breen, 1993; Tan et al., 1998). Finally, several
experiments suggested that many of the tangentially
migrating neurons are not born in the cortical
neuroepithelium as was always assumed, but
instead originate outside the neocortex in the basal
forebrain in regions called ganglionic eminences
(reviewed by Anderson et al., 1999; Parnavelas, 2000).
Introduction
From these findings emerged a new concept of
cortical development, which proposes that two
separate populations of neurons participate in
corticogenesis. One population consists of the
radially migrating neurons, which are born in
neocortical ventricular zone and probably give
rise to the principal pyramidal neurons of the
neocortex. The second population includes
neurons born in ganglionic eminences that
migrate tangentially into the neocortex and
probably differentiate into the cortical non-
pyramidal neurons (Anderson et al., 1999; Parnavelas, 2000).

Introduction
Materials and Methods
Donor cell preparation
Ventricular and subventricular zones
from ganglionic eminences were dissected
from 5-15 E13.5 mouse embryos as
described previously (Wichterle et al., 1999).
Swiss-Webster (Taconic), CD-1 (Charles River) or
transgenic mice expressing human placental
alkaline phosphatase in all cells (DePrimo et al.,
1996) were used as donor animals.

Materials and Methods
Transplantation in utero
High density cell suspension (~800,000 cells/ml)
was front-loaded into beveled glass micropipettes (~50 mm
diameter) that were prefilled with mineral oil and mounted
on a microinjector (modified version of Narishige). Cells
were allowed to settle inside the pipette and the excess
cell-free medium was expelled. The recipient pregnant
mice (E13.5) were anesthetized, uterine horns were
exposed through an intraperitoneal incision and animals
were mounted under the biomicroscope as described
previously (Liu et al., 1998; Olsson et al., 1997). The tip of
the micropipette was inserted into the LGE or MGE under
real-time ultrasound guidance and 10-20 nl of cell
suspension was injected.
Analysis of cell migration
Transplanted embryos (~60% of
injected embryos survived) were collected
at 1, 2 and 4 days after transplantation or as
3-week and 4-month-old animals. Three to
six successfully transplanted animals were
analyzed for each time-point and region
(~50% of surviving injected animals
contained grafted cells).
Materials and Methods
Fig. 1.
Distribution of MGE
cells one (E14.5, A-C),
two (E15.5, D-F) and
four (E17.5, G-I) days
after homotopic
transplantation.
Coronal
sections of embryonic
mouse brains
containing MGE cells
labeled with PKH26
fluorescent dye before
transplantation. Graft-
derived cells are black
or dark blue.

Fig. 2.
MGE cells in the
subventricular and
marginal zones at
two (E15.5) and
four (E17.5) days
after
transplantation.
Fig. 3. MGE cells expressing alkaline
phosphatase were grafted into the wildtype
E13.5 MGE.
Fig. 4. Morphology of MGE-derived neurons stained for alkaline
phosphatase.
Fig. 5.
Immunohistochemical
characterization
of grafted MGE (A-C)
and LGE (D-F) cells in
the neocortex (NCx;
A,B,F) and striatum
(Str; C,D,E).
Fig. 6. Homotopic
transplants
of PKH26-labeled
LGE cells.
Fig. 7. Alkaline
phosphatase-
expressing
LGE cells in 3-week-
old animals.
(B,D,E) Two to three
fused photographs
taken in different
focal planes.
Discussion
Summary
Recent studies suggest that neurons
born in the developing basal forebrain
migrate long distances perpendicularly to
radial glia and that many of these cells
reach the developing neocortex. This
form of tangential migration, however,
has not been demonstrated in vivo, and
the sites of origin, pathways of migration
and final destinations of these neurons in
the postnatal brain are not fully
understood
Summary
Using ultrasound-guided transplantation
in utero, we have mapped the migratory
pathways and fates of cells born in the
lateral and medial ganglionic eminences
(LGE and MGE) in 13.5-day-old mouse
embryos. We demonstrate that LGE and
MGE cells migrate along different routes
to populate distinct regions in the
developing brain...
We show that LGE cells migrate
ventrally and anteriorly, and give rise to
the projecting medium spiny neurons in
the striatum, nucleus accumbens and
olfactory tubercle, and to granule and
periglomerular cells in the olfactory bulb.
By contrast, we show that the MGE is a
major source of neurons migrating
dorsally and invading the developing
neocortex
Summary
MGE cells migrate into the neocortex
via the neocortical subventricular zone
and differentiate into the transient
subpial granule neurons in the marginal
zone and into a stable population of
GABA-, parvalbumin- or somatostatin-
expressing interneurons throughout the
cortical plate.
Summary
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