FATE MAPS

Course: B.Sc.(H) Zoology VI semester

Paper: Developmental Biology
Faculty: Dr. Priya Goel
 FATE MAPS
• Fate map is a diagrammatic representation of the prospective fate
of each part of an embryo at an early stage of development
• Embryonic regions with a distinct fate are called Primordia /
Rudiments
• Fate maps change over time –as- cells multiply & move relative to
each other
• Thus, fate maps indicate fates at a closely following stage
• A series of fate maps at consecutive stages shows the progression
of different cells or regions through longer periods of development
• Significance:
– Essential tools in most embryological experiments
– Help to trace cell lineage by following the fate of parts of early embryo
during development*
– Helps understand the mechanism of morphogenetic movements
during gastrulation
 Different ways of establishing fate
maps
• Observing living embryos with few cells - under the
microscope – augmented with histological sections &
time-lapse motion pictures
• Large/opaque embryos can be mapped by labeling specific
cells
• Mammalian embryos are among the most difficult to
map (since they develop inside another organism), and
researchers are actively constructing, refining, and
arguing about the fate maps of mammalian embryos.
• Characteristics of a good label
– should not spread to neighbouring cells
– readily detectable at later stages
– should not interfere with normal cell development
LABELING METHODS
NATURAL MARKERS
– natural colour differences in the cytoplasm of early blastomeres
– fertilized eggs of Ascidians e.g. Ciona, Tunicate (Sea squirt) Styela
partita (Cynthia)
– E.G.Conklin (1905): followed the fate of each blastomere
• cells containing clear cytoplasm  ectoderm
• Dark grey yolky  endoderm
• Light grey  notochord & neural plate
• Yellow  mesoderm
ARTIFICIAL METHODS
• VITAL DYES: Vogt (1925)
– stain cells but do not kill them: Nile Blue Sulphate, Neutral red,
Bismark brown, Janus green
– spread dye mixed with agar / cellophane on the slide  dried 
pressed against a chosen area of blastula for a short period  stain
diffuses to blastomeres  stained cells’ movements are followed
within the embryo
– Several areas can be marked separately simultaneously & their fate
traced
– But become diluted with each cell division

• CARBON PARTICLES:
– N. Spratt (1946) – fate map of chick: C-particles stick on the cell
surface & used as markers to trace the cell fate
– William W. Ballard (1981) – modified technique: injected C-particles
or chalk particles inside the particular region of teleost embryo for
tracing its fate
– But become diluted with each cell division
• RADIOACTIVE MARKERS: C14, P32, H3 in Chick embryo
– Cells with these markers are studied by autoradiography to trace
their fate
– Tritiated thymidine labels the nuclei when incorporated into the DNA
of cells
– A region of interest is cut from host embryo  replaced by
radioactive graft from donor embryo
– Limitation: become diluted with each cell division; exposure to
radioactivity
• HISTOCHEMICAL STAINS: Enzyme specific staining of embryonic cells
 can be visualized by adding appropriate substrate for its enz.activity
– e.g. enz. Horseradish peroxidase (HRP)
• FLUORESCENT DYES:
– fluorescent dyes conjugated to large, metabolically inert carrier molecules, that
will not cross cell membranes or pass through gap junctions  microinjected into
one or more cell, all descendants of the injected cell are labeled distinctly under a
fluorescent microscope
– e.g. fluorescently labeled DEXTRAN: Fluorescein Dextran Amine (FDA) &
Rhodamine Dextran Amine (RDA)
– e.g. fluorescent carbocyanine dyes: dialkyl indocarbocyanine (DiI) & dialkyl
oxocarbocyanine (DiO) are lipophilic membrane stains that diffuse in the cell 
produce red & green emissions resp.
– Microinjection of stable mRNAs encoding fluorescent proteins: long lasting label
• GENETIC MARKERS
– One way of permanently marking cells and following
their fates is to create embryos in which the same
organism contains cells with different genetic
constitutions.
– Advantages: donot spread to neighboring cells, If
stably expressed, are inherited by the descendants of
the marked cell
– Limitations: Low efficiency of introducing the gene in
the cell
– e.g. used to create Chimeric Embryos: by Xenoplastic
transplantation of embryonic grafts from animal of
interest having different genetic constitution but
similar development pattern
e.g. Chick-quail chimeras are made by grafting embryonic quail cells inside
a chick embryo while the chick is still in the egg.
• Chicks and quail embryos develop in a similar manner (especially during the
early stages), and the grafted quail cells become integrated into the chick
embryo and participate in the construction of the various organs
• The chick that hatches will have quail cells in particular sites, depending on
where the graft was placed.

Quail cells also differ from chick cells in

several important ways, including
the species-specific proteins that
form the immune system.
There are quail-specific proteins can be
used to find individual quail cells,
even when they are “hidden”
within a large population of chick
cells
By seeing where these cells migrate,
fine-structure maps of the chick
brain and skeletal system have been
produced
 Chimeras dramatically confirmed the extensive migrations of
the neural crest cells during vertebrate development

• Mary Rawles (1940) showed that the pigment cells (melanocytes) of the
chick originate in the neural crest, a transient band of cells that joins the
neural tube to the epidermis.
– When she transplanted small regions of neural crest-containing tissue
from a pigmented strain of chickens into a similar position in an
embryo from an unpigmented strain of chickens, the migrating
pigment cells entered the epidermis and later entered the feathers

Chick resulting from transplantation of a trunk neural crest region

from an embryo of a pigmented strain of chickens into the same
region of an embryo of an unpigmented strain. The neural crest
cells that gave rise to the pigment migrated into the wing epidermis
and feathers
• Ris (1941) used similar techniques to show that, although almost all of
the external pigment of the chick embryo came from the migrating
neural crest cells
– the pigment of the retina formed in the retina itself and was not
dependent on migrating neural crest cells.
– This pattern was confirmed in the chick-quail chimeras, in which the
quail neural crest cells produced their own pigment and pattern in
the chick feathers.
Transgenic DNA chimeras
• most animals - difficult to meld a chimera from two species.
• Hence, better to transplant cells from a genetically modified organism
(retrovirus)  the genetic modification can then be traced only to those
cells that express it
• Retrovirus marking: incorporating retrovirus engineered reporter gene
into DNA of host cells  expression of reporter gene 
histochemical/fluorescent marking of gene products
• Expression of host DNA to express Green Fluorescent Protein (GFP)
– naturally occuring in some jellyfishes
– When the infected embryonic cells are transplanted into a wild-type
host, only the donor cells will express GFP
• BRAINBOW: a recent technique used to tag neurons in brain
– fluorescent proteins are used to tag different proteins in the cell  an
array of colours are generated by different expression of distinct
fluorescent proteins
Experiment: Fate mapping with transgenic DNA
showed that the neural crest is critical in making the
gut neurons
• Freem et al (2012) used transgenic techniques to study the
migration of neural crest cells to the gut of chick embryos, where
they form the neurons that coordinate peristalsis—the muscular
contractions of the gut necessary to eliminate solid waste.
• The parents of the GFP-labeled chick embryo were infected with a
replication-deficient virus that carried an active gene for GFP
• This virus was inherited by the chick embryo and expressed in every
cell  glowed green when placed under ultraviolet light
• They then transplanted the neural tube and neural crest of a GFP-
transgenic embryo into a similar region of a normal chick embryo
• A day later, they could see GFP-labeled cells migrating into the
stomach region and by 7 days, the entire gut showed GFP staining
up to the anterior region of the hindgut
 FATE MAPS OF VERTEBRATES AT
THE EARLY GASTRULA STAGE:
All are dorsal surface views
(looking “down” on the embryo at
what will become its back).
Despite the different appearances
of the adult animals, fate maps of
these four vertebrates show
numerous similarities among the
embryos. The cells that will form
the notochord occupy a central
dorsal position, while the
precursors of the neural system lie
immediately anterior to it. The
neural ectoderm is surrounded by
less dorsal ectoderm, which will
form the epidermis of the skin. A
indicates the anterior end of the
embryo, P the posterior end. The
dashed green lines indicate the
site of ingression—the path cells
will follow as they migrate from
the exterior to the interior of the
embryo.
 EXAMPLES (DIY)
• Fate map of Amphioxus
• Fate map of frog Xenopus laevis
• Fate map of chick
• Fate map of mouse