Name: Timmy Ma, Dean Long Date 3-2-16 Period 6

Investigation: What Determines Cell Differentiation in the Sea Urchin?

Procedure: Complete the Virtual Investigation (link on Blackboard). Follow the directions below and
answer the questions as you go along.

1. On the first screen - click on each of the four adult forms to see an image of the embryo that gave
rise to that adult. Complete the table below. (You can copy the embryo images from the website)
Embryo What does embryo What are the advantages/disadvantages of using the
look like? embryo in animal development studies?
Human Researchers chose to use a mouse because its
development is similar to humans. Internal
development is difficult to study with the
experimental approach
Frog Researchers are now able to raise frog embryos
through metamorphosis in labs, and experiments on
embryonic development of frogs have yielded
valuable research.
Sea Urchin Development is synchronous, while the embryos are
clearly visible and allow cell behavior to be seen
easily. Because of this, the sea urchin has been a
valuable model for researchers studying embryonic
development.
Fruit Fly The genetic approach to study genes involved in
pattern formation in development has proved
invaluable in increasing our understanding of these
processes.

Click to go to the next screen.

2. The developmental stages from zygote to larvae involves transformation stages that include :
a. Cell Division
b. Cell Migration
c. Morphogenesis

3. What are spicules?

They are structures that protrude from the ectoderm to form/support a pair of larval arms above the
mouth.
4. What are micromeres?
They will eventually give rise to mesoderm, tissue that will embryonic skeleton.
5. What are primary mesenchyme cells (or PMCs)?
PMCs separate from the epithelium and move inside.

6. What is an example of cell differentiation that has been studied extensively?

Sea urchins are usually studied thoroughly as an example of cell differentiation
7. What is the purpose of this investigation?
The purpose is to examine the differentiation of a particular cell type in sea urchin embryos. This
specific cell type will eventually form the parts of the embryonic skeleton.
8. Why will you be analyzing the position of the PMC ring and the shape of the skeletal spicules in this
investigation?
The experiment was conducted to see whether the PMC cell differentiation was regulated or
autonomous for certain genotypes using PMC transplantation experiments in the sea urchin embryo.
9. Complete the table below.
Developmental What does it look like? Major events of Stage
Stage of Sea
Urchin
Fertilized Egg Divisions of the cleavage stages occur in protective
membrane 4 large macromeres, 4 small
micromeres.
16 cell stage Mesomeres give rise to ectoderm. Half of
macromeres give rise to ectoederm, all of
endoderm, and some of mesoderm. Form blastula.

Blastula Cells separate from epithelium and move inside, that

will eventually form embryonic skeleton.

Mesenchyme Dimple forms on bottom and will extend upwards as

blastula a tube to become digestive tract. First skeletal
spicules will form.

Middle/late Mouth forms, branches extend to form larval arms

gastrula and bottom of embryo.

Prism stage Lower pair of larval arms form, mouth completed,

arms will elongate to form pluteus.

Pluteus larva Arms and mouth have grown, digestive tract

develops into esophagus, stomach, and intestine,
pluteus begins to float and feed.
When you are ready to perform the first set of experiments, continue to the next step.

10. As you go through the next 3 screens performing tests; complete the data table below.

Host
Species A Species B
Species A RP: Bottom, same as both donor and RP: Top, same host species
host (both the same species)

SF: Simple, same as both donor

SF: Simple, same as both donor and
host (both the same species)
Donor
Species B RP: Bottom, same donor species
species.
RP: Top, same as both donor and host
(both the same species).

SF: Fenestrated, same as donor

species SF: Fenestrated, same as both donor
and host (both the same species).

RP = Ring position
SF = Spicule form

11. What is the purpose of transplanting PMCs from a species A donor embryo back into a species A
host embryo (or transplanting PMCs from a species B donor embryo back into a species B host
embryo)?
Transplanting from species A donor embryo to the species A host embryo allows us to determine
where the location ring is in the spicules. If the same species is used to donate and host, then the
ring position will be the same, and eliminates outside factors on the ring shape.

12. Analyzing the data from all four experiments in the first set involving PMC ring position, what can
you conclude? Is the ring position determined autonomously, by the PMCs themselves, or regulated
by other cells? If the latter, which cells? (Note, to answer these questions, it may help to go back to
the data table and classify the ring position observed in each experiment as either "donor" or "host",
if you haven't already done so.)

From the data, it can be seen that the host cells determine the ring position. If it were autonomously
determined, then there would be no pattern with the ring shape.

13. Analyzing the data from all four experiments in the second set involving spicule form, what can you
conclude? Is the form of the spicule determined autonomously, by the PMCs themselves, or is it
 regulated by other cells? If the latter, which cells? (Note: Again, it will help to classify the spicule
form data as either "donor" or "host" in each experiment.)
It can be concluded that the form of spicule is determined by the donor cells. This can be seen when
the shape the spicule took on that of the donor cells, not the hosts. Autonomously determined
would have results with no sort of pattern to the determination of spicule form. This would be
randomly determined with no sense of relation to the host or donor; it would be completely
arbitrary. Since this pattern was not arbitrary, it could not have been autonomously.

14. Considering all the data from both sets of experiments, can you make a blanket statement about
whether PMC fate and cell differentiation is determined autonomously, or is regulated by other
cells? If not, what can you conclude?
The fate of an individual cell depends upon internal pre programming rather than on its external
environment. Cell to Cell interactions are necessary to determine whether a particular
developmental decision is autonomous or not.

Click on the "Prediction" button at the bottom of the diagram on screen 5 to make a prediction based on
your model.

15. Given what you postulated, what would you predict about the cells in the dish? Would they form a
ring, and if so, where? What form will the spicules have?
Yes because even if the micromeres are removed and there are no embryonic cells present they will
continue to divide, migrate, and form spicules under some conditions.

Click to go to screen 6.

16. Read and summarize the information on this screen as a conclusion to this investigation.

Because of developmental biology scientists have been able to study cell differentiation and how it
leads the pathway from a single celled organism to a large multicellular and complex organism we
see today.