E-55, Exam 2-2011: Name Page 1 of 9

BIOL E-55 Developmental Biology

Exam 2 2011

A. Be sure to write your name on EACH PAGE of the examination.

B. Read each question carefully and think before you write.

If your answer cannot be read, it cannot be graded.

F.
C. If you do not show all of your reasoning, we cannot assign partial credit.
BUT BE CONCISE!

IF YOUR ANSWER HAS INCORRECT INFORMATION IN IT, POINTS WILL BE

DEDUCTED!!!

D. The point value of each problem is noted. Allocate your time accordingly.

IF YOU WRITE IN PENCIL YOU CANNOT ASK FOR A REGRADE

S.
Relax and do well!! Good luck!

page 2______ (14 pts)

page 3______ (14 pts)

L.
page 4______ (14 pts)

page 5______ (8 pts)

page 6______ (20 pts)

page 7______ (20 pts)

page 8______ (10 pts)

TOTAL /100 POINTS

E-55, Exam 2-2011: Name Page 2 of 9

Multiple Choice (2 points each). Select the best answer from the choices provided.

______1. In the sperm, the function of the acrosomal vesicle is:

A. to form the centrosome of the zygote
B. to provide enzymes that digest the outer coat of the egg
C. to contain mitochondria that power sperm movement
D. to provide proteins necessary for sperm movement
E. all of the above

______2. Which of the following are not necessary for movement of the sperm flagellum:
A. dynein
B. microtubules
C. mitochondria
D. centriole/centrosome

F.
E. filopodia

______3. The main purpose of gametogenesis is to:

A. make a haploid cell
B. specialize the gametes
C. organize the 3 germ layers
D. A and B
E. B and C

______4. In the egg, _________ contain(s) proteolytic enzymes that are released by exocytosis
after fertilization and are important for preventing polyspermy
S. A. cortical granules
B. vitelline envelope
C. acrosome
D. jelly layer
E. centrosome

______5. Which of the following is an example of a factor that helps to achieve species specificity
in sea urchin fertilization?
A. actin
B. microfilaments
C. bindin
D. cortical granules
L.
E. centrosome

______6. Which is NOT a property of resact?

A. influences flagellar movement
B. gives directional cues
C. confers species specificity
D. degrades jelly layer
E. acts as a chemokinetic

______7. Fate mapping in a frog shows that cells in a specific region of a blastula stage embryo
always give rise to the blood cells. These cells in the blastula embryo are best described as:
A. specified
B. committed
C. determined
D. differentiated
E. none of the above
E-55, Exam 2-2011: Name Page 3 of 9

______8. A specific chemical can prevent the degradation of the mitosis promoting factor (MPF).
What results if this chemical were added to early cleavage stage frog embryos?
A. the cells cannot exit mitosis and enter S phase
B. the cells cannot exit S phase and enter mitosis
C. the cell enters mid blastula transition
D. the cell begins zygotic transcription
E. none of the above

______9. The first step in sea urchin gastrulation is

A. the ingression of the primary mesenchyme into the blastocoel
B. the invagination of the primary mesenchyme into the blastocoel
C. the convergent extension of the primary mesenchyme into the blastocoel
D. the delamination of the vegetal plate into the blastocoel
E. the involution of the ectoderm into the blastocoel

F.
______10. During sea urchin gastrulation,
A. the cells that will form the ectoderm move inside the embryo
B. the cells that will form the endoderm and mesoderm move inside the embryo
C. the cells that will form the endoderm and mesoderm move outside the embryo
D. the cells that will form the ectoderm and mesoderm move inside the embryo
E. the cells that will form the archenteron move outside the embryo

______11. You have a female fly that has mutations in both copies of the gurken gene. In her
ovary, her cells cannot produce functional gurken protein. What phenotype might you reasonably
expect to obtain in her progeny?
S.
A. the embryos will develop normally
B. the embryos will be dorsalized
C. the embryos will be ventralized
D. the oocytes will be infertile
E. half the embryos will be normal, and half will be dorsalized

______12. The following are important for setting up the anterior-posterior body axis in Drosphila
A. bicoid and pipe
B. bicoid and nanos
C. cactus and nanos
D. cactus and pipe
E. none of the above
L.
______13. Frog embryos are injected with a morpholino against fibronectin. Which of the following
developmental processes will be affected:
A. neural tube closure
B. migration of cells during gastrulation
C. the first cell division will be blocked
D. the first nuclear division will be blocked
E. branching morphogenesis of the metanephric kidney

______14. Dorsal endoderm with is cultured with ectoderm. The ectoderm is converted to:
A. ventral ectoderm
B. ventral mesoderm
C. ventral endoderm
D. organizer
E. Nieuwkoop Center
E-55, Exam 2-2011: Name Page 4 of 9

______15. These 3 signaling pathways are important for induction and formation of the organizer:
A. TGF-β, Wnt, Nodal
B. Notch, Wnt, Nodal
C. BMP, Fibronectin, Nodal
D. TGF-β, FGF, Nodal
E. TGF-β, Notch, Nodal

______16. In a mammalian embryo, the trophectoderm

A. gives rise to the inner cell mass
B. gives rise to the embryo
C. gives rise to extraembryonic tissues
D. is pluripotent
E. all of the above

F.
______17. The neural tube becomes separated from the epidermis by
A. expression of N-cadherin in the epidermis
B. differential cadherin expression in epidermis versus neural plate
C. induction of medial hinge point cells by the notochord
D. pushing forces exerted by the migrating neural crest
E. wedge-shape morphological changes in the medial hinge point cells

______18. Which pathway(s) relies on activated smads to transduce its signals?

A. Notch signaling
B. TGF-β signaling
C. Wnt signaling
S.
D. Hox gene signaling
E. both B and C

______19. Which of the following is not derived from the ectoderm

A. neural tube
B. spinal cord
C. epidermis
D. muscle
E. pigment cells

______20. Determination of dorsal ectoderm to become neural rather than epidermal requires:
A. BMP signals from the organizer
L.
B. BMP antagonist signals from the organizer
C. BMP antagonist signals from the ventral mesoderm
D. BMP signals from the ventral mesoderm
E. maternally supplied Nodal signals

______21. Injection of Noggin mRNA in cells that will become the future ventral side of a frog
embryo mimics the effect of an organizer graft to the ventral side. This experiment demonstrates
A. that Noggin is a transcription factor
B. that Noggin induces ventral fates
C. that Noggin is committed to organizer fates
D. that Noggin is required to induce a secondary axis
E. that Noggin is sufficient to induce a secondary axis
E-55, Exam 2-2011: Name Page 5 of 9

______22. The neural crest is a population of cells that

A. form at the border between the epidermis and neural tube
B. undergo an epithelial to mesenchymal transition
C. migrate to distant parts in the body
D. none of the above
E. all of the above

______23. After removing the epidermis, the neural tube:

A. lacks ventral neural fates
B. lacks dorsal neural fates
C. an ectopic roofplate is induced
D. more sensory relay neurons are formed
E. none of the above

F.
______24. Removing the epidermis from a developing embryo and observing the effect on the
neural tube, is a
A. loss-of-function experiment
B. gain-of-function experiment
C. gene expression experiment
D. fate map experiment
E. knock-down experiment

______25. Fate mapping in a 16-cell stage sea urchin embryo shows that the mesomere cells,
which are in the animal pole, always give rise to the ectoderm. You then perform a series of
experiments. In the first experiment, you remove the mesomeres, culture them alone in a dish,
S.
and see that they still give rise to ectoderm. In the next experiment, you remove the mesomeres
and transplant them adjacent to the vegetal pole of another 16-cell stage embryo (in a region that
will become endoderm). This time, your transplanted cells develop into the endoderm. From these
experiments, you can conclude that the mesomere cells at the 16-cell stage are:
A. fated to become the ectoderm, but their state of commitment is not known
B. specified, but not determined to become ectoderm
C. specified but not determined to become endoderm
D. determined but not specified to become ectoderm
E. both specified and determined to become ectoderm
L.
E-55, Exam 2-2011: Name Page 6 of 9

Matching (10 points). Match the letter that best describes the following words:
(note: not all answers are used!)

26. __K___resact
27. __H___bindin
28. __A___zona pellucida
29. __D___cytokinesis
30. __F___ingression
31. __I___delamination
32. __J___in situ hybridization
33. __C___immunohistochemistry
34. __E___targeted gene knock-out
35. __G___RNAi

F.
A. a protective layer of glycoproteins, analogous to the vitelline envelope
B. mitotic division of the nucleus by the mitotic spindle
C. a technique that detects protein expression of a specific gene
D. division of the cytoplasm by a contractile ring of microfilaments
E. a technique that can be used for loss-of-function studies in mouse
F. migration of individual cells into an embryo
G. a technique that can be used for loss-of-function studies in frogs and drosophila
H. a protein that ensures species specific binding between sperm membrane and vitelline envelope
I. splitting of one epithelial sheet of cells into two sheets
J. a technique that detects mRNA expression of a specific gene
K. a species specific chemotaxic compound that attracts sperm
S.
Short answer (point values indicated). Provide a concise response in the space provided.

36. (10 points) Define the following terms and describe one example for each (such as
one from those we have discussed in class about gastrulation movements in sea urchin
or frogs).

A. convergent extension
L.
The cells merge together or intercalate to narrow and then extend the tissue (2.5 points).
Example: This occurs in the formation of the archenteron in sear urchins where the gut tube
changes from a wide and short gut rudiment to a long thin tube.
This also occurs during frog gastrulation: (1) after involuting through the blastopore, the
mesoderm intercalates along the mediolateral axis, forming a long narrow band or (2) the
ectoderm is undergoing radial intercalation to make a thin band of flattened cells or (3) in chick
neurulation, before the induction of the medial hinge point cells, the neural plate lengthens and
narrows by convergent extension.
(2.5 points for one example)

B. epiboly
It is characterized by the movement of an epithelial sheet that spreads and encloses the deeper
layers of an embryo. It can occur by cell division, cell shape changes, or by cells intercalating into
fewer layers. (2.5 points definition)
Example: In frogs, the animal cap and noninvoluting marginal zone cells expand by epiboly to
cover the entire embryo and will form the ectoderm. (2.5 points for one example)
E-55, Exam 2-2011: Name Page 7 of 9

37. (10 points) A new cadherin gene is isolated from zebrafish ectoderm at the end of
gastrulation. Knockdown of this cadherin with morpholinos results in normal brain and
spinal cord development, and normal epidermis. Interestingly, researchers notice that
the jaw structures in the morpholino-injected embryos look abnormal, and they also
observe that the heartbeat is irregular, with blood flow in the cardiac outflow tract
appearing to be restricted.

A. Based on these data, propose a hypothesis as to the role of this gene in ectoderm
development.

This was a question on sample EXAM B on the website!

Hypothesis: the novel cadherin mitigates neural crest cell formation. The diverse defects observed
in cartilage and heart vessel development could be explained by a common deficiency in the ability
of neural crest cells to form/migrate properly. One could speculate that this cadherin makes the

F.
neural crest cells gain affinity for one another or lose affinity (to detach from) neighbors in the
forming epidermis and neural plate/tube. (4 points)

B. Name one piece of data that would provide correlative support for your hypothesis.

Look for gene/protein expression of the cadherin specifically in wild-type neural crest cells (in situ
hybridization, immunohistochemistry, isolate neural crest cells and perform RT-PCR are among the
possible). A new technique must be described. Listing of the above the observations was not
accepted (3 points)

C. Name one piece of data that would provide functional support for your hypothesis.
S.
Look in morpholino-injected animals at the neural crest: observe whether is it formed, does it fail
to migrate. Alternative answers involve specifically knocking-down or rescuing cadherin expression
in the neural crest cells (3 points). The answer must include some type of functional assay (a loss-
of-function or gain-of-function technique) to receive credit.

38. (10 points) In 2-3 sentences, summarize how the organizer is formed: what
signal(s) contribute to organizer formation, and where does this occur in the amphibian
blastula? You *may* use a diagram to explain your answer.
L.
Following sperm entry, cortical rotation causes the displacement of Disheveled and GSK3-binding
protein (GBP) to the dorsal side (1 point). Here these factors stabilize β-catenin by preventing its
degradation. The β-catenin/Wnt pathway is one of the signaling pathways required for organizer
formation (3 points).
The second signaling pathway necessary for formation of the organizer is TGF-β. The TGF-β family
members, Vg1, VegT, are vegetally localized (3 points).
The overlap of the two signaling pathways: TGF-β/Vg1/VegT and β-catenin/Wnt results in high
Nodal signaling in the dorsal most vegetal cells (endoderm). High Nodal signaling then induces the
overlying mesoderm to become the organizer (2 points). 1 point was given if the Nieuwkoop
Center in the dorsal endoderm was described to induce the over lying mesoderm to become
organizer.
(Description can be as above or include diagrams similar to below)
BMP antagonists such as chordin, noggin or follistatin are not acceptable answers. These
molecules are not involved in the formation of the organizer. Instead, they are secreted from the
organizer and are important for patterning the embryo along the dorsal-ventral axis.
E-55, Exam 2-2011: Name Page 8 of 9

F.
S.
39. (10 points). The notochord is primarily important for two processes in neural tube
development.
A. If you were to remove the notochord BEFORE neural tube folding, what cells/cell
types would fail to be induced?

The notochord is important for inducing the medial hinge cells (MHP) in the ectoderm (5 points).

1 point was deducted for only writing MHP.

Epidermis is not an accepted answer. This question refers to the removal of a notochord and not
the organizer. Therefore, the experiment is being performed on a later stage embryo (gastrulation
has completed), and the neural plate is already determined to become neural tissue. See Figure
L.
7.16, p255 about commitment of the neural tissue and p338 for the function of the notochord in
induction of the medial hinge point cells (MHP).

B. If you were to remove the notochord AFTER neural tube formation (folding has
completed), what cell/cell types would be missing?

The notochord is essential for inducing ventral neural tube fates such as the floorplate and motor
neurons (5 points).

2.5 points was given to people who said bone/sclerotome. Although we have not talked about this
in class, in our next lecture we will talk about how Shh from the notochord is important for
sclerotome induction.
E-55, Exam 2-2011: Name Page 9 of 9

______________________________________________________________________

F.
S.
L.