BIRLA INSTITUTE OF TECHNOLOGY AND SCIENCE, PILANI-

HYDERABAD CAMPUS
SECOND SEMESTER 2017-2018
BIO F341: Developmental Biology
Evaluation Component: Mid Semester (CB)
Date: 12.3.2019 Maximum Marks: 60 (weightage 30%) Duration: 90 Min

1. Imagine you have discovered a novel transcription factor Goosecoid; it is involved in

mesodermal fate specification in chicken. You now want to identify all the genes
regulated by it in the chicken genome. Name the technique you would use and explain its
principle. (2+ 4 M)
 
Name of the technique is Chromatin Immunoprecipitation- Sequencing (ChIP- Seq/ Chip-chip)
 
Chicken Embryo at a given embryonic stage are treated with formaldehyde so that proteins
bound to the cis-regulatory control of DNA are chemically linked.
 
Isolate and shear the DNA. Incubate this sheared chromatin with Ab specific for Goosecoid. This
will pull down or precipitate all the DNA fragments to which Goosecoid was bound in the
embryo.
 
The identity of DNA can be determined by digesting the protein away followed by sequencing or
DNA microarray.
 
The associated gene can then be identified by searching the database of chicken genome
sequence.

2. Convergent extension is the process by which sheet of cells changes shape by extending
in one direction and narrowing in direction at right angles to the extension, caused by the
cells intercalating between each other. Design an experiment to demonstrate convergent
extension during Xenopus Gastrulation. Also mention the expected result. (3+3 M)
Convergent extension occurs in mesoderm during Xenopus gastrulation. The mesoderm
converges along the midline to form the Notochord. Brachury is a mesodermal marker.
Immunostaining of the embryo with Brachyury Antibody at different stages of Gastrulation will
mark mesoderm.
At the start of Gastrulation Brachury expressing cells (mesoderm) will appear as narrow ring
around the blastopore. As gastrulation proceeds it will converge in to a narrow band along the
dorsal midline of the embryo extending in the A-P direction.
 Left: before gastrulation, the expression of Brachyury (dark stain) marks the future mesoderm, which is
present as an equatorial ring when viewed from the vegetal pole.

Right: as gastrulation proceeds, the mesoderm that will form the notochord (delimited by blue lines)
converges and extends along the midline

3. Match the following 12 M

A Gap gene i Gurken
B Pair-rule gene ii Acts in Wnt pathway
C Terminal system iii Primitive streak
D Maternal gene iv Kidney
E B-catenin v Nervous system
F Hensen’s node vi Kruppel
G Ectoderm vii Even skipped
H Mesoderm viii torso

Ans.
A Gap gene vi Kruppel
B Pair-rule gene vii Even skipped
C Terminal system viii torso
D Maternal gene i Gurken
E B-catenin ii Acts in Wnt pathway
F Hensen’s node iii Primitive streak
G Ectoderm v Nervous system
H Mesoderm iv Kidney
4. Define model organism. What characteristics of Drosophila make it model organism? (2
+ 4 M)
Model organisms have following characteristics:
Specific species or organism
Extensively studied in research laboratories
Advance our understanding of Cellular function, Development and Disease
Ability to apply new knowledge to other organisms
Characteristics of Drosophila that make it a good model organism
Small, easy and cheap to maintain and manipulate
Short lifespan
Produce large numbers of offspring
Development is external
Availability of mutants
Lots of history/previous experiments and discoveries
Genome is sequenced
Homologues for at least 75 % of human disease genes
Exhibit complex behaviours
Fewer ethical concerns

5. What is the first signaling center that develops in the dorsal-vegetal region in the
Xenopus blastula? How is it important in the determination of dorsal and anterior
structures? (4+6M)
First signaling center that develop in the dorsal- vegetal region in the Xenopus blastula is
known as the blastula organizer
or Nieuwkoop center.
Nieuwkoop Centre sets up D/V
polarity in the blastula.
The 1st cleavage cuts through
the site of sperm entry and the
Nieuwkoop Centre. The 2nd
cleavage splits the embryo into
4 cells (2 with the Nieuwkoop
Centre and 2 without).
To determine the dorsal and anterior structure, nieuwkoop centre grafts in 32 celled
embryo:
1) When cells of the Nieuwkoop Centre are grafted onto the ventral side, twinned
embryos with two 'dorsal sides’ are produced.
2) When cells of the ventral side are grafted onto the dorsal side, normal embryos are
produced (no effect). Therefore, signal from the Nieuwkoop Centre must be required for
developing dorsal and anterior structures.
6. Write the steps involved in construction of chimeric mice for the study of morphogenetic gene
with an example. (8M)

7. Segmentation genes determine the segment of the embryo in Drosophila and thus divide
the embryo into regions that correspond to the adult segmentation patterns. Name the
class of these genes and their specific role. (6 M)
Gap genes
i. Subdivide the embryo along the anterior-posterior axis.
ii. Mutation results in the deletion of several adjacent segments.
Pair rule genes
iii. Divide the the embryo into regions, each containing parasegments.
iv. Mutations cause deletions of the same part of a pattern in every other segment.
Segment polarity genes
v. Determine regions that become segments of larvae and adults
vi. Mutants possess parts of segments replaced by mirror images of adjacent half
segments.
8. Changes in the egg membrane at fertilization block polyspermy. Explain the mechanisms.
(6 M)
The resting membrane potential of the unfertilized sea urchin egg is -70 mV at
fertilization. It changes rapidly to + 20 mV, and then slowly returns to the original value.
This depolarization may provide a fast block to polyspermy.
The egg is surrounded by a vitelline membrane, which lies outside the plasma membrane.
Membrane bound cortical granules lie just beneath the egg plasma membrane. At
fertilization the cortical granules fuse with the plasma membrane and some of the
contents are extruded by exocytosis. These join with the vitelline membrane to form a
tough fertilization membrane, which then lifts off the egg surface and prevents further
sperm entry.