How to read a research article:

1. Read the abstract to get an idea of what the paper is about and what are the major findings.

2. Read the introduction to understand the background and identify the question/goal/hypothesis
the paper addresses/makes.

3. Look at the figures. Usually one figure will refer to one question/concept the researchers are
addressing. A good paper is written so you can understand most of it by understanding the figures.

4. The results section will give you more detail on each figure. For each figure identify the
following:
a. The question the researchers are asking.
b. How are they doing the experiment?
c. What is/are the result/s?
d. What is/are the conclusion/s?
e. Do the results fully support the conclusions?
f. Are there any other experiments the researchers would need to do to support their
conclusions?

5. After reading/understanding all the figures decide if this work answered the overall
question/goal/hypothesis.

6. Look at the discussion to get a feeling of how this work fits in the big picture and to identify
future directions. You should be thinking of future directions also on your own and not just what
the paper suggests.

7. Read the Methods if you want to know exactly how the experiment was done or if you need to
use similar methods in your own research.
PDBIO 382 Name______Joseph Bohman_________________
Worksheet 1

You are encouraged to work in groups to discuss the answers to the following questions. However,
please write the answers in your own words. You will not be given credit if you copy the answers
from someone else.

1. What are placodes? (0.5 points)

It is a precursor of ectoderm that eventually differentiates into important peripheral nervous system
elements.

2. What is the significance of using Pax3 in this study? In other words, what is Pax3? (0.5 points)

A molecular marker for the trigeminal placode. It is a transcription factor and there is a Pax-3 protein
expressed as a result of the transcription factor.

3. What is the purpose/goal of this study? (0.5 points)

Which regions respond to Pax-3 transcription factor to express Pax-3 protein, when is the placode
ectoderm is specified, and where Pax-3 is present.

4. When is a tissue competent? What experiments in this article illustrate the concept of competence?
Summarize (Briefly!) the results and conclusions. Do the results fully support the conclusions, or are
the conclusions overstated? (1.5 points)

A tissue is competent when it responds to the Pax 3 TF, and expresses the Pax-3 protein in order to
develop the trigeminal placode. They grafted quail ectoderm into chick host at midbrain to see which
ectoderm was competent to respond and express Pax-3. They also did time-dependent experiments
marking the levels of Pax-3 in different stages of development to determine when tissue is committed to
expressing the protein. They found that before 3-ss Pax-3 induction signaling starts, and the head
ectoderm rostral to first somite is competent to respond to inducing, but going down past the otic region it
does not respond. The neural tube induces pax-3 expression.
5. When is a tissue specified? What experiments in this article illustrate the concept of specification?
Summarize (Briefly!) the results and conclusions. Do the results fully support the conclusions, or are
the conclusions overstated? (1.5points)

A tissue is specified when it does not require outside signals from other tissue in order to cause it to
express certain protein. They explanted or took tissue from developing quail in different stages of
development, starting with 3-ss, and put them separate from signal inducing tissues nearby. They found
that the cells started to express Pax-3 a lot by 8-9-ss, so they had already been specified by 3ss. The
conclusion states this regarding stages of development, where they say that specification has happened
before 3ss.

6. What is induction? What experiments in this article illustrate the concept of induction? Summarize
(Briefly!) the results and conclusions. Do the results fully support the conclusions, or are the
conclusions overstated? (1.5 points)

When one tissue signals another tissue to have a distinct cell fate. In Figures 5 and 6, this is shown when
they take midbrain-level ectoderm from quail and transplant it all the way down the neural tube in a chick
embryo. They then measured levels of Pax-3 expression and found that all of the midbrain-level ectoderm
from the quail donor expressed Pax-3, meaning the competent tissue was induced no matter where it was
on the neural tube. This supports their conclusion that the neural tube and the entire neural axis can induce
Pax-3 expression.

7. When is a tissue committed? What experiments in this article illustrate the concept of commitment?
Summarize (Briefly!) the results and conclusions. Do the results fully support the conclusions, or are
the conclusions overstated? (1.5 points)

When the differentiation is irreversible, then it is committed. This is different from specified tissues,
because if specified tissue receives other signals it can change its cell fate. Fig 4 shows the results of an
experiment they conducted where they explanted quail tissue in different stages of development into chick
tissue where the induction of Pax-3 is thought to be absent. They observed that past 8ss the cells started to
express a lot of Pax-3, and determined that after 8ss the cells can be thought to be fully committed.

8. Figures 1, 2, and 3 depict quail-chick grafting experiments. Name and define the developmental
process or state being tested for in each of these figures, according the article. (1 point)
 Figure 1 Figure 2 Figure 3

Figure 1: Specificity
Figure 2: Commitment
Figure 3: Induction

9. What could be the “next step” in placode cell fate research? What experiments would you do?
(1.5 points)

Determine other needed molecular markers for placode development. Knockout Pax-3 using Crispr
and see if development of the trigeminal placode still proceeds, and observe any differences. Using Crispr
add Pax-3 TF to other non competent cells and see if they will differentiate into trigeminal placode.