Transcript study guide

New research on the hypothalamic-pituitary-gonadal axis:
H. Hasuwa et al., Science 341: 71-73, 2013
• Title: “miR200b and miR429 function in mouse ovulation
and are essential for female fertility”
• Review: hypothalamic-pituitary-gonadal axis
• Main question: how does GnRH promote LH secretion?
• Review: control of gene expression
What are microRNAs (miRNAs)?
Stefanie S. Jeffrey, Nature Biotechnology 2008
Genetic tools/tricks: reporter assay to see how
miR-200b & miR-429 affect Zeb1 expression
• What experiment was
done to yield the data
in the bar graph?
Red fluorescence
• What do the bars
show?
• Is it surprising that
Zeb1 expression was
knocked down the
most by miR-200b and
miR-429? Explain.
Hasuwa et al. (2013), Figure S13
Genetic tools/tricks: overexpressing Zeb
• What is Zeb1?
• Is the Fshb a strong
promoter or a weak
promoter? How can you
tell?
Google Images search results;
Hasuwa et al. (2013), Figure 3B-C
Literature-reading tip:
figure (and table) titles often collectively tell the story!
• Fig. 1. Infertility phenotype of miR-DKO mice.
• Fig. 2. Pituitary defects in miR-DKO female mice.
• Fig. 3. Lhb expression is regulated by ZEB1 as a
potential target of miR-200b and miR-429.
Fig. 1. Infertility phenotype of miR-DKO mice.
• What does the Y axis mean?
• What do the 3 right-most bars tell us?
Hasuwa et al. (2013), Figure 1A
Fig. 1. Infertility phenotype of miR-DKO mice.
• How was the experiment in panels C and D different from the
experiment in panels E and F?
• What were the results in each case?
Hasuwa et al. (2013), Figure 1C-F
Fig. 2. Pituitary defects in miR-DKO mice.
• What does DKO mean
here?
• This is a figure about
the pituitary. Why are
estradiol and
progesterone included?
Hasuwa et al. (2013), Figure 2A-D
Fig. 2. Pituitary defects in miR-DKO mice.
• Which is the anterior pituitary and which is the posterior pituitary? How can you
tell?
• How is Figure 2G-H similar to and different from Figure 2B?
Hasuwa et al. (2013), Figure 2E-H
Fig. 3. Lhb expression is regulated by ZEB1 as a
potential target of miR-200b and miR-429.
D: What was the experiment?
What was the result?
E: The red, purple, and black bars are all small
relative to the blue one. Is this surprising?
Explain.
Hasuwa et al. (2013), Figure 3D-E
Create a diagram to summarize the main
findings of this study. Include LH, Zeb1, and
the miRNAs.
What questions could be researched next?