1. New research on the hypothalamic-pituitary-gonadal axis: H
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

2. What are microRNAs (miRNAs)?
Stefanie S. Jeffrey, Nature Biotechnology 2008

3. 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?
What do the bars show?
Is it surprising that Zeb1 expression was knocked down the most by miR-200b and miR-429? Explain.
Red fluorescence
Hasuwa et al. (2013), Figure S13

4. 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

5. 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.

6. 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

7. 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

8. 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

9. 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

10. Hasuwa et al. (2013), Figure 3D-E
Fig. 3. Lhb expression is regulated by ZEB1 as a potential target of miR-200b and miR-429.
E: The red, purple, and black bars are all small relative to the blue one. Is this surprising? Explain.
D: What was the experiment? What was the result?
Hasuwa et al. (2013), Figure 3D-E

11. Create a diagram to summarize the main findings of this study
Create a diagram to summarize the main findings of this study. Include LH, Zeb1, and the miRNAs.

12. What questions could be researched next?