Slide 1

Slide 1 text

Peaks and valleys of prolactin-driven gene expression during parental care Dr. Rayna M. Harris raynamharris Postdoc, UC Davis 1

Slide 2

Slide 2 text

Meet the co-authors (*here at SICB) Dr. Rayna Harris Dr. Suzanne Austin Dr. Andrew Lang Dr. Matthew MacManes Dr. Rebecca Calisi Rechelle Viernes Dr. Jesse Krause April Booth Victoria Farrar University of California, Davis University of New Hampshire * * * Funded by NSF IOS-Animal Behavior. We are grateful to the members of our labs and and the labs of John Wingfield and Titus Brown lab for discussion. 2 *

Slide 3

Slide 3 text

Like humans, Rock Doves are bi-parental. However, both male and female Rock Doves lactate. Image by Victoria Farrar Calisi & a Working Group of Mothers in Science 2018 PNAS 3 Calisi 2018 Science

Slide 4

Slide 4 text

What molecular profiles characterize parental care transitions in the reproductive axis (HPG) of bi-parental Rock Doves? 4 https://github.com/macmanes-lab/DoveParentsRNAseq

Slide 5

Slide 5 text

Experimental design for characterizing HPG gene activity over the course of bi-parental care. 5 https://github.com/macmanes-lab/DoveParentsRNAseq N= 12 per group 576 RNA-seq samples

Slide 6

Slide 6 text

What explains the most variation in gene expression? Tissue, sex, or treatment? 6 https://github.com/macmanes-lab/DoveParentsRNAseq Kallisto -> Limma -> Rtsne or PCA

Slide 7

Slide 7 text

Gene expression in each tissue is quite distinct. 7 https://github.com/macmanes-lab/DoveParentsRNAseq Kallisto -> Limma -> Rtsne or PCA

Slide 8

Slide 8 text

Sex differences are most pronounced in the gonads. 8 https://github.com/macmanes-lab/DoveParentsRNAseq Kallisto -> Limma -> Rtsne or PCA

Slide 9

Slide 9 text

9 https://github.com/macmanes-lab/DoveParentsRNAseq Treatment effects are most pronounced in the pituitary. Kallisto -> Limma -> Rtsne or PCA

Slide 10

Slide 10 text

What gene explains the most variation? Prolactin. 10 https://github.com/macmanes-lab/DoveParentsRNAseq PC loadings calculated and visualized with `factoextra`

Slide 11

Slide 11 text

Confirms that prolactin is an important regulator of reproductive axis and parental care behavior. 11

Slide 12

Slide 12 text

Levels of prolactin, in the the blood are correlated with prolactin (PRL) expression in the pituitary. 12 https://github.com/macmanes-lab/DoveParentsRNAseq DESeq2: pituitary only, treatment * sex

Slide 13

Slide 13 text

13 Imagine PRL expression as a song. What genes work in concert to create a “transcriptional symphony”? https://github.com/macmanes-lab/DoveParentsRNAseq Made with `ggplot`. Played lived on a keyboard.

Slide 14

Slide 14 text

WGCNA identified 96 genes that are coexpressed with PRL in the pituitary. ShinyGO categorized these genes by their Biological Process. 14 https://github.com/macmanes-lab/DoveParentsRNAseq Thanks to Dr. Sarah Davies for the WGCNA suggestion via Twitter :)

Slide 15

Slide 15 text

The top GO terms are related to the cell cycle process, cell proliferation, reproduction, and stress response. 15 https://github.com/macmanes-lab/DoveParentsRNAseq Genes identified with WGCNA. GO terms identified with ShinyGo.

Slide 16

Slide 16 text

BRCA1 helps repair DNA. Mutations can lead to increased risk for breast cancer. BRCA1 peaks when PRL peaks around the time chicks hatch. 16 https://github.com/macmanes-lab/DoveParentsRNAseq

Slide 17

Slide 17 text

Understanding the dynamics of PRL and BRCA1 during avian parental care could advance cancer research. 17

Slide 18

Slide 18 text

RNA-seq is not just for data-driven discovery. It can be used to test hypotheses too. 18

Slide 19

Slide 19 text

Hypotheses Hypothesis 1 (H1) Changes in transcription during parental care are based on an internal clock-timing mechanism. 19 Hypothesis 2 (H2) Changes in transcription during parental care are based on external sensory information, like the the presence or absence of eggs or chicks.

Slide 20

Slide 20 text

Egg and chick replacement manipulations. 20 https://github.com/macmanes-lab/DoveParentsRNAseq

Slide 21

Slide 21 text

When chicks hatch early, gene expression is very similar to incubation to day 9 and very different from normal hatch. 21 https://github.com/macmanes-lab/DoveParentsRNAseq DESeq2: treatment * sex log fold change pituitary H1 H2

Slide 22

Slide 22 text

All three manipulations are more similar to their calendar day equivalent than to their external environment. 22 https://github.com/macmanes-lab/DoveParentsRNAseq DESeq2: treatment * sex log fold change pituitary H1 H2 H1 H2 H1 H2

Slide 23

Slide 23 text

Egg and chick removal manipulations. 23 https://github.com/macmanes-lab/DoveParentsRNAseq

Slide 24

Slide 24 text

Offspring removal manipulations also suggest that transcription is regulated by an internal clock. 24 https://github.com/macmanes-lab/DoveParentsRNAseq

Slide 25

Slide 25 text

And prolactin (PRL) still explains most of the variation. 25 https://github.com/macmanes-lab/DoveParentsRNAseq

Slide 26

Slide 26 text

This suggests that pituitary gene expression is driven by an internal clock that regulates prolactin-driven expression of parental care behaviors. 26

Slide 27

Slide 27 text

27 Thanks for listening. What questions do you have? Dr. Rayna M. Harris raynamharris Postdoc, UC Davis