committing the gonad to ovary fate - comtecgroup ppt... · 2014-11-20 · anirudh natarajan xx log...
TRANSCRIPT
Committing the Gonad to
Ovary Fate
Blanche Capel
Department of Cell Biology
Duke University Medical Center
The testis and ovary arise from the same primordia
E10.0-11.5
(mouse)
granulosa cells supporting cell lineage
steroidogenic cell lineage
primordial germ cell lineage spermatogonia
Leydig cells
Sertoli cells
Theca cells
oogonia
The bipotential gonad is composed of three bipotential lineages
Anne McLaren Paul Burgoyne Robin Lovell-Badge
The supporting cells are primed with an ovarian bias
Expression of the Y-linked Sry gene in the
supporting cell lineage drives testis fate
E10.0-11.5
(mouse)
E12.5
testis ovary
bipotential
Sry
The supporting cells are the first to show sexually dimorphic expression
a small number of transcripts are dimorphic in germ cells and all are X- or Y-linked
many genes have a pattern specific to supporting cell precursors that is shared between XX and XY cells
Sample global gene expression in gonads at 6 time points between E11.0-E12.0
Steve Munger
Sample gonads every 4 hours
Data from the time course revealed patterns of activation and repression in XX and XY gonads
XX Log Fold Change
XY Log Fold Change
Sox9
Dhh
Cbln1
XY Log Fold Change
Most male genes are activated in XY gonads
Anirudh Natarajan
XX Log Fold Change
Msx1
Wnt4
Irx3
Many female genes are repressed in XY gonads
XX Log Fold Change
XY Log Fold Change
Cascades of Dimorphic Expression –
Male-enriched genes
Fgf9
Acer3
Amh
Expression Trajectory
Cascades of Dimorphic Expression –
Female-enriched genes
Fst
Zfp277
Igfbp2
Expression Trajectory
XX XY
bipotential
?
But do adult
granulosa cells
directly stem
from this
population?
Adult Sertoli
cells develop
from supporting
cell precursors
Some female somatic cells can activate the Sry
promoter
GATA4
Sry-EGFP
Sry promoter
EGFP
The same cells also express FOXL2, a granulosa cell
marker
E13.5
Mork et al. Biol Reprod 2011
Sry-EGFP
FOXL2
Sertoli cells arise from the coelomic epithelium
during the bipotential period
After Karl et al., 1998
+ 2 h
+ 6 h
E11.75 +24 h E11.5 + 24 h
Mitotracker SOX9
Granulosa precursor cells arise from the XX
coelomic epithelium until E14.5
Labeled at E11.5-13.5
Mork et al. Biol Reprod 2011
trace 2
trace 3
birth
trace 1
trace 3
FOXL2+ cells are
mitotically arrested
from the time they
leave the surface
epithelium
E12.5
Mork et al. Biol Reprod 2011
Cyclins are specifically depleted in XX relative to XY supporting cells
FOXL2+ cells do not resume cycling until their
host follicle is activated
P1 P7
Ki67 FOXL2
p27 FOXL2
Follicle assembly
Do fetal FOXL2 cells form adult granulosa cells?
trace2
birth
sex determination
trace 1
Foxl2-
GFPCreER
R26R-lacZ
inject tamoxifen
Foxl2-GFPCreER; R26R-lacZ
Mork et al. Biol Reprod 2011
Fetal Foxl2-expressing-cells contribute granulosa cells to
medullary but very rarely cortical follicles
Two types of granulosa cells for two types of
follicles?
Cortical granulosa
cells
Medullary granulosa
cells
Cortical follicles are activated
during adult estrus cycles
Medullary follicles are
activated immediately after
follicle assembly at birth;
establish ovarian functionality
at the hormonal level prior to
the first LH surge
Follicle assembly
trace 2
trace 3
birth sex determination
Where do the cortical granulosa cells come from?
trace 1
BrdU Trace
Cortical granulosa cells arise from cycling
progenitors
Mork et al. Biol Reprod 2011
The ovarian surface epithelium gives rise to
pregranulosa cells between P1 and P7
Mork et al. Biol Reprod 2011
+ 24 h +72 h +96 h +96 h
P1
P3
P7
p27 DNA
Once specified, pregranulosa cells immediately
enter cell cycle arrest
Summary of mouse granulosa cell origins
Mork et al. Biol Reprod 2011
Ng et al., Nature Cell Biology, 2014 Rastetter et al, DevBio, 2014
Lgr5+ cells are present on the ovarian surface from E12.5 to adult stages, and act as progenitors for granulosa cells
Danielle Maatouk
• TSS/promoters • enhancers/repressors
• insulators
• LCRs
Isolate Supporting Cells for DNase Hypersensitivity Study
Sertoli-specific sites are identified by comparison to 6 other datasets
Enhancers of genes expressed in granulosa cells also have Sertoli-specific sites
Perhaps shared epigentic genome marks underlie
transdifferentiation of Sertoli cells into
granulosa-like cells in adult testes of Dmrt1 mutants
Matson et al. 2011
Transdifferentiation of granulosa cells into
Sertoli-like cells in adult ovaries
Uhlenhaut et al. 2009
Couse et al. 2003
SOX9
Adult induced Foxl2 KO
ERαβ KO
SOX9
THANKS Collaborators
Uwe Uhler – dcsb, Duke Greg Crawford – Duke
Steve Munger Samantha Jameson Anirudh Natarajan Danielle Maatouk Lindsey Mork Iordan Batchvarov Help from many others in the lab
NICHD, NIDDK
Duke Center for Systems Biology