Matthew Dean

Early Embryonic Loss(55%)

(Zinaman et al. 1996; Diskin et al. 2006; Wilcox et al. 1988)

Fertilization Failure(5%)

Live Birth(33%)

Late Embryonic/Fetal Loss

(7%)

Early Embryonic Loss(55%)

Fertilization Failure(5%)

Live Birth(33%)

Late Embryonic/Fetal Loss

(7%)

(Zinaman et al. 1996; Diskin et al. 2006; Wilcox et al. 1988)

Early Embryonic Loss(55%)

Fertilization Failure(5%)

Live Birth(33%)

Late Embryonic/Fetal Loss

(7%)

(Zinaman et al. 1996; Diskin et al. 2006; Wilcox et al. 1988)

Early Embryonic Loss(55%)

Fertilization Failure(5%)

Live Birth(33%)

Late Embryonic/Fetal Loss

(7%)

(Zinaman et al. 1996; Diskin et al. 2006; Wilcox et al. 1988)

Early Embryonic Loss(55%)

Fertilization Failure(5%)

Live Birth(33%)

Late Embryonic/Fetal Loss

(7%)

(Zinaman et al. 1996; Diskin et al. 2006; Wilcox et al. 1988)

Zygote

Uterus

Cervix

Vagina

Oviduct

(Hansson 1947; Senger 2005)

Ovary

Morula

Uterus

Cervix

Vagina

Ovary

Oviduct

(Hansson 1947; Senger 2005)

BlastocystUterus

Cervix

Vagina

Ovary

Oviduct

(Hansson 1947; Senger 2005)

Oviduct

Uterus

Cervix

Vagina

Histotroph:•Hormones•Nutrients•Glucose

(Bazer et al. 2012)

Oviduct

Uterus

Cervix

Vagina

Histotroph:•Hormones•Nutrients•Glucose

(Bazer et al. 2012)

EndometriumUterine Lumen

(Bazer et al. 2012; Yánez et al. 2003)

Glucose

Glycogen

Pyruvate

X

Glycogen

Glycogen

glucose

-6PHexokinase

Glycogen

glucose

-6PHexokinase

Glycogen

glucoseCellular Metabolism

-6P -UDPHexokinase

Glycogen

glucoseCellular Metabolism

-6P -UDPHexokinase

Glycogen Synthase

Glycogen

glucoseCellular Metabolism

-6P -UDP -1PHexokinase

Glycogen Synthase

Glycogen Phosphorylase

Glycogen

glucoseCellular Metabolism

-6P -UDP -1PHexokinase

Glycogen Synthase

Glycogen Phosphorylase

-6P

Glycogen

glucoseCellular Metabolism

-6P -UDP -1PHexokinase

Glycogen Synthase

Glycogen Phosphorylase

-6P

Glucose-6-Phosphatase

Glycogen

glucoseCellular Metabolism

-6P -UDP -1PHexokinase

Glycogen Synthase

Glycogen Phosphorylase

-6P

Glucose-6-Phosphatase

Glycogen

glucoseCellular Metabolism

Estrus/

Mated

Glyc

og

en

C

on

ce

ntratio

n

(m

g/ 1

00

g

w

et tissu

e)

(modified from Demers et al. 1972: Gupta et al. 2013)

Day Post Mating

Implantation

Estrus

0 1 2 3 4

Estrus

1 2 3 4 5

1 2 3 4 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 … 50

ImplantationEstrus

Implantation

Day

Day

1) Characterize changes in uterine glycogen metabolism during estrus, embryonic diapause, and pregnancy in mink.

2) Determine if the glycogenic effect of estradiol on uterine epithelia is direct.

Feb AprilMarch May

15 20 25 5 10 15 20 25 30 5 10 15 20 25 30 5 10 15

Estrus Implantation Parturition

EstrusDiapause

Pregnancy

3 Collections

Objective 1.Characterize changes in uterine glycogen metabolism during estrus,

embryonic diapause, and pregnancy in mink.

0

5

10

15

20

25

30

(Nested ANOVA A-CP<0.05)

Estrus IIS IS

A

B

C

Gly

cog

en

Co

nce

ntr

ati

on

(mg

/mg

tis

sue

)

DiapausePregnancy

Myometrium

Endometrium

Periodic Acid-Schiff-stain for carbohydrates

0

10

20

30

40 A

B

C

(ANOVA and Tukey’s, P<0.05)

Pe

rce

nt

Are

a

Estrus Diapause Pregnancy

Estrus

Diapause

Pregnancy

Myometrium

Endometrium

LE

GE

S

0

10

20

30

40

50

Glandular Epithelia

Lumenal Epithelia

Stroma

A

B B

[[

*

(Nested ANOVA A-B,* P<0.05)

Gly

coge

n C

on

ten

t(P

erce

nt

Are

a)

Estrus Diapause Pregnancy

Estrus

Diapause

Pregnancy

*

Estrus Diapause Pregnancy

Enzyme= BlueCounterstain=red/pink

-6P

Estrus Diapause Pregnancy

Gly

coge

n

Syn

thas

eG

lyco

gen

P

ho

sph

ory

lase

Enzyme= BlueCounterstain=red/pink

0

1

2

3

4

5

6

Re

lative

m

RN

A E

xp

re

ssio

n

C

B

A

Estrus Diapause Pregnancy

-6P

Estradiol???

Cont Estradiol

Perc

ent

Are

a

0.0

0.1

0.2Glandular Epithelial Glycogen

*

Estrus

EstradiolEstrus

EstradiolEstrus

?

1) Characterize changes in uterine glycogen metabolism during estrus, embryonic diapause, and pregnancy in mink.

2) Determine if the glycogenic effect of estradiol on uterine epithelia is direct.

Mink uterine epithelia

Immortalized with SV40 large T antigen

Positive for cytokeratin 18

(Moreau et al. 1995; http://micro.magnet.fsu.edu)

Cytokeratin 18uteru

sG

MM

e

Mucin 1

uteru

sG

MM

e

HexokinaseGlycogen Synthaseu

terus

GM

Me

uteru

sG

MM

e

Glycogen Phosphorylase u

terus

GM

Me

uteru

sG

MM

eGlucose-6-

Phosphatase

0 0.1 1 10 100

0.0

0.1

0.2

0.3

Gly

co

gen

(g

/10

6 c

ells)

Estradiol (nM)

Insulin (g ml-1)0 1.8 5.5 16.6 50

Gly

co

gen

(

g/1

06 c

ells)

0.0

0.2

0.4

0.6

**

(*P<0.05 relative to 0 μg ml-1)

PPPT- Igf1r antagonist(*P<0.05 relative to 0 μg ml-1)

Gly

co

gen

(

g/1

06 c

ells)

0.0

0.1

0.2

0.3

Cont PPPT Insulin Insulin+PPPT

*

InsulinEstradiol

00

500

500.1

501.0

5010

50100

Gly

co

gen

(

g/1

06 c

ells)

0.0

0.2

0.4

*

*

(*P<0.05 relative to 0 μg ml-1 insulin & 0 nM estradiol)

ICI= Estrogen Receptor antagonist(*P<0.05 relative to cont

Glycogen

CG

lyc

og

en

(

g/1

06 c

ells

)

0.0

0.2

0.4

0.6 *

EstradiolICICont Estradiol+ ICI

1)Endometrial glycogen contributes to histotroph.

localized to epithelial at estrus

mobilized before implantation

high glucose-6-phosphatase during diapause

2) Estradiol stimulates glycogenesis through both paracrine and direct mechanisms.

activation of IGF-1 receptor increases glycogen

estradiol increases glycogen (in presence of insulin)

Estradiol

IGF1

At Estrus

GlycogenStroma Epithelia

Estradiol

IGF1

At Estrus

GlycogenStroma Epithelia

Estradiol

IGF1

Pre-Implantation

GlycogenStroma Epithelia

Estradiol

IGF1

Pre-Implantation

GlycogenStroma Epithelia

Progesterone4-hydroxyestradiol

CommitteeDr. Rose

Dr. Bearden

Dr. Cretekos

Dr. Pfau

Lab MatesJason Hunt

Kole Bowman

Lisa McDougall

Josh Astle

Ayokunle Hodonu

Evan Stegelmeier

Other Grad Students

Other ProfessorsDr. Rodnick

Dr. Aho

Dr. Thomas

Funding SourcesMoyle Mink and Tannery

Fur Commission USA

Idaho INBRE

ISU Research Council

MCRF Seed Grant

Sigma Xi

Thank you