Genevieve Neal-Perry, MD PhD
Associate Professor Ob/Gyn and Neuroscience
Fellowship Director, Division of Reproductive Endocrinology and Infertility
Albert Einstein College of Medicine and Montefiore Medical Center
1)
2)
To describe vitamin D3 physiology
To discuss the differential impact of in
utero + perinatal vs. pubertal vitamin D3
deficiency on
a. puberty
b. reproductive physiology
i. Hypothalamus
ii. Pituitary
iii. Ovary
1)
2)
Fat soluble steroid
hormone
 vitamin D3 steroid
hormone receptor
(VDR)
 RXR
Genomic vs. nongenomic
non-classical
classical
UV-B 20-30
min 2x wk
Calicidiol
½ life =15 d
Caltriol
½ life =15 h
Group
FDA
Safe
Toxic
Recommend High
Dose
Dose (IU)
FDA (IU)
Infants
0-12
months
400*
<1000
1000-1500 400-800
Children
600*
2000
≥30004000
Pregnant
Women
600*
2000
≥4000
Lactating
Women
600*
10,000
per
day**
Adults <50 600*
Adults
51-70
Best Dose
800-1000 D3 /day
50,000 D2/2 week
50,000 D2/month
600*
Adults >70 800*
*RDA and IOM recommendations
**safe for at least 5 months
Internation Units (IU)
Dietary Sources of Vitamin D3
Food
2000
Cod Liver Oil
Cooked
1500 Salmon
Cooked Mackerel
Canned Tuna Fish
1000
WarningVitamin
Label !
Serving
D
To achieve the daily recommendation of
600 IU of vitamin D you have
to eat:
(IU)
3
30 eggs = 50 cubes of Swiss cheese
Tbsp
1360
=1201oz
of cereal = 6 cups
milk
3 ½ oz
3 ½ oz
3 oz
Canned Sardines
1 ¾ oz
500 OJ
Fortified
8 oz
Fortified Milk
8 oz
0 Sunshine
Fortified
cerealCod liver oil Cooked
8 oz Milk
(tbsp)
(8 oz)
salmon
Egg (yolk)
1
egg
(3 ½ oz)
360
345
200
Cereal
(8 oz)
250
100
98
40
Egg
Yolk
20
Swiss
cheese
( 1 oz)
Health Implications
Status
Deficient
Insufficient
Adequate
Toxic
Serum 25(OH)D3
nmol/liter
ng/ml
<30
<12
30-49
16-19
50-124
20-49
>125
>50
1)
People with dark skin pigmentation
 African American > Hispanic > Caucasian
2)
3)
4)
Obese individuals
Elderly
Reproductive-aged women, neonates,
and adolescents
% <50 nmol/L vit D3
Yetley, Am J Clin Nutr 2008:88(2): 558S National
Health and Nutrition Examination Survey 2000-2004
Yetley, E. A Am
J Clin Nutr 2008;88:558S
“2 out of every 3 pregnant women in
the U.S. have suboptimal vitamin D
status, with even higher prevalences
reported among African-American and
Hispanic women”
Increased medical
costs!!
[Looker, et al. Am J Clin Nutrition 2008]
PCOS and
Obesity
Small for
gestational
age
Vitamin D3
deficiency
Infertility
Recurrent
pregnancy
loss
Overarching Objective
To determine the mechanism (s)
by which VD3 regulates female
reproduction
Overarching Hypothesis
Developmental VD3 deficiency
disrupts hypothalamic-pituitarygonadal physiology in females
VDR and α-hydroxylase (cyp27b1) is widely
distributed in reproductive tissue
Vitamin D3 is necessary for neuronal
development
Vitamin D3 deficiency in humans is associated
with reproductive dysfunction
Rodent models of vitamin D3 deficiency
1)
2)
3)
4)
a.
b.
diet induced vitamin D3 deficiency in rats
Transgenic mice
1)
2)
Transgenic
cyp27b1 null
mice
Wild type
littermates
Group 1: Control WT and KO (VD3 sufficient diet throughout)
Group 2: KO in utero, perinatal and peripubertal VD3 deficient diet
Group 3: KO in utero and perinatal VD3 deficient diet
Group 4: KO peripubertal VD3 deficient diet
Group 2
Group 3
preconception intrauterine
conception
Group 4
perinatal
birth
peripubertal
weaning
on day 21
1) Vaginal opening
2) 1st estrus
3) Estrous cycling
Mice exposed to in utero, perinatal
and peripubertal VD3 deficiency
have a delayed puberty
1st estrus
Group 2
VO
preconception
in utero
conception
Days between VO
and 1st estrus
perinatal peripubertal
birth
Weaning
D21
VO
1st Estrus
Will in utero and perinatal VD3
deficiency disrupt puberty?
Group
3
1st estrus
VO
preconception
in utero
conception
Days between VO
and 1st estrus
perinatal
birth
peripubertal
weaning
D21
VO
1st Estrus
Will peripubertal VD3 deficiency
disrupt puberty?
Group 4
VO
preconception
1st estrus
In utero
perinatal
conception
birth
Days between VO
and 1st estrus
peripubertal
Weaning
D21
VO
1st estrus
Genetic
background
Modified from Roa et al.,
Molecular and Cellular Endocrinology 324(2010):87
BMI (g/mm2)
0.06
0.04
Postnatal growth
WT (D+)
KO (D+)
KO (D-)
0.02
0.00
0
1
2
3
Postnatal age (wk)
4
Average cycle length
(days)
15
Estrous Cycle Length
*
10
5
0
WT
KO(D-)
KO(D+)
P
(ng/ml)
Prl
(ng/ml)
E2
(pg/ml)
LH
(ng/ml)
FSH
(ng/ml)
* p<0.05
Estrus
Diestrus-1
Diestrus-2
Proestrus
Estrus
LH surge
LH surge
WT VD3+
P
E
D1
D2
10
Day
15
20
estrous stage
P
estrous stage
estrous stage
KO VD 3+
KO VD3-
P
5
LH surge
E
D1
D2
5
10
Day
15
20
E
D1
D2
5
10
Day
15
20
40
30
20
10
0
40
WT
(D)
Proestrus
20
a
10
(E)
WT
Diestrus II
40
30
a
20
10
0
WT
KO (D-) KO (D+)
Estrus
40
30
a
20
10
0
KO (D-) KO (D+)
KO (peripubertal Vit D 3-)
(F)
WT
KO (D-) KO (D+)
KO (rescue Vit D 3+)
P
estrous stage
50
KO (D-) KO (D+)
30
0
% of time spent in
Diestrus II per 5 days
(B)
% of time spent in
estrus per 5 days
% of time spent in
diestrus I per 5 days
Diestrus I
50
D2
estrous stage
(C)
% of time spent in
proestrus per 5 days
(A)
E
D1
D2
0
5
10
Day
15
D1
E
P
0
5
10
15
Day
a, P<0.01 vs. WT; b, P<0.05 vs. D+
Female reproductive physiology
Communication between the brain
and the ovaries change with
increasing reproductive age.
estradiol
ovarian egg
development
Progesterone
VD 3 deficiency does not affect ovarian
responsiveness to superovulation
KO (D-) + saline
KO (D-) + PMSG + hCG
CL
CL
CL
CL
CL
Follicle Stimulating
Hormone
N= 3-8
Negative Control
GT1-7 Kidney
GT1-7
Kidney
VDR
Peripubertal VD3 deficiency does not
affect GnRH neuron density
VD3+
GnRH ir cells
80
VD3 deficiency does not affect
the number of GnRH neurons
60
40
20
0
VD3-
in utero
lactation
peripubertal
D+
D+
D+
D+
D+
D-
1)
Delays the puberty
2)
Reversibly disrupts estrous cyclicity
3)
Does not primarily affect ovarian
physiology
4)
Does not affect GnRH neuron density
5)
Does not disrupt negative feedback
effects of estradiol during diestrus
6)
GT1-7 neurons express VDR
1)
2)
Changes in dietary habits and reduced
sun exposure have resulted in near
epidemic levels of VD3 insufficiency &
deficiency
Vitamin D3 insufficiency & deficiency
disproportionately affect reproductive
aged women, neonates and adolescents
thereby making these groups more
susceptible to the adverse consequences
of VD3 deficiency on the reproductive axis
Acknowledgments
Cary Dicken, MD
Joe Davis, MD
Yan Sun, MD PhD
Jun Shu, MD
Davelene Israel PhD
Anne Etgen, PhD
Nanette Santoro, MD
John Hardin MD
Streamson Chua, MD PhD
Cary Dicken
REI fellow
Jun Shu,
technician
Support
Yan Sun
R21HD0663511
Post Doc
Zondek Award for Reproductive Research
Department of Ob/Gyn Einstein College of Medicine
Joe Davis
REI fellow