First published online 13 December 2006
doi: 10.1242/dev.02743
Development 134, 407-415 (2007)
Published by The Company of Biologists 2007
A uterine decidual cell cytokine ensures pregnancy-dependent adaptations to a physiological stressor
S. M. Khorshed Alam1,
Toshihiro Konno1,
Gouli Dai2,
Lu Lu1,
Danhua Wang1,
Judy H. Dunmore3,
Alan R. Godwin3 and
Michael J. Soares1,3,4,*
1 Departments of Pathology and Laboratory Medicine, Institute of Maternal-Fetal
Biology, Division of Cancer and Developmental Biology, University of Kansas
Medical Center, Kansas City, KS 66160, USA.
2 Departments of Pharmacology, Toxicology, and Therapeutics, Institute of
Maternal-Fetal Biology, Division of Cancer and Developmental Biology,
University of Kansas Medical Center, Kansas City, KS 66160, USA.
3 Departments of Molecular and Integrative Physiology, Institute of
Maternal-Fetal Biology, Division of Cancer and Developmental Biology,
University of Kansas Medical Center, Kansas City, KS 66160, USA.
4 Departments of Obstetrics and Gynecology, Institute of Maternal-Fetal Biology,
Division of Cancer and Developmental Biology, University of Kansas Medical
Center, Kansas City, KS 66160, USA.
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Fig. 1. Mouse Dprp gene, construction of a Dprp-null mutant
targeting vector, genotype analysis, and Dprp mRNA and protein
expression. (A) Exons 2-6 of the mouse Dprp gene were
replaced with an in-frame EGFP gene followed by an MC1neo
cassette. (B) PCR analysis of wild-type (+/+), heterozygous (+/-) and
null (-/-) alleles. (C) RT-PCR analysis of Dprp transcripts in
gestation day 7.5 decidua from wild-type (+/+) and Dprp-null (-/-)
mice. (D) Western blot analysis of DPRP protein in gestation day 7.5
decidua from wild-type (+/+) and Dprp-null (-/-) mice.
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Fig. 2. Dprp and DprpGFP allele expression in the
uteroplacental compartment in implantation sites of wild-type, heterozygous
and Dprp-null mutant mice. Immunostaining for DPRP (A-C)
and GFP (D-F) was performed on frozen sections from gestation day 7.5
implantation sites of wild-type (A,D), heterozygous (B,E) and homozygous
mutant (C,F) mice. (G-I) EGFP fluorescence is shown in sections from
gestation day 7.5 implantation sites of wild-type (G), heterozygous mutant (H)
and homozygous mutant (I) mice (counterstain, Propidium Iodide). The
mesometrial region of the uterus is located at the top of each image. Scale
bars: 1 mm.
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Fig. 3. Histological examination of mesometrial and anti-mesometrial decidua of
gestation day 11.5 wild-type and Dprp-null mice. Mesometrial
(A-C) and anti-mesometrial (D-F) decidua were monitored for DPRP
expression in wild-type (+/+) tissues (A,D), and for GFP in DPRP-null (-/-)
tissues using anti-GFP (B,E) and fluorescence (C,F). The arrowheads in A-C
indicate the location of the mesometrial decidua. Note the minimal GFP
expression in the mesometrial decidua of B and C. By contrast, the
anti-mesometrial decidual regions appear comparable in wild-type and
Dprp-null tissues. Scale bars: 250 µm.
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Fig. 4. Expression analysis of wild-type and Dprp-null mice.
(A) Northern analysis for Dprp, Plp-j, Plp-b and Mt1
in decidual tissues. Total RNA was isolated from decidual tissues of wild-type
(+/+) and Dprp-null (-/-) mice on day 7.5 of gestation.
Gapdh was used to demonstrate integrity of the RNA and loading
accuracy. (B) PRL superfamily expression patterns were examined in
mouse placentas using the PRL superfamily miniarray assay. cDNAs for all
members of the mouse PRL superfamily were spotted on to nylon membranes. Total
RNA from day 12.5 or day 17.5 placental tissues were used to make probes by
reverse-transcription. Gapdh and salmon sperm DNA were used as
controls. (C-F) Localization of Dprp (C,D) and Plp-j
(E,F) mRNAs in implantation sites of wild-type (+/+; C,E) and
Dprp-null (-/-; D,F) mice on day 7.5 of gestation. Dprp and
Plp-j plasmids were used as templates for the synthesis of
digoxigenin-labeled sense and anti-sense RNA probes. The sense probes did not
demonstrate specific staining (data not shown). The mesometrial region of the
uterus is located at the top of each image. Scale bars: 1 mm.
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Fig. 5. Decidualization responses in wild-type and Dprp-null mice.
(A,B) Gross appearance of artificially decidualized uteri from
day 7.5 pseudopregnant wild-type (+/+) and Dprp-null (-/-) mice.
(C) Day 7.5 pseudopregnant deciduoma weight responses from wild-type
(+/+; n=7) and Dprp-null (-/-; n=9) mice.
(D) Day 7.5 pseudopregnant deciduoma weight responses from wild-type
(+/+; n=7) and Dprp-null (-/-; n=9) mice expressed
by ratio to body weight. *, P<0.01. (E) Alkaline
phosphatase (AP) activities of day 7.5 pseudopregnant deciduoma from wild-type
(+/+; n=7) and Dprp-null (-/-; n=7) mice.
(F) Immunocytochemical localization of DPRP in the day 7.5
pseudopregnant-decidualized uterus from wild-type (+/+) mice. (G)
Immunocytochemical localization of GFP in the day 7.5
pseudopregnant-decidualized uterus from Dprp-null (-/-) mice.
(H) GFP fluorescence in the day 7.5 pseudopregnant-decidualized uterus
from Dprp-null (-/-) mice. The mesometrial region of the uterus is
located at the top of each image. (I) Northern blot analysis of
Dprp, Plp-j, Plp-b, Mt1 and Gapdh expression in deciduoma
from day 7.5 pseudopregnant wild-type (+/+) and Dprp-null (-/-) mice.
Scale bars: 1 mm.
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Fig. 6. Pregnancies in Dprp-null mice are vulnerable to maternal
hypoxia. (A) Determination of the ontogeny of decidual PRL family
expression by RT-PCR analysis. (B) Exposure of pregnant females to
hypobaric hypoxia (equivalent of 11% oxygen) from days 5.5 to 11.5 of
gestation. After day 11.5, the animals were returned to ambient conditions and
examined on day 17.5 of gestation. (C,D) Gross appearance of a
representative uterus from pregnant wild-type (+/+) and Dprp-null
(-/-) mice exposed to hypoxia. (E) Quantification of pregnancy outcomes
in wild-type (+/+; n=19) and Dprp-null mutant (-/-;
n=10) mice exposed to hypoxia. Numbers of healthy and dying/resorbed
conceptuses are significantly different between wild-type and
Dprp-null mutant pregnancies; P<0.01. Note that unlike
wild-type pregnant female mice, Dprp-null pregnant female mice do not
adapt effectively to hypoxia.
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Fig. 7. Gross inspection of decidua-placental compartments from wild-type and
Dprp-null pregnant mice exposed to hypobaric hypoxia. Wild-type
(+/+) and Dprp-null (-/-) pregnant mice were exposed to the
equivalent of 11% oxygen from days 5.5 to 11.5 of gestation. Mice were
sacrificed on day 11.5 of gestation and uteroplacental compartments were
dissected. The locations of hemorrhagic regions are encircled (yellow broken
line) within the dissected uteroplacental compartments. Note the presence of
prominent hemorrhagic areas in the Dprp-null (-/-) tissues.
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Fig. 8. Histological examination of day 11.5 uteroplacental compartments of
wild-type and Dprp-null mice exposed to hypobaric hypoxia.
(A-C) Wild-type (+/+) and (D-F) Dprp-null (-/-) mice
were exposed to hypobaric hypoxia. Tissue sections were stained with
Hematoxylin and Eosin (A,B,D,E) or by isolectin B4 histochemistry
(C,F). Note the enlarged mesometrial blood spaces (A versus D, arrowheads),
the overgrowth of trophoblast giant cells (B versus E, arrowheads), and the
compressed mesometrial decidua and enlarged chorioallantoic placenta (C versus
F, dashed black lines demarcate the thickness of the mesometrial decidual
layer) in the Dprp-null tissues. The mesometrial region of the uterus
is located at the top of each image. Scale bars: 500 µm.
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Fig. 9. Invasive trophoblast cell distribution within day 11.5 uteroplacental
compartments of wild-type and Dprp-null mice exposed to normoxia or
hypobaric hypoxia. (A-C) Wild-type (+/+) and (D-F)
Dprp-null (-/-) mice were exposed to normoxia (A,D) or hypobaric
hypoxia (B,C,E,F). Trophoblast cells were identified by cytokeratin
immunostaining. C and F are high magnification images of the areas delineated
by the boxes in B and E, respectively. Note the decreased endovascular
trophoblast invasion (arrowheads in C,F) in the Dprp-null mice
exposed to hypoxia. The mesometrial region of the uterus is located at the top
of each image. Scale bars: 1 mm for A,B,D,E
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© The Company of Biologists Ltd 2007
