SRI Supplement to Reproductive Sciences - Volume 25 Number 1 - March 2018 - 157A

Scientific Abstracts

Reproductive Sciences Vol. 25, Supplement 1, March 2018

Table 1. Structural variability (number of branching points) in ex vivo perfused
human placenta.
Number of branching
points

mean ± SEM

Sections of cotyledon

PL1

N.A

52 ± 20

PL2

285

205

135

165 ± 53

PL3

183

105

109 ± 26

*Figure(s) will be available online.

T-143
Sex-Dependent Effect of Maternal High-Fat Diet on Endocannabinoid
System Expression in Rat Placenta. Raphael Mendonça†, Camilla DiasRocha†, Mariana Almeida†, Tania Ortiga-Carvalho*, Isis Trevenzoli*.
Federal University of Rio de Janeiro, Rio de Janeiro, Brazil.
INTRODUCTION: Maternal high-fat diet (HFD) consumption during
gestation is associated with placenta dysfunction and obesity programming
in the offspring. Obesity and HFD alter the expression and activity of the
endocannabinoid system (ECS) in a tissue dependent manner. The ECS
is mainly composed of the arachidonic acid-derived endocannabinoids,
anandamide and 2-araquidonoylglycerol; the cannabinoid receptors,
CB1 and CB2; and the metabolizing enzymes, FAAH and MAGL. ECS
is expressed in human and rat placenta, where endocannabinoids are
precursors for the increasing prostaglandin and prostanoid synthesis across
gestation. However, the effect of maternal HFD during preconceptional
and gestational periods on rat placenta ECS is unknown. We hypothesized
that maternal HFD would alter the content of CB1, CB2, FAAH and
MAGL in rat placenta at late gestation depending on fetal sex.
METHODS: Female rats received standard diet (C group; 9% fat) or
high-fat diet (HFD group; 28% fat) from 8 weeks before mating until
late gestation. Placentae were collected at gestational day 20.5 from male
and female fetuses. Placenta tissue was used for western blot analysis of
ECS components.
RESULTS: Maternal diet and fetal sex were considered as main
parameters for two-way ANOVA analysis. Maternal HFD increased CB1
and MAGL protein content only in male placenta (p<0.05), without effect
on female placenta. Interestingly, male rat placenta showed lower content
of CB1 and higher content of CB2, FAAH and MAGL compared with
female placenta (p<0.05).
CONCLUSION: Maternal HFD increased MAGL content in male
placenta, suggesting increased local production of arachidonic acid, which
may affect the n-6 lipid supply to male fetuses. Increased levels of n-6
lipids, such as arachidonic acid, is associated with increased levels of
CB1 in several tissues, as we observed in male placenta. In addition, we
previously showed that only HFD male pups present higher hypothalamic
levels of CB1 at birth, suggesting a relationship between placenta ECS
metabolism and expression of cannabinoid receptor in rat pups. The higher
levels of CB2, FAAH and MAGL found in male placenta, compared with
female placenta, suggests increased inflammation and arachidonic acid
levels in males and may be a mechanism underlying sex differences in
the developmental origins of health and disease (DOHaD).

T-144
The Use of Contrast-Enhanced Micro-CT for 3D Morphological
Assessment of the Mature Murine Placenta. Katrien De Clercq†,1
Eleonora Persoons†,1 Greet Kerckhofs*,2 Joris Vriens*.1 1KU Leuven,
Leuven, Belgium; 2UC Louvain, Louvain-la-Nueve, Belgium.
INTRODUCTION: The placenta plays an important role in normal
pregnancy and improper functioning can have detrimental consequences
leading to a broad range of pathological conditions, including intrauterine
growth restriction and fetal death. The placenta should therefore
be appropriately evaluated when placental defects are suspected.
Conventionally, the relative size of different placental layers is evaluated
by histologist evaluation. However, this technique is labor intensive and
subjected to tissue shrinkage, and it is difficult to draw conclusions with
regard to three-dimensional (3D) volumes when only two-dimensional
(2D) slides are being analysed.
METHODS: Contrast-enhanced microfocus computed tomography
(CE-CT) was validated as a fast, non-invasive, quantitative and robust
technique to evaluate simultaneously the placental morphology, the
volumes of the total placenta and its different layres, and the placental
vasculature at microscale level. The use of a noval, non-invasive contrast
agent, a Zirconium-substituted Keggin Polyoxometalate (Zr-POM),
allowed for a direct comparison between Zr-POM staining and (immuno-)
histological stainings in order to validate Zr-POM-based CE-CT.
RESULTS: Quantification of the placental volume by CE-CT indicated
an increase during Gestation, which could be attributed to a rise in volume
of the labyrinth. The volume of the junctional zone and the decidua
decreased. In addition, the substantial difference in cell morphology
between glycogen and spongiotrophoblast cells residing in the junctional
zone allowed for a quantification of their relative proportion. This
confirmed that the proportion of glycogen cells decreased towards term.
Moreover, the maternal central canal and its branching pattern could
be evaluated. Finally, as additional proof of concept, placentas of two
different mouse strains, C57BL6 and 129S6, were analyzed and confirmed
the differences in morphology as described in literature.
CONCLUSION: The use of CE-CT offers a novel, reliable and
quantitative method for a detailed 3D assessment of the placental
morphology and vasculature.

T-145
The Effects of Bacterial Lipopolysaccharide (LPS) Treatment of
Human Amnion Explants on Eicosanoid Secretion. Hassendrini N
Peiris†, Kanchan Vaswani, Fatema B Almughlliq, Yong Q Koh, Sarah
Reed, Murray D Mitchell*. University of Queensland, Brisbane, Australia.
INTRODUCTION: Prostaglandins (PGs) play important roles during
pregnancy and parturition in women. Arachidonic acid is converted into
PGs including PGE2, and PGF2α, and the latter metabolized into PGFM.
Endocannabinoids such as anandamide (AEA) can act as substrates
for enzymes of the prostaglandin biosynthetic pathways and can be
converted into related compounds such as prostamides (e.g. PGE2-EA,
PGF2α-EA). Mass spectrometry is being used to analyze prostaglandins
and prostamides with greater specificity and accuracy while providing the
ability to multiplex i.e. measure several analytes at once from the same
sample preparation. Aim: To measure prostaglandins and prostamides
(PGE2, PGF2α, PGFM, PGE2-EA and PGF2α-EA) secreted by amnion
explants upon treatment with an inflammatory stimulus (bacterial
lipopolysaccharide; LPS).
METHODS: Amnion tissues were collected from human term placentae
(n=6) collected at elective caesarean sections. Amnion tissue explants
were treated with LPS (5µg/mL) for 24 hrs. Explant culture media and
tissues were collected for subsequent analyses. Explant culture media and
standards were subjected to an extraction solution of methanol/formic
acid containing internal standards (PGE2-d4, PGF2α-d4, PGFM-d4, PGE2EA-d4 and PGF2α-EA-d4 250fmol each). Extracted samples and standards
were measured by mass spectrometry (LC-MS/MS).
RESULTS: Treatment with LPS resulted in an increased rate of
production of PGE2 (p< 0.0001) and PGF2α (p<0.05) compared to the basal
productions of these prostaglandins (control). No significant differences

Thursday Posters

specimens per cotyledon) were evaluated at 10X (Figure 1, red fluorescein
- fetal endothelium). Images were quantified using Image-Pro Premier
software (Media Cybernetics, Inc, Rockville, MD. USA) and Imaris 9
(Bitplane, USA). Number of branching points was calculated.
RESULTS: Volume of fetal capillaries ranged from 16.8% to 50% of
the volume of cotyledon and number of branching points was 31-285 per
specimen (Fig.1, Table 1). Fetal initial inflow pressure and flow-mediated
vasodilation (FMVD) did not correlate with these parameters.
CONCLUSION: Factors, other than vascular tree 3D composition, might
be responsible for the umbilical arterial resistance in vivo.

157A

Table of Contents for the Digital Edition of SRI Supplement to Reproductive Sciences - Volume 25 Number 1 - March 2018