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CB1R is expressed in fetal endothelium, cytotrophoblast and

syncytiotrophoblast (Figures 5, 6). The expression of CB1R protein was

decreased in the villous tree of HFD animals (214.11 ± 1.28 vs 209.22 ±

0.98; CTR vs. HFD, respectively, p=0.05, Mann-Whitney U test). The

placental (203.00 ± 12.49 g vs. 226.23 ± 7.13 g) and fetal (810 ± 17.76

g vs 849. 59 ± 24.47 g) weights did not differ between the groups

(Figure 7).

GENDER-SPECIFIC PLACENTAL EXPRESSION OF CANNABINOID RECEPTOR ONE (CB1R)

IN BABOON MODEL OF OBESITY

Introduction

Maira Carrillo1, Cun Li2,3, Cezary Skobowiat4,5, Gary Ventolini1, Marcel Chuecos1, Patrick Joseph6, Edward Dick7, Gene Hubbard8,

Peter Nathanielsz2,3, Natalia Schlabritz-Loutsevitch1

1 Dept. of Obstetrics and Gynecology, Texas Tech University Health Science Center at the Permian Basin, Odessa, TX, USA ; 2 Texas Pregnancy and Life Course Health Center, Texas Biomedical Research Institute, San Antonio, TX, USA; 3 Dept. of Animal Science, University of Wyoming, Laramie, WY, USA;, 4 Dept. of Pharmacodynamics and Molecular Pharmacology, Collegium Medicum Nicolaus Copernicus University of Poland, Torun, Poland; 5 Dept. of Dermatology, University of Alabama at Birmingham, Birmingham, AL, USA,

6 Dept. of Pathology, University of Tennessee Health Sciences Center, Memphis, TN, USA;7 Texas Biomedical Research Institute, San Antonio, TX, USA; 8 Dept. of Pathology, University of Texas Health Science Center at San Antonio, San Antonio, TX, USA

To explore the role of dietary fat composition (higher fat/higher

carbohydrate diet) vs. overeating pattern in placental CB1R

expression.

Objectives

Baboons (Papio spp) were fed a diet of 45% fat (HFD; n=4) while

controls (CTR; n=4) ate 12% fat from at least 9 months prior to

conception through pregnancy until 0.9 gestation (Placenta 2009

30(9):760). Immunohistochemistry (IHC) was performed on placental

tissue of male fetuses from mothers of HFD and control groups (Figure

5,6) using the CB1R monoclonal primary antibody (Immunogenes;

Budakeszi, Hungary), the secondary antibody is included in the

Vectastain ABC kit (Vector laboratories; Burlingame, CA); methodology

was performed according to manufacturers instructions. The slides were

scanned using the NanoZoomer SQ (Hamamatsu; Middlesex, NJ),

quantification was performed using ImageScope™ v11.1.2.752 by

Aperio (Leica Biosystems; Buffalo Grove, IL). Conclusion

Availability of the cytoplasmic pool for placental CB1R is regulated by a

high fat diet, but not increased food intake in this baboon model of

maternal obesity. Decreased placental CBR1 availability might alter

eCBs regulated fetal vascular reactivity

Acknowledgement Authors thank personnel of the Texas Biomedical Institute and Center for Pregnancy and Newborn Research (UTHSC—San Antonio) for their help. This study was partially supported by Texas

Biomedical Research Institute Grant C06 RR013556 and NIH grant HD21350 to Dr. Peter Nathanielsz (UTHSC—San Antonio), UTHSCSA ERC New Investigator Award to N.S.-L., NIH NCRR

grant P51 RR013986 to the Southwest National Primate Research. This work was supported by the TTUHSC start-up funds to N.S-L., Dr. Carrillo’s travel was supported by the NIH young

investigator award. Also supported NIH R24OD010916 and P51 RR013986.

Results

Methods

Maternal obesity is associated with offspring behavioral and

cardiovascular developmental problems through poorly understood

mechanisms. Endocannabinoids (eCB) are lipid derivatives, linking

metabolism to mental and cardiovascular health (Figure 1). CB1R – a

G-protein-coupled receptor- is the main target of endogenous (2-

arachidonoylglycerol (2-AG) and N-arachidonoylethanolamine (AEA))

and exogenous cannabinoids (e.g. marijuana). We previously reported

a seemingly paradoxical decrease of CB1R ligands’ concentrations

(Figure 2), decreased mRNA, but not CB1R protein expression in

placentas of obese baboons (increased food consumption model)

(Placenta 2013 34(11):983) (Figure 3) and decreased placental protein

expression of CB1R and CB2R in obese pregnant women carrying

male fetuses (Figure 4.) (AJOG, 2014, 210(1):S94).

References

1. Brocato B, Zoerner AA, Janjetovic Z, Skobowiat C, Gupta S, Moore BM 2nd, Slominski A, Zhang J, Schenone

M, Phinehas R, Ferry RJ Jr, Dick E Jr, Hubbard GB, Mari G, Schlabritz-Loutsevitch N. Endocannabinoid

crosstalk between placenta and maternal fat in a baboon model (Papio spp.) of obesity..Placenta. 2013

Nov;34(11):983-9. doi: 10.1016/j.placenta.2013.08.007.

2. Farley D, Tejero ME, Comuzzie AG, Higgins PB, Cox L, Werner SL, Jenkins SL, Li C, Choi J, Dick EJ Jr,

Hubbard GB, Frost P, Dudley DJ, Ballesteros B, Wu G, Nathanielsz PW, Schlabritz-Loutsevitch NE. Placenta.

Feto-placental adaptations to maternal obesity in the baboon. 2009 Sep;30(9):752-60. doi:

10.1016/j.placenta.2009.06.007. Epub 2009 Jul 25.

3. Maccarrone M, Bab I, Bíró T, Cabral GA, Dey SK, Di Marzo V, Konje J, Kunos G, Mechoulam, Pacher P,

Sharkey KA, Zimmer A .Endocannabinoid signaling at the periphery: 50 years after THC. Trends Pharmacol Sci.

2015 May;36(5):277-96. doi: 10.1016/j.tips.2015.02.008. Epub 2015 Mar 18

4. N. Schlabritz-Loutsevitch, J. Samson, S. Gupta, M.Schenone, . Brocato, D.e Tate, R.Krutilina, C.Skobowiat, A.

Slominski, B.Moore, G. Mari. Fetal Gender-Specific Placental “Endocannabinoidome” in Maternal Obesity. 2014

American Journal of Obstetrics and Gynecology, Supplement to Vol. 210, N1,S. 94

5. F. Xavier Pi-Sunyer, MD, MPH. Role of the Endocannabinoid System in Contributing to Obesity. 2008.

http://www.medscape.org/viewarticle/567416

Weight of male fetuses delivered by control

(n=4) and HFD (n=4) dams.

HFD Control Negative control

Figure 5. CB1R proteins expression in placenta of male fetuses in HFD (A) and control (B,C).

Black arrow shows the syncytiotrophoblast, green arrow shows cytotrophoblast (A), and yellow

arrow shows endothelium (A, B, and C).

Placental CB1R expression in HFD (n=4)

and non-obese (n=4) dams, carrying

male fetus

Weight of placenta in HFD (n=4) and

non-obese (n=4) dams, carrying

male fetuses.

A B C Figure 2. Concentrations of AEA and 2AG in placenta, maternal serum and maternal fat of

obese (n=4) and non-obese (n=4) pregnant baboons (Placenta 2013 34(11):983).

Figure 3. CB1R protein expression and quantifications (A,B) and mRNA expression (C) in

placenta of naturally obese (n=4) and non-obese (n=4) baboons. (Placenta 2013 34(11):983).

A B C

Figure 4. Placental protein expression of CB1R and CB2R in obese (n=3) and non-obese (n=4)

pregnant women carrying male fetuses (A). Quantification and analysis by western blot (B)

(AJOG, 2014, 210(1):S94).

A B

Figure 7. Placental CB1R protein expression in

HFD (n=4) and non-obese (n=4) dams (A);

placental (B) and fetal weights (C).

Figure 6. CB1R protein expression (A) and H&E staining (B) in placenta of male fetuses, control

group. Black arrow is syncytiotrophoblast, green arrow is cytotrophoblast and yellow arrow

endothelium.

A B C

Figure 1. Endocannabinoid system and the effects on various systems of the body. (F. Xavier Pi-

Sunyer, 2008)

A B

B

C

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A