Limits...
Local uterine Ang-(1-7) infusion augments the expression of cannabinoid receptors and differentially alters endocannabinoid metabolizing enzymes in the decidualized uterus of pseudopregnant rats.

Brosnihan KB, Pulgar VM, Gallagher PE, Neves LA, Yamaleyeva LM - Reprod. Biol. Endocrinol. (2015)

Bottom Line: CB1R mRNA was reduced by decidualization (2.7-fold, p < 0.001), but increased by Ang-(1-7) (1.9-fold, p < 0.05).CB2R mRNA was increased by decidualization (4-fold, p < 0.05) and by Ang-(1-7) (2.4-fold, p < 0.001).The enzyme metabolizing AEA, fatty acid amide hydrolase (FAAH), was reduced by decidualization (7.8 fold, p < 0.001) and unchanged by Ang-(1-7) (p > 0.05), whereas the enzyme metabolizing 2-arachidonoylglycerol, monoacyl glycerol lipase (MAGL), was unchanged by decidualization (p > 0.05) and increased by Ang-(1-7) (1.7 fold, p < 0.001).

View Article: PubMed Central - PubMed

Affiliation: Hypertension and Vascular Research Center, Wake Forest School of Medicine, Winston-Salem, NC, USA. bbrosnih@wakehealth.edu.

ABSTRACT

Background: Endocannabinoids (ECs) are important contributors to implantation and decidualization and are suppressed in early pregnancy. Elevated levels of anandamide (AEA), the endogenous ligand for the CB1 and CB2 receptors (R), interfere with receptivity of the blastocyst. Ang-(1-7) is down-regulated in the implantation site (IS) in normal pregnancy at day 7 of gestation. We determined the effects of intra-uterine angiotensin-(1-7) [Ang-(1-7)] (24 microg/kg/h) or vehicle given into the left uterine horn on the ECs in the decidualized uterus.

Methods: Ovariectomized rats were sensitized for the decidual cell reaction by steroid treatment and decidualization was induced by a bolus of oil injected into the left horn; the right horn served as a control.

Results: Decidualization increased endometrial permeability (3.1+/-0.2 vs. 7.1+/-0.5 uterus/muscle of cpm of (125)I-BSA, p < 0.0001). VEGF mRNA was increased by the decidualization (1.4-fold, p < 0.05) and by Ang-(1-7) (2.0-fold, p < 0.001). CB1R mRNA was reduced by decidualization (2.7-fold, p < 0.001), but increased by Ang-(1-7) (1.9-fold, p < 0.05). CB2R mRNA was increased by decidualization (4-fold, p < 0.05) and by Ang-(1-7) (2.4-fold, p < 0.001). The enzyme metabolizing AEA, fatty acid amide hydrolase (FAAH), was reduced by decidualization (7.8 fold, p < 0.001) and unchanged by Ang-(1-7) (p > 0.05), whereas the enzyme metabolizing 2-arachidonoylglycerol, monoacyl glycerol lipase (MAGL), was unchanged by decidualization (p > 0.05) and increased by Ang-(1-7) (1.7 fold, p < 0.001).

Conclusions: These findings report for the first time that Ang-(1-7) augments the expression of CB1R, CB2R and MAGL in the decidualized uterus and thus may interfere with the early events of decidualization.

Show MeSH
Time course of the sequence of hormone administration to ovariectomized rats and the timing of Ang-(1–7) or vehicle administration. Black areas represent periods of night time; numbers within light box areas indicate days relative to ovariectomy when hormones were injected subcutaneously. Arrows projecting from dose and type of hormone in boxes indicate the equivalent type of hormone that was administered sequentially. The time of day of injection is indicated. Decidualization induction and implantation of the miniosmotic pump occurred on day 5, and animals were sacrificed on day 10. E2 = 17-beta estradiol, P = progesterone.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4325957&req=5

Fig1: Time course of the sequence of hormone administration to ovariectomized rats and the timing of Ang-(1–7) or vehicle administration. Black areas represent periods of night time; numbers within light box areas indicate days relative to ovariectomy when hormones were injected subcutaneously. Arrows projecting from dose and type of hormone in boxes indicate the equivalent type of hormone that was administered sequentially. The time of day of injection is indicated. Decidualization induction and implantation of the miniosmotic pump occurred on day 5, and animals were sacrificed on day 10. E2 = 17-beta estradiol, P = progesterone.

Mentions: All procedures were approved by the Wake Forest School of Medicine Animal Care and Use Committee. Female Sprague–Dawley rats (n = 9-10/group) were obtained from Harlan Laboratories at 10 weeks of age and were ovariectomized under 2% isofluorane anesthesia. Five days after surgery animals were treated with a hormone regime [17-beta estradiol (0.1. 0.2, or 0.3 μg) and progesterone (1 or 4 mg)] as illustrated in Figure 1 and as described [8]. On day 5 as indicated on Figure 1, animals were anesthetized with 2% isoflurane and 0.1 ml of sesame oil was injected into the left uterine horn; an osmotic minipump (model 2ML2, pumping rate of 5 μL/hr) was placed in the left uterine horn for delivery of either 24 μg/kg/h of Ang-(1–7) in sterile phosphate sodium buffer (PBS, pH 7.4) or vehicle. PE60 tubing attached to the minipump was inserted into the uterus lumen until it reached a plastic cuff placed 1 mm from the tip of the catheter. Sutures secured the catheter (before and after the cuff) and either Ang-(1–7) or PBS was infused. The right horn was not injected or infused and served as a control. After five days of treatment, animals were euthanized by decapitation and trunk blood was collected in a cocktail of inhibitors as previously described [9]. The non-infused and infused uterine horns were removed, weighed, snap-frozen on dry ice for mRNA analysis, or fixed in 10% neutral buffered formalin solution for immunostaining.Figure 1


Local uterine Ang-(1-7) infusion augments the expression of cannabinoid receptors and differentially alters endocannabinoid metabolizing enzymes in the decidualized uterus of pseudopregnant rats.

Brosnihan KB, Pulgar VM, Gallagher PE, Neves LA, Yamaleyeva LM - Reprod. Biol. Endocrinol. (2015)

Time course of the sequence of hormone administration to ovariectomized rats and the timing of Ang-(1–7) or vehicle administration. Black areas represent periods of night time; numbers within light box areas indicate days relative to ovariectomy when hormones were injected subcutaneously. Arrows projecting from dose and type of hormone in boxes indicate the equivalent type of hormone that was administered sequentially. The time of day of injection is indicated. Decidualization induction and implantation of the miniosmotic pump occurred on day 5, and animals were sacrificed on day 10. E2 = 17-beta estradiol, P = progesterone.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4325957&req=5

Fig1: Time course of the sequence of hormone administration to ovariectomized rats and the timing of Ang-(1–7) or vehicle administration. Black areas represent periods of night time; numbers within light box areas indicate days relative to ovariectomy when hormones were injected subcutaneously. Arrows projecting from dose and type of hormone in boxes indicate the equivalent type of hormone that was administered sequentially. The time of day of injection is indicated. Decidualization induction and implantation of the miniosmotic pump occurred on day 5, and animals were sacrificed on day 10. E2 = 17-beta estradiol, P = progesterone.
Mentions: All procedures were approved by the Wake Forest School of Medicine Animal Care and Use Committee. Female Sprague–Dawley rats (n = 9-10/group) were obtained from Harlan Laboratories at 10 weeks of age and were ovariectomized under 2% isofluorane anesthesia. Five days after surgery animals were treated with a hormone regime [17-beta estradiol (0.1. 0.2, or 0.3 μg) and progesterone (1 or 4 mg)] as illustrated in Figure 1 and as described [8]. On day 5 as indicated on Figure 1, animals were anesthetized with 2% isoflurane and 0.1 ml of sesame oil was injected into the left uterine horn; an osmotic minipump (model 2ML2, pumping rate of 5 μL/hr) was placed in the left uterine horn for delivery of either 24 μg/kg/h of Ang-(1–7) in sterile phosphate sodium buffer (PBS, pH 7.4) or vehicle. PE60 tubing attached to the minipump was inserted into the uterus lumen until it reached a plastic cuff placed 1 mm from the tip of the catheter. Sutures secured the catheter (before and after the cuff) and either Ang-(1–7) or PBS was infused. The right horn was not injected or infused and served as a control. After five days of treatment, animals were euthanized by decapitation and trunk blood was collected in a cocktail of inhibitors as previously described [9]. The non-infused and infused uterine horns were removed, weighed, snap-frozen on dry ice for mRNA analysis, or fixed in 10% neutral buffered formalin solution for immunostaining.Figure 1

Bottom Line: CB1R mRNA was reduced by decidualization (2.7-fold, p < 0.001), but increased by Ang-(1-7) (1.9-fold, p < 0.05).CB2R mRNA was increased by decidualization (4-fold, p < 0.05) and by Ang-(1-7) (2.4-fold, p < 0.001).The enzyme metabolizing AEA, fatty acid amide hydrolase (FAAH), was reduced by decidualization (7.8 fold, p < 0.001) and unchanged by Ang-(1-7) (p > 0.05), whereas the enzyme metabolizing 2-arachidonoylglycerol, monoacyl glycerol lipase (MAGL), was unchanged by decidualization (p > 0.05) and increased by Ang-(1-7) (1.7 fold, p < 0.001).

View Article: PubMed Central - PubMed

Affiliation: Hypertension and Vascular Research Center, Wake Forest School of Medicine, Winston-Salem, NC, USA. bbrosnih@wakehealth.edu.

ABSTRACT

Background: Endocannabinoids (ECs) are important contributors to implantation and decidualization and are suppressed in early pregnancy. Elevated levels of anandamide (AEA), the endogenous ligand for the CB1 and CB2 receptors (R), interfere with receptivity of the blastocyst. Ang-(1-7) is down-regulated in the implantation site (IS) in normal pregnancy at day 7 of gestation. We determined the effects of intra-uterine angiotensin-(1-7) [Ang-(1-7)] (24 microg/kg/h) or vehicle given into the left uterine horn on the ECs in the decidualized uterus.

Methods: Ovariectomized rats were sensitized for the decidual cell reaction by steroid treatment and decidualization was induced by a bolus of oil injected into the left horn; the right horn served as a control.

Results: Decidualization increased endometrial permeability (3.1+/-0.2 vs. 7.1+/-0.5 uterus/muscle of cpm of (125)I-BSA, p < 0.0001). VEGF mRNA was increased by the decidualization (1.4-fold, p < 0.05) and by Ang-(1-7) (2.0-fold, p < 0.001). CB1R mRNA was reduced by decidualization (2.7-fold, p < 0.001), but increased by Ang-(1-7) (1.9-fold, p < 0.05). CB2R mRNA was increased by decidualization (4-fold, p < 0.05) and by Ang-(1-7) (2.4-fold, p < 0.001). The enzyme metabolizing AEA, fatty acid amide hydrolase (FAAH), was reduced by decidualization (7.8 fold, p < 0.001) and unchanged by Ang-(1-7) (p > 0.05), whereas the enzyme metabolizing 2-arachidonoylglycerol, monoacyl glycerol lipase (MAGL), was unchanged by decidualization (p > 0.05) and increased by Ang-(1-7) (1.7 fold, p < 0.001).

Conclusions: These findings report for the first time that Ang-(1-7) augments the expression of CB1R, CB2R and MAGL in the decidualized uterus and thus may interfere with the early events of decidualization.

Show MeSH