selective antagonist of the ep receptors ep2  (Tocris)

Journal: American Journal of Physiology - Heart and Circulatory Physiology

Article Title: A novel role for PGE 2 -EP 4 in the developmental programming of the mouse ductus arteriosus: consequences for vessel maturation and function

doi: 10.1152/ajpheart.00294.2023

Figure Lengend Snippet: Drug treatment protocols. Pregnant dams were treated with selective antagonists to the prostaglandin E receptors EP 1 (SC-51322), EP 2 (pF-04418948), EP 3 (L798,106), and EP 4 (L161,982, ONO-AE3-208), a selective agonist to EP 4 (TCS2510), prostaglandin E 2 (PGE 2 ), or an inhibitor of nitric oxide (NO) synthase N G -nitro- l -arginine methyl ester ( l -NAME) on the indicated days of gestation [ day 1 ( D1 ) = presence of vaginal plug]. Fetal exposure studies: Protocol A examined the effect of acute EP receptor antagonism on the in utero ductus arteriosus (DA), with DA scoring 4 h after maternal intraperitoneal administration. Neonatal injection studies: Protocol B examined the effect of postnatal EP receptor stimulation on DA constriction, with drugs being administered to offspring 30 min after delivery and DA scoring 4 h later. Maternal gavage studies: Protocol C examined how inhibition of EP 4 or NO synthase affected fetal DA patency on select days of pregnancy. Drugs were administered the morning of the indicated day and DA patency assessed 4 h later. Chronic gavage studies: Protocol D examined the effect of chronically inhibiting EP 4 signaling over a defined window of pregnancy. Mice were delivered via cesarean section on the morning of D19 with their DAs scored 4 h later. This approach was repeated during discrete windows in Protocols E, F, and G . PP1, postpartum day 1 .

Article Snippet: In protocol A , selective antagonists of the EP receptors EP [SC-51322, 10 mg/kg/dose once ( , ); ip; TOCRIS], EP 2 [PF-04418948, 10 mg/kg/dose once; ip ( ); TOCRIS], EP 3 [L-798,106, 5 mg/kg/dose once ( ); ip; TOCRIS], and EP 4 [L-161,982, 100 mg/kg/dose once hourly, 4 total ( ) (originally 10 mg/kg/dose once, one total, but increased to 4 total doses because of short in vivo half-life); ip; TOCRIS; or AE3-208, 10 mg/kg/dose once; ip; ONO Pharmaceuticals] ( , ) were administered to CD1 WT dams at 0800 h on the morning of D19 .

Techniques: In Utero, Injection, Inhibition

Journal: American Journal of Physiology - Heart and Circulatory Physiology

Article Title: A novel role for PGE 2 -EP 4 in the developmental programming of the mouse ductus arteriosus: consequences for vessel maturation and function

doi: 10.1152/ajpheart.00294.2023

Figure Lengend Snippet: Prostaglandin E 2 (PGE 2 ) receptor EP 4 is the predominant EP receptor in the mouse ductus arteriosus (DA). A : time course of EP receptor expression in the DA. Ptger4 expression increased with advancing gestation compared with day 15 ( D15 ) ( P < 0.05; Kruskal–Wallis) and was significantly greater than individual EP subtypes at each gestational stage (* P < 0.05; Kruskal–Wallis) ( n = 3 biological replicates). B : localization of Ptger4 (EP 4 ) expression in the DA and outflow tracts. C : cannulated ex vivo preparation of D19 CD1 wild-type (WT) DA for vessel myography. D and E : representative tracing of a PGE 2 concentration response curve (CRC; D ) and cumulative response curves of EP 4 -inhibited and control DAs following O 2 -induced preconstriction ( E ), demonstrating the potent and EP 4 -predominant effects of PGE 2 on the isolated DA. F : in vivo studies demonstrate a shift in neonatal DA patency rates in response to injections of the selective EP 4 agonist TCS2510, resulting in patent DA (PDA). G : representative images of DA patency in response to selective EP receptor antagonists, scored on a 5-point noncontinuous scale, showing 100% patency ( top ) and 0% patency ( bottom ). H : in utero exposure to selective EP receptor antagonists resulted in fetal DA constriction in response to 2 selective EP 4 antagonists, but not to EP 1 EP 2 , or EP 3 antagonists. AE3-208 was used in subsequent EP 4 inhibitor studies because of its increased potency and DA effects. **** P < 0.001 compared with control ( E ) or vehicle ( F and H ) ( A , Kruskal–Wallis; E , two-way ANOVA; F and H , χ 2 ). aAo, ascending aorta; BL, baseline; bPA, branch pulmonary artery; dAo, descending aorta; NS, not significant; PA, pulmonary artery.

Article Snippet: In protocol A , selective antagonists of the EP receptors EP [SC-51322, 10 mg/kg/dose once ( , ); ip; TOCRIS], EP 2 [PF-04418948, 10 mg/kg/dose once; ip ( ); TOCRIS], EP 3 [L-798,106, 5 mg/kg/dose once ( ); ip; TOCRIS], and EP 4 [L-161,982, 100 mg/kg/dose once hourly, 4 total ( ) (originally 10 mg/kg/dose once, one total, but increased to 4 total doses because of short in vivo half-life); ip; TOCRIS; or AE3-208, 10 mg/kg/dose once; ip; ONO Pharmaceuticals] ( , ) were administered to CD1 WT dams at 0800 h on the morning of D19 .

Techniques: Expressing, Ex Vivo, Concentration Assay, Control, Isolation, In Vivo, In Utero