Real-Time Intracellular Ca2+ Mobilization by Travoprost Acid, Bimatoprost, Unoprostone, and Other Analogs via Endogenous Mouse, Rat, and Cloned Human FP Prostaglandin Receptors

Abstract

The ability of a number of prostaglandin F 2␣ (PGF 2␣) . In FP functional studies, travoprost acid (EC 50 ϭ 17.5-37 nM, n ϭ 13), bimatoprost acid (EC 50 ϭ 23.3-49.0 nM, n ϭ 6 -12), unoprostone (EC 50 ϭ 306-1270 nM, n ϭ 4 -8), bimatoprost (EC 50 ϭ 3070-3940 nM, n ϭ 4 -9), and Lumigan (EC 50 ϭ 1470 -3190 nM, n ϭ 5-9) concentration dependently stimulated [Ca 2ϩ ] i mobilization via the rat (A7r5 cells), mouse (3T3 cells), and cloned human ocular FP prostanoid receptors. The rank order of potency of these compounds at the FP receptor of the three species was similar and in good agreement with the determined binding affinities. The agonist effects of these compounds were concentration dependently blocked by the FP receptor-selective antagonist, AL-8810 (11␤-fluoro-15-epi-15-indanyl-tetranor PGF 2␣ ) (K i ϭ 0.6 -1.3 M). These studies have demonstrated that bimatoprost, unoprostone, and bimatoprost acid possess direct agonist activities at the rat, mouse, and human FP prostanoid receptor and that travoprost acid is the most potent of the synthetic FP prostaglandin analogs tested.

Characterization of a Rabbit Kidney Prostaglandin F2α Receptor Exhibiting Gi-restricted Signaling That Inhibits Water Absorption in the Collecting Duct

“…Like PGE 2 , latanoprost significantly blunts vasopressin-stimulated osmotic water permeability in the rabbit CCD; however, unlike PGE 2 , latanoprost does not increase intracellular calcium in this segment (15,20). This latter result was unanticipated because previous studies in cultured smooth muscle cells and fibroblasts demonstrate that FP receptor activation increases intracellular calcium and inositol trisphosphate production (32,34,37). Nevertheless, significant differences exist between the present studies and those previous studies, including the cell type tested (e.g.…”

PGF2α-associated vascular smooth muscle hypertrophy is ROS dependent and involves the activation of mTOR, p70S6k, and PTEN

“…After reaching 80-90% confluence, cells were made quiescent by incubation in serum-free DMEM for 48 h. In the evaluation of intracellular signaling studies, cells were stimulated with agonist (1 μM (+)-fluprostenol) or vehicle (ethanol) at 37°C in serum-free DMEM for 15 or 30 min. This concentration of fluprostenol is well above the K i and is consistent with that used by other investigations (14,25). For inhibition studies and protein accretion studies, A7r5 cells were seeded in 6-well plates (2.5× 10 5 cells/well) in DMEM supplemented with 10% FBS and antibiotics.…”

Pharmacological Evidence for a Functional Serotonin-2B Receptor in a Human Uterine Smooth Muscle Cell Line

“…Intracellular calcium mobilization [Ca 2ϩ ] i induced by 5-HT and other serotonergic compounds in HUSMCs was studied using a FLIPR instrument (Molecular Devices) (Schroeder and Neagle, 1996). Evaluation of the functional agonist activity of test compounds was performed using a protocol described previously (Kelly et al, 2003;May et al, 2003). In brief, confluent cell monolayers of human uterine smooth muscle cells were trypsinized, pelleted, and seeded at a density of 20,000 cells per well in black-walled, 96-well tissue culture plates and grown to confluence.…”