benzofurans and pyridoxal-phosphate-6-azophenyl-2--4--disulfonic-acid

To clarify the basic mechanism involved in the pathophysiology of cystitis by characterizing the urodynamic parameters, pharmacologically relevant (muscarinic and purinergic) receptors, and the in vivo release of adenosine triphosphate (ATP) in the bladder of hydrochloric acid (HCl)-treated rats.. The muscarinic and purinergic receptors in rat tissue were measured by radioreceptor assays using (N-methyl-³H) scopolamine methyl chloride ([³H]NMS) and αβ-methylene-ATP (2,8-³H) tetrasodium salt ([³H]αβ-MeATP), respectively. The urodynamic parameters and ATP levels were measured using a cystometric method and the luciferin-luciferase assay, respectively.. In the HCl-treated rats, the micturition interval and micturition volume were significantly (48% and 55%, respectively, P <.05) decreased and the number of micturitions was significantly (3.2-fold, P <.05) increased compared with those of the control rats. The maximal number of binding sites for [³H]NMS and [³H]αβ-MeATP was significantly (55% and 72%, respectively, P <.001) decreased in the bladder of HCl-treated rats, suggesting downregulation of both muscarinic and purinergic receptors. In the HCl-treated rats, the inhibition constant, K(i), values for oxybutynin, solifenacin, and darifenacin were significantly (1.3-1.4-fold, P <.05) increased, but those for tolterodine and AF-DX116 were unchanged. Similarly, the inhibition constant for A-317491, pyridoxal-phosphate-6-azophenyl-2',4'-disulfonic acid tetrasodium, and MRS2273 was significantly (5.5, 11, and 7.6-fold, respectively, P <.001) increased. Furthermore, the in vivo release of ATP was significantly (P <.05) enhanced in the HCl-treated rat bladder.. Both muscarinic and purinergic mechanisms might be, at least in part, associated with the urinary dysfunction due to cystitis.

Topics: Adenosine Triphosphate; Animals; Benzhydryl Compounds; Benzofurans; Cresols; Cystitis; Disease Models, Animal; Down-Regulation; Female; Hydrochloric Acid; Mandelic Acids; N-Methylscopolamine; Organophosphonates; Phenols; Phenylpropanolamine; Pirenzepine; Polycyclic Compounds; Pyridoxal Phosphate; Pyrrolidines; Quinuclidines; Radioligand Assay; Rats; Rats, Sprague-Dawley; Receptors, Muscarinic; Receptors, Purinergic; Solifenacin Succinate; Tetrahydroisoquinolines; Tolterodine Tartrate; Urinary Bladder; Urination; Urodynamics

Here we show that rabbit Müller cell differentiation from radial glial progenitor cells is accompanied by a decreasing capability to respond to specific stimuli (depolarization and extracellular adenosine 5'-triphosphate [ATP]) with an elevation of intracellular calcium. Intracellular free calcium was recorded in retinal wholemounts from young (postnatal days [P] 2 to 31) and adult rabbits. Images were taken from the nerve fiber/ganglion cell layers where the endfeet of radial glial/ Müller cells can be identified after selective uptake of calcium-sensitive dyes. The area of responding endfeet was determined as the percentage of the total area occupied by Müller cell endfeet, as an estimate of the percentage of responding cells. In response to depolarization (50 mM potassium), an increase of intracellular free calcium occurred in 19% of cells from young postnatal retinae (P2-31) but only in 2% from adults. This depolarization-induced calcium rise was caused both by a calcium influx from extracellular space and by an intracellular calcium release. The latter response was inhibited by the P2 receptor blocker pyridoxal phosphate 6-azophenyl-2',4'-disulfonic acid (PPADS), indicating that extracellular calcium-independent ATP release into the extracellular space occurs during retinal depolarization. When extracellular ATP (200 microM) was applied, calcium responses were recorded in 83% of cells from young postnatal retinae (P2-6); in the course of further development, both the percentage of responding cells (7% in retinae from adult rabbits) and the amplitude of the calcium responses decreased. It is concluded that during the differentiation of immature radial glia into mature Müller cells, stimulus-evoked intracellular calcium signaling mechanisms change.

Topics: Adenosine; Adenosine Triphosphate; Aging; Animals; Benzofurans; Calcium; Calcium Signaling; Cell Count; Cell Differentiation; Cell Size; Fluorescent Dyes; Imidazoles; In Vitro Techniques; Magnesium; Neuroglia; Potassium; Pyridoxal Phosphate; Rabbits; Retina; Stem Cells