2-(4-(2-carboxyethyl)phenethylamino)-5--n-ethylcarboxamidoadenosine and adenosine-amine-congener

Intraocular pressure (IOP) is regulated by the resistance to outflow of the eye's aqueous humor. Elevated resistance raises IOP and can cause glaucoma. Despite the importance of outflow resistance, its site and regulation are unclear. The small size, complex geometry, and relative inaccessibility of the outflow pathway have limited study to whole animal, whole eye, or anterior-segment preparations, or isolated cells. We now report measuring elemental contents of the heterogeneous cell types within the intact human trabecular outflow pathway using electron-probe X-ray microanalysis. Baseline contents of Na(+), K(+), Cl(-), and P and volume (monitored as Na+K contents) were comparable to those of epithelial cells previously studied. Elemental contents and volume were altered by ouabain to block Na(+)-K(+)-activated ATPase and by hypotonicity to trigger a regulatory volume decrease (RVD). Previous results with isolated trabecular meshwork (TM) cells had disagreed whether TM cells express an RVD. In the intact tissue, we found that all cells, including TM cells, displayed a regulatory solute release consistent with an RVD. Selective agonists of A(1) and A(2) adenosine receptors (ARs), which exert opposite effects on IOP, produced similar effects on juxtacanalicular (JCT) cells, previously inaccessible to functional study, but not on Schlemm's canal cells that adjoin the JCT. The results obtained with hypotonicity and AR agonists indicate the potential of this approach to dissect physiological mechanisms in an area that is extremely difficult to study functionally and demonstrate the utility of electron microprobe analysis in studying the cellular physiology of the human trabecular outflow pathway in situ.

Topics: Adenosine; Adenosine A1 Receptor Agonists; Adenosine A2 Receptor Agonists; Aqueous Humor; Cell Size; Chlorides; Electron Probe Microanalysis; Enzyme Inhibitors; Feasibility Studies; Humans; Hypotonic Solutions; Intraocular Pressure; Norbornanes; Osmotic Pressure; Ouabain; Phenethylamines; Phosphorus; Potassium; Receptor, Adenosine A1; Receptors, Adenosine A2; Sodium; Sodium-Potassium-Exchanging ATPase; Trabecular Meshwork

In isolated guinea pig hearts saline perfused at constant flow, adenosine A(1), A(2A), and A(3) (A(x)) agonists covalently bound to a large polymer (Pol; 2,000 kDa) were intracoronarily administered, and three effects were studied: dromotropic, vascular and inotropic. The rank order of potencies were the following: dromotropic (Pol-A(2A)Pol-A(1)>Pol-A(3)) and vascular and inotropic (Pol-A(2A)> or =Pol-A(1)Pol-A(3)), where the rank order of potency for Pol-A(x) depends on the part of the coronary vascular network involved; i.e., there is a vascular heterogeneity. The large size of Pol-A(x) prevents extravascular diffusion and causes it to act solely in the endothelial luminal surface. This implies their cardiac effects are due to endothelial mediators. Inhibition of nitric oxide (NO) and prostaglandin (PG) synthesis with N(G)-nitro-l-arginine methyl ester and indomethacin, respectively, show that the three cardiac effects of Pol-A(1) were mediated by NO and PG, whereas for Pol-A(2A) and Pol-A(3) the mediator was mainly NO but not PG. These results suggest that if Pol-A(x) activated the corresponding endothelial A(x)-receptor subtype, a different mediator would be produced.

Topics: Adenosine; Animals; Aorta; Coronary Vessels; Cyclooxygenase Inhibitors; Dose-Response Relationship, Drug; Enzyme Inhibitors; Guinea Pigs; Heart; In Vitro Techniques; Indomethacin; Male; NG-Nitroarginine Methyl Ester; Perfusion; Phenethylamines; Polymers; Protein Isoforms; Purinergic P1 Receptor Agonists; Receptors, Purinergic P1; Vasoconstriction; Vasodilation

A previous study reported that beta-endorphin and morphine administered supraspinally produce antinociception by activating different descending pain inhibitory systems. The present study was designed to investigate the blocking effects of A1 or A2 adenosine receptors in the spinal cord on antinociception induced by supraspinally administered mu- and epsilon-opioid receptor agonists. The effects of 1,3-dipropyl-8-(2-amino-4-chloro-phenyl)-xanthine (PACPX; an A1 adenosine receptor antagonist) or 3,7-dimethyl-1-propargylxanthine (DMPX; an A2 adenosine receptor antagonist) on the antinociception induced by morphine (a mu-opioid receptor agonist) or beta-endorphin (an epsilon-opioid receptor agonist) administered intracerebroventricularly (i.c.v.) were studied. The antinociception was assayed by the tail-flick test. DMPX at doses of 1-40 micrograms (which administered intrathecally alone did not affect the latencies of tail-flick thresholds), attenuated dose-dependently the inhibition of the tail-flick response induced by i.c.v. administered morphine (0.5 microgram) or beta-endorphin (1 microgram). PACPX at doses of 1-40 micrograms (which administered intrathecally alone did not affect the latencies of tail-flick thresholds), attenuated dose-dependently the inhibition of the tail-flick response induced by i.c.v. administered beta-endorphin but not morphine. These results suggest that A2 but not A1 adenosine receptors in the spinal cord may be involved in the antinociception induced by supraspinally administered morphine, while the antinociception induced by supraspinally administered beta-endorphin appears to be mediated by spinal A1 and A2 adenosine receptors. These results support the hypothesis that morphine and beta-endorphin administered supraspinally produce antinociception by different neuronal mechanisms.

Topics: Adenosine; Analgesics; Animals; beta-Endorphin; Dose-Response Relationship, Drug; Injections, Intraventricular; Injections, Spinal; Male; Mice; Mice, Inbred ICR; Morphine; Phenethylamines; Purinergic P1 Receptor Antagonists; Reaction Time; Theobromine; Xanthines