Calcium signals from intact rabbit ciliary epithelium observed with confocal microscopy.

PURPOSE

We investigated patterns of evoked calcium signals to learn about the function of the calcium second messenger system in ciliary epithelium.

METHODS

Isolated infact ciliary processes were loaded with fluo-3/AM and observed with a Bio-Rad MRC-600 laser scanning confocal imaging system, before, during, and after perfusion with catecholamines, cholinergic agents, and autocoids.

RESULTS

One microM acetylcholine (ACH) and 10 microM carbachol (CARB) induced an atropine-sensitive increase in intracellular free calcium ion concentration ([Ca2+]i), considerably greater in NPE than in PE. 10 microM epinephrine (EPI) and 100 microM phenylephrine (PHE) increased [Ca2+]i in NPE and PE, in this case PE > NPE. These effects were blocked by prazosin. 10 mM caffeine (CAF) increased of [Ca2+]i in NPE and PE (NPE > PE) and sometimes produced very slow oscillations with an interval of 10 to 25 s. Prior administration of CAF strongly suppressed the effects of ACH, CARB, EPI, PHE, histamine, and adenosine 5-triphosphate (ATP). One hundred microM ryanodine (RYA) or thapsigargin (TG) increased [Ca2+]i in NPE and PE (NPE > PE).

CONCLUSIONS

In the ciliary epithelium: (1) different patterns of evoked transients and oscillations of calcium were seen in response to agonists; (2) NPE appeared to contain a predominance of muscarinic receptors, while the PE is dominated by alpha 1-adrenergic receptors and (3) the increase in [Ca2+]i by CAF or RYA or TG in either cell layer and the blocking effects of these agents upon agonist, evoking increases in [Ca2+]i, suggested involvement of both the cyclic adenosine diphosphate ribose and the inositol 1, 4, 5 triphosphate (InsP3) systems in the regulation of intracellular calcium.