Drug-dependent Ca²⁺ mobilization in organ-cultured rabbit ciliary processes

Abstract

This study was conducted to determine whether drug-dependent changes in cytosolic Ca²⁺ concentration take place in the ciliary nonpigment epithelial cells of rabbits under more physiological conditions. Iris-ciliary body from pigmented rabbits in organ-culture was loaded with a Ca²⁺−sensitive fluorescent dye, fura-2, and a video-imaging system with an image analyzer was employed. Using this method fluorescence from nonpigmented epithelial cells can be analyzed without interference from fluorescence from pigmented ciliary epithelial cells. Among the drugs studied, norepinephrine and carbachol induced Ca²⁺ transients in the nonpigmented epithelial cells of organ-cultured ciliary processes. Epinephrine, isoproterenol, dopamine, neuropeptide Y, and substance P at the concentration of 10⁻⁶ to 10⁻³ M failed to elicit a response. The cytosolic free Ca²⁺ concentration of the cells in the resting state, as determined by an in vitro calibration curve, was 166 nM. The peak free cytosolic Ca²⁺ concentration induced by norepinephrine was about 263 nM, and that induced by carbachol was more than 1,000 nM. The carbachol-induced response was larger in magnitude and longer in duration than that induced by norepinephrine. Not uncommonly, the carbachol-induced response lasted more than 15 min. The response was diminished in both peak height and duration by chelation of extracellular Ca²⁺. Atropine abolished the response showing the response being mediated by a muscarinic receptor.

Key Words:

• adrenergic
• cholinergic drugs
• Ca²⁺ mobilization
• ciliary process
• organ culture
• rabbit