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Roles of Calcium, PKA, and PKG

Orna Zagoory, Alex Braiman and Zvi Priel

Department of Chemistry, Ben-Gurion University of the Negev, Beer-Sheva 84105, Israel

Address correspondence to Dr. Z. Priel, Department of Chemistry, Ben-Gurion University of the Negev, P.O. Box 653, Beer-Sheva 84105, Israel. Fax: 972-8-6900046; E-mail: alon{at}bgumail.bgu.ac.il

Stimulation of ciliary cells through muscarinic receptors leads to a strong biphasic enhancement of ciliary beat frequency (CBF). The main goal of this work is to delineate the chain of molecular events that lead to the enhancement of CBF induced by acetylcholine (ACh). Here we show that the Ca²⁺, cGMP, and cAMP signaling pathways are intimately interconnected in the process of cholinergic ciliary stimulation. ACh induces profound time-dependent increase in cGMP and cAMP concentrations mediated by the calcium–calmodulin complex. The initial strong CBF enhancement in response to ACh is mainly governed by PKG and elevated calcium. The second phase of CBF enhancement induced by ACh, a stable moderately elevated CBF, is mainly regulated by PKA in a Ca²⁺-independent manner. Inhibition of either guanylate cyclase or of PKG partially attenuates the response to ACh of [Ca²⁺]_i, but completely abolishes the response of CBF. Inhibition of PKA moderately attenuates and significantly shortens the responses to ACh of both [Ca²⁺]_i and CBF. In addition, PKA facilitates the elevation in [Ca²⁺]_i and cGMP levels induced by ACh, whereas an unimpeded PKG activity is essential for CBF enhancement mediated by either Ca²⁺ or PKA.

Key Words: cilia • mucociliary tissue • phosphorylation • cholinergic • cyclic nucleotides

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