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DEVELOPMENT AND TISSUE PLASTICITY

Constitutive and permissive roles of nitric oxide activity in embryonic ciliary cells

Department of Biological Sciences, University of Alberta, Edmonton, Alberta, Canada T6G 2E9

Submitted 11 October 2002 ; accepted in final form 25 March 2003

Embryos of Helisoma trivolvis exhibit cilia-driven rotation within the egg capsule during development. In this study we examined whether nitric oxide (NO) is a physiological regulator of ciliary beating in cultured ciliary cells. The NO donor S-nitroso-N-acetylpenicillamine (SNAP; 1–1,000 µM) produced a dose-dependent increase in ciliary beat frequency (CBF). In contrast, the nitric oxide synthase (NOS) inhibitor 7-nitroindazole (10 and 100 µM) inhibited the basal CBF and blocked the stimulatory effects of serotonin (100 µM). NO production in response to serotonin was investigated with 4,5-diaminofluorescein diacetate imaging. Although SNAP (100 µM) produced a rise in NO levels in all cells, only 22% of cells responded to serotonin with a moderate increase. The cGMP analog 8-bromo-cGMP (8-Br-cGMP; 0.2 and 2 mM) increased CBF, and the soluble guanylate cyclase inhibitor LY-83583 (10 µM) blocked the cilioexcitatory effects of SNAP and serotonin. These data suggest that NO has a constitutive cilioexcitatory effect in Helisoma embryos and that the stimulatory effects of serotonin and NO work through a cGMP pathway. It appears that in Helisoma cilia, NO activity is necessary, but not sufficient, to fully mediate the cilioexcitatory action of serotonin.

Helisoma; mollusk; ciliary beat frequency; 4,5-diaminofluorescein; serotonin

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				S. A. Doran, R. Koss, C. H. Tran, K. J. Christopher, W. J. Gallin, and J. I. Goldberg Effect of serotonin on ciliary beating and intracellular calcium concentration in identified populations of embryonic ciliary cells J. Exp. Biol., March 15, 2004; 207(8): 1415 - 1429. [Abstract] [Full Text] [PDF]