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Am J Physiol Regul Integr Comp Physiol (April 3, 2003). doi:10.1152/ajpregu.00634.2002
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Submitted on October 11, 2002
Accepted on March 25, 2003

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

Shandra A Doran1, Cam Ha Tran1, Cagla Eskicioglu1, Tev Stachniak2, Kee-Chan Ahn1, and Jeffrey I Goldberg3*

1 Biological Sciences, University of Alberta, Edmonton, Alberta, Canada
2 Center for Neuroscience, University of Alberta, Edmonton, Alberta, Canada
3 Biological Sciences, University of Alberta, Edmonton, Alberta, Canada; Center for Neuroscience, University of Alberta, Edmonton, Alberta, Canada

* To whom correspondence should be addressed. E-mail: jeff.goldberg{at}ualberta.ca.

Embryos of Helisoma trivolvis exhibit cilia-driven rotation within the egg capsule during development. In this study we examined if nitric oxide (NO) is a physiological regulator of ciliary beating in cultured ciliary cells. The NO donor S-Nitroso-N-acetylpenicillamine (SNAP; 1-1000 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. While 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 analogue 8-Br-cGMP (0.2 and 2 mM) increased CBF and the soluble guanylate cyclase inhibitor LY 83583 (10 µM) blocked the cilio-excitatory effects of SNAP and serotonin. These data suggest that NO has a constitutive cilio-excitatory 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 cilio-excitatory action of serotonin.




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