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J Physiol Volume 515, Number 2, 511-522, March 1, 1999
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The Journal of Physiology (1999), 515.2, pp. 511-522
© Copyright 1999 The Physiological Society

Involvement of protein kinase C in 5-HT-stimulated ciliary activity in Helisoma trivolvis embryos

Kimberley J. Christopher, Kevin G. Young, John P. Chang and Jeffrey I. Goldberg

Department of Biological Sciences, CW-405 Biological Sciences Building, University of Alberta, Edmonton, Alberta, Canada T6G 2E9


During development, embryos of the pulmonate gastropod, Helisoma trivolvis, undergo a rotation behaviour due to the co-ordinated beating of three bands of ciliated epithelial cells. This behaviour is in part mediated by the neurotransmitter serotonin (5-HT) released from a pair of identified embryonic neurons. Using time-lapse videomicroscopy to measure ciliary beat frequency (CBF) in response to pharmacological manipulations, we determined whether protein kinase C (PKC) is involved in mediating 5-HT-stimulated ciliary beating.


Diacylglycerol (DAG) analogues sn-1,2-dioctanoyl glycerol (DiC8; 100 µM) and 1-oleoyl-2-acetyl-sn-glycerol (OAG; 100 µM), partially mimicked the 5-HT-induced increase in CBF. In contrast, application of OAG in the absence of extracellular Ca2+ did not result in an increase in CBF.


5-HT-stimulated CBF was effectively blocked by PKC inhibitors bisindolylmaleimide (10 and 100 nM) and calphostin C (10 nM). In addition, bisindolylmaleimide (100 nM) inhibited DiC8-induced increases in CBF. At a higher concentration (200 nM), bisindolylmaleimide did not significantly reduce 5-HT-stimulated cilio-excitation.


Two different phorbol esters, phorbol 12-myristate 13-acetate (TPA; 0·1, 10 or 1000 nM) and phorbol 12beta, 13alpha-dibenzoate (PDBn; 10 µM) did not alter basal CBF. TPA (1 µM) did not alter 5-HT-stimulated CBF. Likewise, the synthetic form of phosphatidylserine,
N-(6-phenylhexyl)-5-chloro-1-naphthalenesulphonamide (SC-9; 10 µM), did not increase CBF, whereas a strong increase in CBF was observed upon exposure to 5-HT.


The results suggest that a DAG-dependent, phorbol ester-insensitive isoform of PKC mediates 5-HT-stimulated CBF in ciliated epithelial cells from embryos of Helisoma trivolvis.


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