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First published online October 2, 2009
Journal of Experimental Biology 212, 3211-3220 (2009)
Published by The Company of Biologists 2009
doi: 10.1242/jeb.029108
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Atrial granular cells of the snail Achatina fulica release proteins into hemolymph after stimulation of the heart nerve

Sergej V. Shabelnikov*, Olga A. Bystrova, Vadim A. Ivanov, Boris A. Margulis and Marina Martynova

Institute of Cytology, Russian Academy of Sciences, Tikhoretsky Avenue 4, St Petersburg 194064, Russia

* Author for correspondence (buddasvami{at}gmail.com)

Accepted 7 July 2009

The atrium of the gastropod mollusc Achatina fulica receives rich innervation and contains numerous granular cells (GCs). We studied the atrial innervation and discovered that axon profiles typical in appearance of peptidergic neurons form close unspecialized membrane contacts with GCs. Then, we investigated, at both morphological and biochemical levels, the effect of electrical stimulation of the heart nerve on GCs of Achatina heart perfused in situ. The ultrastructural study demonstrated changes in granule morphology consistent with secretion. These events included alteration of granule content, intracellular granule fusion and formation of complex degranulation channels, within which the granule matrix solubilized. It was shown that electrical stimulation resulted in a significant increase of the total protein concentration in the perfusate. Furthermore, SDS-PAGE analysis of the perfusate revealed three new proteins with molecular masses of 16, 22, and 57 kDa. Affinity-purified polyclonal antibodies against the 16 kDa protein were obtained; the whole-mount immunofluorescence technique revealed the presence of this protein in the granules of atrial GCs. In GCs of the stimulated atrium, a progressive loss of their granular content was observed. The results suggest that the central nervous system can modulate the secretory activity of the atrial GCs through non-synaptic pathways.

Key words: axon, degranulation, molluscs, non-synaptic contacts, protein secretion, ultrastructure


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