[Beta]-1, 3-glucan modulates PKC signalling in Lymnaea stagnalis defence cells: a role for PKC in H[sub]2O[sub]2 production and downstream ERK activation

Lacchini, Audrey H., Davies, Angela J., Mackintosh, David and Walker, Anthony J. (2006) [Beta]-1, 3-glucan modulates PKC signalling in Lymnaea stagnalis defence cells: a role for PKC in H[sub]2O[sub]2 production and downstream ERK activation. The Journal of Experimental Biology (JEB), 209(24), pp. 4829-4840. ISSN (print) 0022-0949

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Haemocytes from the gastropod snail Lymnaea stagnalis (Linnaeus) were used as a model to characterize protein kinase C (PKC) signalling events in molluscan defence cells. Challenge of freshly collected haemocytes with the beta-1, 3-glucan laminarin resulted in a transient increase in the phosphorylation of haemocyte PKC, with maximal phosphorylation (represented by a 3.5-fold increase) occurring at 10 min; this effect was blocked by the PKC inhibitor, GF109203X. Moreover, extracellular signal-regulated kinase (ERK) was found to be a downstream target of molluscan PKC, operating via a MAPK/ERK kinase (MEK)-dependent mechanism. Pharmacological inhibition of PKC phosphorylation by U-73122 and ET-18-OCH(3) suggested that laminarin-dependent PKC signalling was modulated via phospholipase C (PLC); however, a role for phosphatidylinositol-3-kinase (PI-3-K) is unlikely since the PI-3-K inhibitor LY294002 was without effect. Generation of H(2)O(2) by haemocytes in response to laminarin was also investigated. H(2)O(2) output increased in a dose- and time-dependent manner, with 10 mg ml(-1) laminarin eliciting a 9.5-fold increase in H(2)O(2) production after 30 min. H(2)O(2) production was significantly attenuated by the PKC inhibitors, GF109203X and Gö 6976, and by the NADPH-oxidase inhibitor, apocynin. In conclusion, these data further our understanding of PKC signalling events in molluscan haemocytes and for the first time define a role for PKC in H(2)O(2) production by these defence cells. Given that H(2)O(2) is an important anti-pathogen molecule, and that haemocytes play a crucial role in the elimination of invading organisms, PKC signalling in these cells is likely to be crucial to the molluscan innate defence response.

Item Type: Article
Uncontrolled Keywords: haemocyte, mollusc defence, PKC, ERK, hydrogen peroxide, reactive oxygen species
Research Area: Allied health professions and studies
Faculty, School or Research Centre: Faculty of Science (until 2011) > School of Life Sciences
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Depositing User: Sara Burnett
Date Deposited: 25 Jun 2007
Last Modified: 04 May 2011 08:26
DOI: https://doi.org/10.1242/jeb.02561
URI: http://eprints.kingston.ac.uk/id/eprint/1487

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