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Calcium signalling is little understood in filamentous fungi largely because easy and routine methods for calcium measurement in living hyphae have previously been unavailable. We have developed the recombinant aequorin method for this purpose. High levels of aequorin expression were obtained in Neurospora crassa, Aspergillus niger and Aspergillus awamori by codon optimization of the aequorin gene. Three external stimuli (mechanical perturbation, hypo-osmotic shock and high external calcium) were found transiently to increase [Ca2+](c). Each of the calcium signatures associated with these physico-chemical treatments was unique, suggesting the involvement of three distinct calcium-mediated signal transduction pathways. The fungal calcium channel blocker KP4 inhibited the [Ca2+](c) responses to hypo-osmotic shock and high external calcium, but not to mechanical perturbation. The divalent cation chelator BAPTA inhibited [Ca2+](c) responses to mechanical perturbation and hypo-osmotic shock. The calcium agonists A23187 and cyclopiazonic acid increased [Ca2+](c) levels.
Author(s): Nelson G, Kozlova-Zwinderman O, Collis AJ, Knight MR, Fincham JRS, Stanger CP, Renwick A, Hessing JGM, Punt PJ, van den Hondel CAMJJ, Read ND
Publication type: Article
Publication status: Published
Journal: Molecular Microbiology
Print publication date: 01/06/2004
ISSN (print): 0950-382X
ISSN (electronic): 1365-2958
Publisher: Blackwell Publishing Ltd
Cited References Count:86
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