Mineral Fabrication and Golgi Apparatus Activity in <i>Spirostomum ambiguum</i>: A Primordial Paradigm of the Stressed Bone Cell? — Oak Academic Publishing
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Mineral Fabrication and Golgi Apparatus Activity in <i>Spirostomum ambiguum</i>: A Primordial Paradigm of the Stressed Bone Cell?
Institute of Membrane and Systems Biology, Faculty of Biological Sciences, University of Leeds, Leeds, UK
,
Institute of Membrane and Systems Biology, Faculty of Biological Sciences, University of Leeds, Leeds, UK
,
Institute of Membrane and Systems Biology, Faculty of Biological Sciences, University of Leeds, Leeds, UK
1 Institute of Membrane and Systems Biology, Faculty of Biological Sciences, University of Leeds, Leeds, UK
2 Institute of Membrane and Systems Biology, Faculty of Biological Sciences, University of Leeds, Leeds, UK
3 Institute of Membrane and Systems Biology, Faculty of Biological Sciences, University of Leeds, Leeds, UK
The histological basis for acute osteocyte mechanosensitivity remains uncertain. A novel bone cell model of mechanotransduction and inorganic trafficking may be the powerful, silt-burrowing protozoan Spirostomum ambiguum which when being physically challenged fabricates within vesicles populations of bone-like calcium phosphate microspheres, about 1 μm in diameter. These not only attribute considerable compression-resilience but also resemble the Golgi-directed mineral assemblies we recently reported in osteocytes. Advantageously, calcification in the protozoan (confirmed by ultramicroscopy with EDX elemental microanalysis) enabled Golgi comparison under overt, natural phases of both high ( i.e . silt-tunnelling) and low ( i.e free-swimming) stress. Established hard-tissue microscopy techniques previously positive in bone cells included quantitative fluorescent tetracycline labelling for bone salt together with the same metazoan Golgi body marker (Green Fluorescent Protein-tagged mannosidase II construct). Organellar modulation was monitored by transfection of live organisms in situ (some post-stained with red nuclear fluorochrome TOPRO-3). Results showed that GFP-tagged Golgi fluorescence increased from swimmers (mean 74.5 ± SD 6.7 AU) to burrowers (mean 104.6 ± SD 2.7; p < 0.0001) synchronous with juxtanuclear tetracycline-labelled mineral fluorescence (swimmers, mean 89.7 ± SD 3.3 AU; burrowers, mean 138.0 ± SD 4.0; p < 0.0001). Intracellular dense microspheres, single or bridged, were harvested as pellets rich in Ca, P (Ca:P 0.98) and Si, their polarised alignment moving from transaxial in swimmers to axial in burrowers. It was concluded that Golgi-directed mineral fabrication in the large, accessible, silt-enclosed ciliate resembles that in the smaller, less-accessible bone cell and may be a conserved early mechanobiological intracellular development predicating force translation into compression-resistant mineral fabrication in loaded segments of the osteocyte syncitium.
KeywordsGolgi-Directed CalcificationMechanosensing ProtozoanOsteocyte ModelTetracycline Fluorochrome for Bone MineralGFP Fluorochrome for Golgi Apparatus
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