title: Analysis of Ca(2+) uptake into the smooth endoplasmic reticulum of permeabilised sternal epithelial cells during the moulting cycle of the terrestrial isopod Porcellio scaber.
abstract: In terrestrial isopods, large amounts of Ca(2+) are transported across anterior sternal epithelial cells during moult-related deposition and resorption of CaCO(3) deposits. Because of its toxicity and function as a second messenger, resting cytosolic Ca(2+) levels must be maintained below critical concentrations during epithelial Ca(2+) transport, raising the possibility that organelles play a role during Ca(2+) transit. We therefore studied the uptake of Ca(2+) into Ca(2+)-sequestering organelles by monitoring the formation of birefringent calcium oxalate crystals in permeabilised anterior and posterior sternal epithelium cells of Porcellio scaber during Ca(2+)-transporting and non-transporting stages of the moulting cycle using polarised-light microscopy. The results indicate ATP-dependent uptake of Ca(2+) into organelles. Half-maximal crystal growth at a Ca(2+) activity, a(Ca), of 0.4 micromol l(-1) and blockade by cyclopiazonic acid suggest Ca(2+) uptake into the smooth endoplasmic reticulum by the smooth endoplasmic reticulum Ca(2+)-ATPase. Analytical electron microscopical techniques support this interpretation by revealing the accumulation of Ca(2+)-containing crystals in smooth membranous intracellular compartments. A comparison of different moulting stages demonstrated a virtual lack of crystal formation in the early premoult stage and a significant fivefold increase between mid premoult and the Ca(2+)-transporting stages of late premoult and intramoult. These results suggest a contribution of the smooth endoplasmic reticulum as a transient Ca(2+) store during intracellular Ca(2+) transit.
authors.affiliation: Zentrale Einrichtung Elektronenmikroskopie, Universität Ulm, 89069 Ulm, Germany.
authors.forename: Monica
authors.initials: M
authors.lastname: Hagedorn
authors.forename: Andreas
authors.initials: A
authors.lastname: Ziegler
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002129:Calcium Oxalate / Q000378:metabolism; D018871:Endoplasmic Reticulum, Smooth / Q000378:metabolism*; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism*; D004848:Epithelium / Q000378:metabolism / Q000648:ultrastructure; D017136:Ion Transport; D033321:Isopoda / Q000378:metabolism*; D018978:Molting / Q000502:physiology*; D013057:Spectrum Analysis / Q000379:methods
chemical_list: D002129:Calcium Oxalate; D002118:Calcium