Analysis of Ca2+ uptake into the smooth endoplasmic reticulum of permeabilised sternal epithelial cells during the moulting cycle of the terrestrial isopod Porcellio scaber

摘要

In terrestrial isopods, large amounts of Ca2+ are transported across anterior sternal epithelial cells during moult-related deposition and resorption of CaCO3 deposits. Because of its toxicity and function as a second messenger, resting cytosolic Ca2+ levels must be maintained below critical concentrations during epithelial Ca2+ transport, raising the possibility that organelles play a role during Ca2+ transit. We therefore studied the uptake of Ca2+ into Ca2+-sequestering organelles by monitoring the formation of birefringent calcium oxalate crystals in permeabilised anterior and posterior sternal epithelium cells of Porcellio scaber during Ca2+-transporting and non-transporting stages of the moulting cycle using polarised-light microscopy. The results indicate ATP-dependent uptake of Ca2+ into organelles. Half-maximal crystal growth at a Ca2+ activity, a(Ca), of 0.4 mumol l(-1) and blockade by cyclopiazonic acid suggest Ca2+ uptake into the smooth endoplasmic reticulum by the smooth endoplasmic reticulum Ca2+-ATPase. Analytical electron microscopical techniques support this interpretation by revealing the accumulation of Ca2+-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 Ca2+-transporting stages of late premoult and intramoult. These results suggest a contribution of the smooth endoplasmic reticulum as a transient Ca2+ store during intracellular Ca2+ transit.