Cellular prion protein is implicated in the regulation of local Ca2+ movements in cerebellar granule neurons.

The cellular prion protein (PrP(C)) is a cell-surface glycoprotein mainly expressed in the CNS. The structural conversion of PrP(C) generates the prion, the infectious agent causing transmissible spongiform encephalopathies, which are rare and fatal diseases affecting animals and humans. Despite decades of intensive research, the mechanism of prion-associated neurodegeneration and the physiologic role of PrP(C) are still obscure. Recent evidence, however, supports the hypothesis that PrP(C) may be involved in the control of Ca(2+) homeostasis. Given the universal significance of Ca(2+) as an intracellular messenger for both the life and death of cells, this possibility may help explain the complex, often controversial, dataset accumulated on PrP(C) physiology, and the events leading to prion-associated neuronal demise. In this study, we have compared local Ca(2+) movements in cerebellar granule neurons (CGN) derived from wild-type (WT), or PrP-knockout (KO), mice, by means of the Ca(2+)-sensitive photo-probe, aequorin, genetically targeted to specific intracellular domains and delivered to CGN by lentiviral vectors. The use of an aequorin that localizes to the cytosolic domains proximal to the plasma membrane has allowed us to demonstrate that there was a dramatic increase of store-operated Ca(2+) entry in PrP-KO CGN compared to WT neurons. Notably, this phenotype was rescued upon restoring PrP(C) expression. The Ca(2+)-phenotype of PrP-KO neurons can in part be explained by the lower expression of two major Ca(2+)-extruding proteins, namely the plasma membrane and the sarco-endoplasmic reticulum Ca(2+)-ATPases. The lower sarco-endoplasmic reticulum Ca(2+)-ATPase content may also contribute to explain why PrP-KO CGN accumulated less Ca(2+) in the endoplasmic reticulum than the WT counterpart.