The non-reducing disaccharide trehalose is biosynthesized in several species but not in vertebrates. However, trehalase, the enzyme required for its cleavage, has been observed in different mammalian organs. Even in humans, trehalase was detected in the gastrointestinal tract and the kidney. Trehalase is an intrinsic glycoprotein of the small intestine and kidney that transports trehalose and hydrolyses it to two glucose molecules. To our knowledge, no information is available about the in vivo distribution and localization of trehalase in the mammalian brain. Here, we report the occurrence and distribution of trehalase in vivo in the mouse brain using Western blotting and immunohistochemical techniques. Using an antibody against trehalase, we demonstrated that the enzyme showed a band with a molecular mass of approx. 70 kDa in the hippocampus, cerebral cortex, cerebellum and olfactory bulbs. Strong trehalase immunoreactivity was found in the perikarya and dendrites of neurons located in the hippocampus, cerebral cortex, Purkinje cells and mitral cells. Interestingly, Purkinje cells of the cerebellum showed higher immunoreactivity than neurons in the hippocampus and cerebral cortex. The distribution of trehalase appeared to be mainly related to neurons and was not detected in astrocytes. Independent of the presence of trehalose in neurons, the trehalase levels in neurons should have physiological significance. Investigating whether the interactions between trehalose and trehalase act on brain energy metabolism or have other not-yet-identified effects would also be interesting.

Accumulating evidence indicates that loss of physiologic amyloid precursor protein (APP) function leads to reduced neuronal plasticity, diminished synaptic signaling and enhanced susceptibility of neurons to cellular stress during brain aging. Here we investigated the neuroprotective function of the soluble APP ectodomain sAPPα (soluble APPα), which is generated by cleavage of APP by α-secretase along the non-amyloidogenic pathway. Recombinant sAPPα protected primary hippocampal neurons and SH-SY5Y neuroblastoma cells from cell death induced by trophic factor deprivation. We show that this protective effect is abrogated in neurons from APP-knockout animals and APP-depleted SH-SY5Y cells, but not in APP-like protein 1- and 2- (APLP1 and APLP2) depleted cells, indicating that expression of membrane-bound holo-APP is required for sAPPα-dependent neuroprotection. Trophic factor deprivation diminished the activity of the Akt survival pathway. Strikingly, both recombinant sAPPα and the APP-E1 domain were able to stimulate Akt activity in wild-type (wt) fibroblasts, SH-SY5Y cells and neurons, but failed to rescue in APP-deficient neurons or fibroblasts. The ADAM10 (a disintegrin and metalloproteinase domain-containing protein 10) inhibitor GI254023X exacerbated neuron death in organotypic (hippocampal) slice cultures of wt mice subjected to trophic factor and glucose deprivation. This cell death-enhancing effect of GI254023X could be completely rescued by applying exogenous sAPPα. Interestingly, sAPPα-dependent Akt induction was unaffected in neurons of APP-ΔCT15 mice that lack the C-terminal YENPTY motif of the APP intracellular region. In contrast, sAPPα-dependent rescue of Akt activation was completely abolished in APP mutant cells lacking the G-protein interaction motif located in the APP C-terminus and by blocking G-protein-dependent signaling with pertussis toxin. Collectively, our data provide new mechanistic insights into the physiologic role of APP in antagonizing neurotoxic stress: they suggest that cell surface APP mediates sAPPα-induced neuroprotection via G-protein-coupled activation of the Akt pathway.