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In vitro cultured cerebellar granule cells (cgc) undergo apoptotic death when the depolarising concentration of KCl 25 mM is adjusted to 5 mM. We investigated whether the apoptotic message can be transferred from a group of neurons to neighbouring but separated sister cultures by resorting to two different culture systems, one based on the use of concentric dishes, the other employing the transwells. The extent of transferred death was 38.6% in the two dishes system and 31.3% in the transwell system. The transfer of such death-inducing signal(s), accompanied by chromatin condensation, was inhibited by actinomicin D. Preliminary experiments aimed at identifying such apoptosis-inducing signal(s) suggest the involvement of beta-amyloid fragment(s).

We have recently shown that, in rat cerebellar granule cells, apoptosis triggered by KCl deprivation is associated with an amyloidogenic shift in the processing of amyloid precursor protein (APP) resulting in an increase of amyloid beta-protein (A beta) secretion. To further investigate this issue we studied the relationship between secretion of APP metabolites (A beta, APPs) and neuronal degeneration. We postulated that the endogenous products of the APP metabolism may modulate neuronal survival by an autocrine loop. Treatment of cerebellar granule cells with various antibodies raised against different epitopes of APPs and A beta oppositely modulates low potassium apoptotic cell death. Antibodies specific for the N-terminal of A beta (4G8, 6E10, R3659) increased neuronal survival by 30% over controls. On the contrary, treatment of cultures undergoing apoptosis with the monoclonal antibody 22C11 directed against the APP N-terminus reduced neuronal survival by 53%, suggesting that endogenous alpha-APPs contribute to neuronal survival. Moreover low KCl culture medium, conditioned by cerebellar granule cells, attenuated the apoptotic process. This anti-apoptotic effect was abolished by removal of APPs from the conditioned medium. Western blotting of APPs removed from the conditioned medium confirmed the presence of alpha-APPs. These data indicate that APP cleavage products oppositely modulate neuronal survival through an autocrine loop and further strengthen an Alzheimer's disease pathogenetic scheme based on altered metabolism of APP.

Experimental evidence of the preferential uptake of aluminium by GABAergic neurones and glial cells was provided by synchrotron spectromicroscopy studies. We observed rat cerebellar cultures enriched for GABAergic neurones or glial cells exposed to aluminium ions, detecting the presence and identifying the chemical status of aluminium on cell structures.

A systematic microchemical analysis of unstained and uncoated neurone cultures was performed with synchrotron radiation photoemission spectromicroscopy after exposure to an aluminium solution. Clear evidence was found for localized aluminium uptake in a few cells. Their possible identification based on morphology is discussed.

We present the first comparative study of the uptake of metal ions by neurons, performed for Zn, Cr, Co, Mo, Al, Ni, Mn and Cd. The study reveals substantial differences in the uptake of different metals, under similar exposure procedures. In particular, we found very large uptakes for aluminium and molybdenum. We also found significant effects of excitatory substances, in particular kainate, as stimulants of uptake of some of the metals.