, Volume 21, Issue 6, pp 305-313

Susceptibility of cerebellar granule neurons from GM2/GD2 synthase-null mice to apoptosis induced by glutamate excitotoxicity and elevated KCl: Rescue by GM1 and LIGA20

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Abstract

Our previous study showed an impaired regulation of Ca2+ homeostasis in cultured cerebellar granule neurons (CGN) from neonatal mice lacking GM2, GD2 and all gangliotetraose gangliosides, due to disruption of the GM2/GD2 synthase (GalNAc-T) gene. In the presence of depolarizing concentration (55 mM) K+, these cells showed persistent elevation of intracellular Ca2+ ([Ca2+] i ) leading to apoptosis and cell destruction. This was in contrast to CGN from normal littermates whose survival was enhanced by high K+. In this study we demonstrate that glutamate has the same effect as K+ on CGN from these ganglioside-deficient knockout (KO) mice and that apoptosis in both cases is averted by exogenous GM1. Even more effective rescue was obtained with LIGA20, a semi-synthetic derivative of GM1. LC50 of glutamate in the KO cells was 3.1 μM, compared to 46 μM in normal CGN. [Ca2+] i measurement with fura-2 revealed no difference in glutamate-stimulated Ca2+ influx between the 2 cell types. However, reduction of [Ca2+] i following application of Mg2+ was significantly impaired in the mutant CGN. The rescuing effects of exogenous GM1 and LIGA20 corresponded to their ability to restore Ca2+ homeostasis. The greater potency of LIGA20 is attributed to its greater membrane permeability with resultant ability to insert into both plasma and nuclear membranes at low concentration (≤1μM); GM1 at the same concentration was incorporated only into the plasma membrane and required much higher concentration to influence Ca2+ homeostasis and CGN viability. Published in 2004.

This revised version was published online in July 2006 with corrections to the Cover Date.