Amyotrophic Lateral Sclerosis Immunoglobulins Increase Intracellular Calcium in a Motoneuron Cell Line

A hybrid motoneuron cell line (VSC4.1) was used as a model system to study the relationship between alterations in intracellular calcium and subsequent cell death induced by immunoglobulin fractions purified from sera of patients with ALS. Using fluo-3 fluorescence imaging, immunoglobulins from 8 of 10 patients with ALS were found to induce transient increases in intracellular calcium (Ca²⁺]_i) in differentiated VSC4.1 cells. These transient Ca²⁺]_iincreases required extracellular calcium entry through voltage-gated calcium channels sensitive to synthetic FTX and to high concentrations (>1 μM) of ω-agatoxin IVa. The incidence of transient Ca²⁺]_iincreases induced by ALS immunoglobulins correlated with the extent of cytotoxicity induced by the same ALS immunoglobulins in parallel cultures of VSC4.1 cells. Furthermore, manipulations which blocked transient Ca²⁺]_iincreases (addition of synthetic FTX or ω-agatoxin IVa) also inhibited the cytotoxic effects of ALS immunoglobulins. No transient calcium increases were observed in VSC4.1 cells following addition of immunoglobulins from 7 neurologic disease control patients. However, transient Ca²⁺]_iincreases were observed following addition of immunoglobulins from 4 of 5 patients with myasthenia gravis (MG). The Ca²⁺]_ichanges induced by MG immunoglobulins were not blocked by s-FTX, suggesting that they result from a different mechanism than those induced by ALS immunoglobulins. These results suggest that immunoglobulins from patients with ALS can induce transient increases in intracellular calcium in a motoneuron cell line, which may represent early events in the cascade of processes leading to injury and death of susceptible cells.