Disruption of synaptosomal calcium channel function by Lambert-Eaton myasthenic immunoglobulin is serum-dependent.
An autoantibody to nerve terminal Ca2+ channels has been suggested to mediate the pathogenesis of the neuromuscular disorder Lambert-Eaton Myasthenic Syndrome (LEMS). We demonstrated previously that in the presence of control human serum, immunoglobulins isolated from a patient with LEMS reduced flux of Ca2+ into isolated nerve terminals during depolarization. The objective of the present study was to determine the role of serum in reducing uptake of 45Ca2+ into rat brain synaptosomes by LEMS IgG. Depolarization-dependent uptake of 45Ca2+ through voltage-gated Ca2+ channels was determined using synaptosomes incubated with control (disease-free) and LEMS IgG with or without control human serum. In the absence of human serum, LEMS IgG did not reduce uptake of 45Ca2+ into synaptosomes. However, in the presence of control human serum (10% of total incubation volume), 45Ca2+ uptake was reduced significantly by LEMS IgG (2 and 4 mg/ml), but not by IgG from disease-free patients or by 10% (v/v) control human serum alone. This concentration of serum was found to be optimal; higher concentrations produced significant reductions in Ca2+ uptake, whereas at lower concentrations the serum/IgG combination was ineffective. The depressant effect of high concentrations of serum alone on 45Ca2+ uptake was mimicked by equal concentrations of bovine serum albumin suggesting that deficits in 45Ca2+ uptake produced by high concentrations of serum were due to increased protein binding of the radiolabel. Heat-inactivating the serum abolished its ability to interact with the LEMS immunoglobulins to depress 45Ca2+ uptake. This suggested a role for complement in this effect.(ABSTRACT TRUNCATED AT 250 WORDS)
Hewett
S J
SJ
Department of Pharmacology/Toxicology, Michigan State University, East Lansing 48824-1317.
Atchison
W D
WD
D000818:Animals; D001790:Blood Physiological Phenomena; D015220:Calcium Channels / Q000276:immunology*; D002132:Calcium Radioisotopes; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology*; D015624:Lambert-Eaton Myasthenic Syndrome / Q000276:immunology* / Q000378:metabolism; D008297:Male; D018377:Neurotransmitter Agents / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013574:Synaptosomes / Q000276:immunology*
D015220:Calcium Channels; D002132:Calcium Radioisotopes; D007074:Immunoglobulin G; D018377:Neurotransmitter Agents