Reduced ACh release at neuromuscular synapses of heterozygous leaner Ca v 2.1‐mutant mice

Episodic ataxia type 2 (EA2) is an autosomal dominantly inherited neurological disorder. Patients have CACNA1A gene mutations resulting in truncation or single amino acid changes in the pore‐forming subunit of Ca v 2.1 (P/Q‐type) Ca 2+ channels. These neuronal channels mediate synaptic neurotransmitter release. EA2 symptoms are thought to result from disturbed neurotransmission at cerebellar and neuromuscular synapses, caused by loss‐of‐function of Ca v 2.1 channels. Heterozygous leaner ( Ln /wt) mice, carrying a Cacna1a truncation mutation, as well as heterozygous Ca v 2.1 null ‐mutant (KO/wt) mice may model synaptic aspects of EA2. We studied Ca v 2.1‐mediated acetylcholine (ACh) release at their neuromuscular junctions (NMJs) ex vivo. KO/wt mice did not show any ACh release abnormalities, not even at older age. However, Ln /wt mice had ∼25% reduced spontaneous uniquantal ACh release and ∼10% reduced nerve‐stimulation evoked release, compared with wild‐type. EA2 is treated with acetazolamide (AZA), but the pharmacotherapeutic mechanism is unknown. We tested the possibility of a direct influence on (mutant) presynaptic Ca v 2.1 channel function by studying the acute effect of 50 μM AZA on ACh release at ex vivo NMJs of wild‐type, KO/wt, and Ln /wt mice. No changes were found in any of the release parameters. Our results indicate that Ln ‐mutated Ca v 2.1 channels at Ln /wt NMJs are either normally inserted in the presynaptic membrane but have reduced function, or that they inhibit wild‐type channels by hampering their expression, trafficking, membrane insertion and/or function. In this respect Ln /wt NMJs may model EA2 synapses. Furthermore, AZA does not exert an acute, direct influence on the function of presynaptic (mutant) Ca v 2.1 channels. Synapse 62:337–344, 2008. © 2008 Wiley‐Liss, Inc.