Five ADNFLE mutations reduce the Ca2+ dependence of the mammalian alpha4beta2 acetylcholine response.

Abstract

Five nicotinic acetylcholine receptor (nAChR) mutations are currently linked to autosomal dominant nocturnal frontal lobe epilepsy (ADNFLE). The similarity of their clinical symptoms suggests that a common functional anomaly of the mutations underlies ADNFLE seizures. To identify this anomaly, we constructed rat orthologues (S252F, +L264, S256L, V262L, V262M) of the human ADNFLE mutations, expressed them in Xenopus oocytes with the appropriate wild-type (WT) subunit (alpha4 or beta2), and studied the Ca2+ dependence of their ACh responses. All the mutations significantly reduced 2 mM Ca2+-induced increases in the 30 microM ACh response (P < 0.05). Consistent with a dominant mode of inheritance, this reduction persisted in oocytes injected with a 1:1 mixture of mutant and WT cRNA. BAPTA injections showed that the reduction was not due to a decrease in the secondary activation of Ca2+-activated Cl- currents. The S256L mutation also abolished 2 mM Ba2+ potentiation of the ACh response. The S256L, V262L and V262M mutations had complex effects on the ACh concentration-response relationship but all three mutations shifted the concentration-response relationship to the left at [ACh] >= 30 microM. Co-expression of the V262M mutation with a mutation (E180Q) that abolished Ca2+ potentiation resulted in 2 mM Ca2+ block, rather than potentiation, of the 30 microM ACh response, suggesting that the ADNFLE mutations reduce Ca2+ potentiation by enhancing Ca2+ block of the alpha4beta2 nAChR. Ca2+ modulation may prevent presynaptic alpha4beta2 nAChRs from overstimulating glutamate release at central excitatory synapses during bouts of synchronous, repetitive activity. Reducing the Ca2+ dependence of the ACh response could trigger seizures by increasing alpha4beta2-mediated glutamate release during such bouts.

• Authors: Nivalda Rodrigues-Pinguet, Li Jia, Maureen Li, Antonio Figl, Alwin Klaassen, Anthony Truong, Henry A Lester, Bruce N Cohen
• Journal: The Journal of physiology (J Physiol) Vol. 550 Issue Pt 1 Pg. 11-26 (Jul 01 2003) ISSN: 0022-3751 [Print] England
• PMID: 12754307 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Bridged Bicyclo Compounds, Heterocyclic
• Chloride Channels
• Nicotinic Agonists
• Pyridines
• Receptors, Nicotinic
• nicotinic receptor alpha4beta2
• Barium
• epibatidine
• Acetylcholine
• Calcium

Topics

• Acetylcholine (pharmacology)
• Amino Acid Sequence (genetics)
• Animals
• Artifacts
• Barium (pharmacology)
• Bridged Bicyclo Compounds, Heterocyclic (metabolism)
• Calcium (pharmacology, physiology)
• Cell Membrane (metabolism)
• Chloride Channels (physiology)
• Circadian Rhythm (genetics)
• Electric Conductivity
• Epilepsy, Frontal Lobe (genetics)
• Genes, Dominant
• Homeostasis
• Molecular Sequence Data
• Mutation (genetics, physiology)
• Nicotinic Agonists (metabolism)
• Oocytes
• Pyridines (metabolism)
• Rats
• Reaction Time
• Receptors, Nicotinic (drug effects, genetics, metabolism)
• Xenopus laevis