Reduction in CHT1-mediated choline uptake in primary neurons from presenilin-1 M146V mutant knock-in mice.
The memory loss in Alzheimer's disease (AD) has been linked to cholinergic hypoactivity. Mutations in presenilin-1 (PS-1) may regulate cholinergic signaling, although their precise roles in cholinergic neurotransmission in AD are unsettled. Neuronal uptake of choline via the high affinity choline transporter (CHT1) is essential for cholinergic neurotransmission. CHT1 is a Na+-dependent, hemicholinium-3 (HC-3)-sensitive choline transporter. Although cholinergic neurons in the nucleus basalis of Meynert are a major source of cholinergic projections for the cerebral cortex, it is unclear whether cortical neurons exhibit intrinsic CHT1 activity that is altered in AD. We now report that primary cortical neurons express intrinsic and biologically active CHT1, and that, in these neurons, CHT1-mediated choline uptake activity is significantly reduced in PS-1 M146V mutant knock-in mice. Further kinetic studies using HC-3 binding and cell surface biotinylation assays showed that the PS-1 mutation inhibits CHT1 mediated choline uptake by reducing the ligand binding affinity of CHT1 without significantly altering levels of CHT1 expression in the plasma membrane. Since human neocortex has recently been shown to possess intrinsic cholinergic innervation, our results indicate that alterations in CHT1-mediated high affinity choline uptake in cortical neurons may contribute to Alzheimer's dementia.
Payette
Daniel J
DJ
Department of Physiology, the University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA.
Xie
Jun
J
Guo
Qing
Q
D000704:Analysis of Variance; D000818:Animals; D002540:Cerebral Cortex / Q000166:cytology; D002794:Choline / Q000378:metabolism* / Q000493:pharmacokinetics; D002795:Choline O-Acetyltransferase / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D006426:Hemicholinium 3 / Q000494:pharmacology; D026901:Membrane Transport Proteins / Q000378:metabolism*; D008715:Methionine / Q000235:genetics*; D051379:Mice; D008822:Mice, Transgenic; D009154:Mutation; D009474:Neurons / Q000187:drug effects / Q000378:metabolism*; D014179:Neurotransmitter Uptake Inhibitors / Q000494:pharmacology; D053764:Presenilin-1 / Q000235:genetics*; D011485:Protein Binding / Q000187:drug effects; D013997:Time Factors; D014633:Valine / Q000235:genetics*
C488255:CHT1 protein, mouse; D026901:Membrane Transport Proteins; D014179:Neurotransmitter Uptake Inhibitors; D053764:Presenilin-1; D006426:Hemicholinium 3; D008715:Methionine; D002795:Choline O-Acetyltransferase; D014633:Valine; D002794:Choline