title: Individual aggregates of amyloid beta induce temporary calcium influx through the cell membrane of neuronal cells.
abstract: Local delivery of amyloid beta oligomers from the tip of a nanopipette, controlled over the cell surface, has been used to deliver physiological picomolar oligomer concentrations to primary astrocytes or neurons. Calcium influx was observed when as few as 2000 oligomers were delivered to the cell surface. When the dosing of oligomers was stopped the intracellular calcium returned to basal levels or below. Calcium influx was prevented by the presence in the pipette of the extracellular chaperone clusterin, which is known to selectively bind oligomers, and by the presence a specific nanobody to amyloid beta. These data are consistent with individual oligomers larger than trimers inducing calcium entry as they cross the cell membrane, a result supported by imaging experiments in bilayers, and suggest that the initial molecular event that leads to neuronal damage does not involve any cellular receptors, in contrast to work performed at much higher oligomer concentrations.
authors.affiliation: Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge, UK.
authors.forename: Anna
authors.initials: A
authors.lastname: Drews
authors.affiliation: Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge, UK.
authors.forename: Jennie
authors.initials: J
authors.lastname: Flint
authors.affiliation: Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge, UK.
authors.forename: Nadia
authors.initials: N
authors.lastname: Shivji
authors.affiliation: Department of Chemistry, Lund University, SE-22100 Lund, Sweden.
authors.forename: Peter
authors.initials: P
authors.lastname: Jönsson
authors.affiliation: Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge, UK.
authors.forename: David
authors.initials: D
authors.lastname: Wirthensohn
authors.affiliation: Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge, UK.
authors.forename: Erwin
authors.initials: E
authors.lastname: De Genst
authors.affiliation: Laboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Brussel, Belgium.
authors.forename: Cécile
authors.initials: C
authors.lastname: Vincke
authors.affiliation: Laboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Brussel, Belgium.
authors.forename: Serge
authors.initials: S
authors.lastname: Muyldermans
authors.affiliation: Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge, UK.
authors.forename: Chris
authors.initials: C
authors.lastname: Dobson
authors.affiliation: Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge, UK.
authors.forename: David
authors.initials: D
authors.lastname: Klenerman
mesh_terms: D016229:Amyloid beta-Peptides / Q000138:chemical synthesis / Q000276:immunology / Q000378:metabolism*; D000818:Animals; D001253:Astrocytes / Q000166:cytology / Q000378:metabolism; D002118:Calcium / Q000378:metabolism*; D002232:Carbocyanines / Q000737:chemistry; D002462:Cell Membrane / Q000378:metabolism*; D002478:Cells, Cultured; D051152:Clusterin / Q000378:metabolism; D008856:Microscopy, Fluorescence; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000276:immunology / Q000378:metabolism*; D038761:Photobleaching; D066329:Protein Aggregates / Q000502:physiology*; D051381:Rats; D061905:Single-Domain Antibodies / Q000276:immunology
chemical_list: C569686:Alexa Fluor 647; D016229:Amyloid beta-Peptides; D002232:Carbocyanines; D051152:Clusterin; D010446:Peptide Fragments; D066329:Protein Aggregates; D061905:Single-Domain Antibodies; C075222:amyloid beta-protein (1-42); D002118:Calcium