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Cellular prion protein mediates impairment of synaptic plasticity by amyloid-beta oligomers.

Laurén J, Gimbel DA, Nygaard HB, Gilbert JW, Strittmatter SM - Nature (2009)

Bottom Line: Here we identify the cellular prion protein (PrP(C)) as an amyloid-beta-oligomer receptor by expression cloning.Amyloid-beta oligomers bind with nanomolar affinity to PrP(C), but the interaction does not require the infectious PrP(Sc) conformation.Anti-PrP antibodies prevent amyloid-beta-oligomer binding to PrP(C) and rescue synaptic plasticity in hippocampal slices from oligomeric amyloid-beta.

View Article: PubMed Central - PubMed

Affiliation: Cellular Neuroscience, Neurodegeneration and Repair Program, Yale University School of Medicine, New Haven, Connecticut 06536, USA.

ABSTRACT
A pathological hallmark of Alzheimer's disease is an accumulation of insoluble plaque containing the amyloid-beta peptide of 40-42 amino acid residues. Prefibrillar, soluble oligomers of amyloid-beta have been recognized to be early and key intermediates in Alzheimer's-disease-related synaptic dysfunction. At nanomolar concentrations, soluble amyloid-beta oligomers block hippocampal long-term potentiation, cause dendritic spine retraction from pyramidal cells and impair rodent spatial memory. Soluble amyloid-beta oligomers have been prepared from chemical syntheses, transfected cell culture supernatants, transgenic mouse brain and human Alzheimer's disease brain. Together, these data imply a high-affinity cell-surface receptor for soluble amyloid-beta oligomers on neurons-one that is central to the pathophysiological process in Alzheimer's disease. Here we identify the cellular prion protein (PrP(C)) as an amyloid-beta-oligomer receptor by expression cloning. Amyloid-beta oligomers bind with nanomolar affinity to PrP(C), but the interaction does not require the infectious PrP(Sc) conformation. Synaptic responsiveness in hippocampal slices from young adult PrP mice is normal, but the amyloid-beta oligomer blockade of long-term potentiation is absent. Anti-PrP antibodies prevent amyloid-beta-oligomer binding to PrP(C) and rescue synaptic plasticity in hippocampal slices from oligomeric amyloid-beta. Thus, PrP(C) is a mediator of amyloid-beta-oligomer-induced synaptic dysfunction, and PrP(C)-specific pharmaceuticals may have therapeutic potential for Alzheimer's disease.

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Oligomeric Aβ42 binds to neurons and to cells expressing PrPCa, Freshly prepared, oligomeric, or fibrillary preparations of Aβ42 were examined by transmission electron microscopy with negative staining. The arrows indicate globular oligomers in the middle segment and a fibril in the lower segment. Scale bar, 25 nm. b, Oligomeric Aβ42 peptide was analyzed by size exclusion chromatography, monitoring absorbance at 220 nm (black) and light scattering (red). The void volume (Vo) and elution of bovine serum albumin (BSA) from a separate run are shown. c, Oligomeric Aβ42 peptide (200 nM total peptide) binds to 21 DIV hippocampal neurons, whereas fresh Aβ42 (200 nM) does not. Bound biotin-Aβ42 was visualized by alkaline phosphatase conjugated streptavidin. d, Dose dependence of oligomeric Aβ42 binding to hippocampal neurons.e, The binding of 40 nM oligomeric or freshly prepared Aβ42 to COS-7 expressing PrPC. f, g, Fresh or oligomeric Aβ42 binding to PrPC-expressing COS-7 cells as a function of Aβ42 total concentration (monomer equivalent for oligomer preparations). Data are mean ± sem, and the Scatchard analysis is presented in g. Scale bars, 100 µm for c and e.
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Figure 1: Oligomeric Aβ42 binds to neurons and to cells expressing PrPCa, Freshly prepared, oligomeric, or fibrillary preparations of Aβ42 were examined by transmission electron microscopy with negative staining. The arrows indicate globular oligomers in the middle segment and a fibril in the lower segment. Scale bar, 25 nm. b, Oligomeric Aβ42 peptide was analyzed by size exclusion chromatography, monitoring absorbance at 220 nm (black) and light scattering (red). The void volume (Vo) and elution of bovine serum albumin (BSA) from a separate run are shown. c, Oligomeric Aβ42 peptide (200 nM total peptide) binds to 21 DIV hippocampal neurons, whereas fresh Aβ42 (200 nM) does not. Bound biotin-Aβ42 was visualized by alkaline phosphatase conjugated streptavidin. d, Dose dependence of oligomeric Aβ42 binding to hippocampal neurons.e, The binding of 40 nM oligomeric or freshly prepared Aβ42 to COS-7 expressing PrPC. f, g, Fresh or oligomeric Aβ42 binding to PrPC-expressing COS-7 cells as a function of Aβ42 total concentration (monomer equivalent for oligomer preparations). Data are mean ± sem, and the Scatchard analysis is presented in g. Scale bars, 100 µm for c and e.

Mentions: To characterize Aβ-oligomer binding sites, we synthesized biotin-Aβ42 peptide, denatured the peptide and allowed oligomers to form as described for ADDLs4. Consistent with findings for untagged Aβ42-oligomers5, biotin-Aβ42-oligomer preparations contain spherical particles of 5–6 nm diameter visible by negative staining in transmission electron microscopy, with rare protofibrils and no larger fibrils (Fig. 1a). Approximately 50% of peptide migrates by size exclusion chromatography (SEC) as a distinct assembly with a size of approximately 500 kDa corresponding to 50–100 Aβ monomers (Fig. 1b). Low molecular weight forms of Aβ42 in either oligomeric and fresh preparations migrate by SEC as monomers (Fig. 1b), demonstrating that the trimers or tetramers observed by SDS-PAGE (Suppl. Fig. 1) are not present under native conditions (and ref.10). Aβ42-oligomer binds to hippocampal neurons, whereas freshly prepared biotin-Aβ42 does not (Fig. 1c; Suppl. Fig. 2). Biotin-Aβ42-oligomer binding is enriched in MAP2-positive dendrites, with lower levels in βIII-tubulin positive axons, and very low levels in astroglial cells (Suppl. Fig. 3a, c, not shown and ref.6). The Aβ42-oligomer binding is most concentrated at post-synaptic densities marked by immunoreactive PSD-95 (Suppl. Fig. 3b). Binding to neurons is saturable, with an apparent KD of 50–100 nM monomer equivalent (Fig. 1d). The KD of the relevant Aβ42 assembly must be much less than 100 nM because minimal binding is detected with freshly prepared Aβ42. If the Aβ42 species responsible for binding contains 100 monomers and represents 50% of all biotin-Aβ42 in the preparation, the corrected affinity would be ∼0.4 nM. While this formulation of Aβ42-oligomer is not chromatographically identical to Aβ42-oligomer from brain2,3,9, it affords detection of high affinity binding sites likely to share pathological actions with sites for other Aβ42-oligomer preparations5,6,11.


Cellular prion protein mediates impairment of synaptic plasticity by amyloid-beta oligomers.

Laurén J, Gimbel DA, Nygaard HB, Gilbert JW, Strittmatter SM - Nature (2009)

Oligomeric Aβ42 binds to neurons and to cells expressing PrPCa, Freshly prepared, oligomeric, or fibrillary preparations of Aβ42 were examined by transmission electron microscopy with negative staining. The arrows indicate globular oligomers in the middle segment and a fibril in the lower segment. Scale bar, 25 nm. b, Oligomeric Aβ42 peptide was analyzed by size exclusion chromatography, monitoring absorbance at 220 nm (black) and light scattering (red). The void volume (Vo) and elution of bovine serum albumin (BSA) from a separate run are shown. c, Oligomeric Aβ42 peptide (200 nM total peptide) binds to 21 DIV hippocampal neurons, whereas fresh Aβ42 (200 nM) does not. Bound biotin-Aβ42 was visualized by alkaline phosphatase conjugated streptavidin. d, Dose dependence of oligomeric Aβ42 binding to hippocampal neurons.e, The binding of 40 nM oligomeric or freshly prepared Aβ42 to COS-7 expressing PrPC. f, g, Fresh or oligomeric Aβ42 binding to PrPC-expressing COS-7 cells as a function of Aβ42 total concentration (monomer equivalent for oligomer preparations). Data are mean ± sem, and the Scatchard analysis is presented in g. Scale bars, 100 µm for c and e.
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Figure 1: Oligomeric Aβ42 binds to neurons and to cells expressing PrPCa, Freshly prepared, oligomeric, or fibrillary preparations of Aβ42 were examined by transmission electron microscopy with negative staining. The arrows indicate globular oligomers in the middle segment and a fibril in the lower segment. Scale bar, 25 nm. b, Oligomeric Aβ42 peptide was analyzed by size exclusion chromatography, monitoring absorbance at 220 nm (black) and light scattering (red). The void volume (Vo) and elution of bovine serum albumin (BSA) from a separate run are shown. c, Oligomeric Aβ42 peptide (200 nM total peptide) binds to 21 DIV hippocampal neurons, whereas fresh Aβ42 (200 nM) does not. Bound biotin-Aβ42 was visualized by alkaline phosphatase conjugated streptavidin. d, Dose dependence of oligomeric Aβ42 binding to hippocampal neurons.e, The binding of 40 nM oligomeric or freshly prepared Aβ42 to COS-7 expressing PrPC. f, g, Fresh or oligomeric Aβ42 binding to PrPC-expressing COS-7 cells as a function of Aβ42 total concentration (monomer equivalent for oligomer preparations). Data are mean ± sem, and the Scatchard analysis is presented in g. Scale bars, 100 µm for c and e.
Mentions: To characterize Aβ-oligomer binding sites, we synthesized biotin-Aβ42 peptide, denatured the peptide and allowed oligomers to form as described for ADDLs4. Consistent with findings for untagged Aβ42-oligomers5, biotin-Aβ42-oligomer preparations contain spherical particles of 5–6 nm diameter visible by negative staining in transmission electron microscopy, with rare protofibrils and no larger fibrils (Fig. 1a). Approximately 50% of peptide migrates by size exclusion chromatography (SEC) as a distinct assembly with a size of approximately 500 kDa corresponding to 50–100 Aβ monomers (Fig. 1b). Low molecular weight forms of Aβ42 in either oligomeric and fresh preparations migrate by SEC as monomers (Fig. 1b), demonstrating that the trimers or tetramers observed by SDS-PAGE (Suppl. Fig. 1) are not present under native conditions (and ref.10). Aβ42-oligomer binds to hippocampal neurons, whereas freshly prepared biotin-Aβ42 does not (Fig. 1c; Suppl. Fig. 2). Biotin-Aβ42-oligomer binding is enriched in MAP2-positive dendrites, with lower levels in βIII-tubulin positive axons, and very low levels in astroglial cells (Suppl. Fig. 3a, c, not shown and ref.6). The Aβ42-oligomer binding is most concentrated at post-synaptic densities marked by immunoreactive PSD-95 (Suppl. Fig. 3b). Binding to neurons is saturable, with an apparent KD of 50–100 nM monomer equivalent (Fig. 1d). The KD of the relevant Aβ42 assembly must be much less than 100 nM because minimal binding is detected with freshly prepared Aβ42. If the Aβ42 species responsible for binding contains 100 monomers and represents 50% of all biotin-Aβ42 in the preparation, the corrected affinity would be ∼0.4 nM. While this formulation of Aβ42-oligomer is not chromatographically identical to Aβ42-oligomer from brain2,3,9, it affords detection of high affinity binding sites likely to share pathological actions with sites for other Aβ42-oligomer preparations5,6,11.

Bottom Line: Here we identify the cellular prion protein (PrP(C)) as an amyloid-beta-oligomer receptor by expression cloning.Amyloid-beta oligomers bind with nanomolar affinity to PrP(C), but the interaction does not require the infectious PrP(Sc) conformation.Anti-PrP antibodies prevent amyloid-beta-oligomer binding to PrP(C) and rescue synaptic plasticity in hippocampal slices from oligomeric amyloid-beta.

View Article: PubMed Central - PubMed

Affiliation: Cellular Neuroscience, Neurodegeneration and Repair Program, Yale University School of Medicine, New Haven, Connecticut 06536, USA.

ABSTRACT
A pathological hallmark of Alzheimer's disease is an accumulation of insoluble plaque containing the amyloid-beta peptide of 40-42 amino acid residues. Prefibrillar, soluble oligomers of amyloid-beta have been recognized to be early and key intermediates in Alzheimer's-disease-related synaptic dysfunction. At nanomolar concentrations, soluble amyloid-beta oligomers block hippocampal long-term potentiation, cause dendritic spine retraction from pyramidal cells and impair rodent spatial memory. Soluble amyloid-beta oligomers have been prepared from chemical syntheses, transfected cell culture supernatants, transgenic mouse brain and human Alzheimer's disease brain. Together, these data imply a high-affinity cell-surface receptor for soluble amyloid-beta oligomers on neurons-one that is central to the pathophysiological process in Alzheimer's disease. Here we identify the cellular prion protein (PrP(C)) as an amyloid-beta-oligomer receptor by expression cloning. Amyloid-beta oligomers bind with nanomolar affinity to PrP(C), but the interaction does not require the infectious PrP(Sc) conformation. Synaptic responsiveness in hippocampal slices from young adult PrP mice is normal, but the amyloid-beta oligomer blockade of long-term potentiation is absent. Anti-PrP antibodies prevent amyloid-beta-oligomer binding to PrP(C) and rescue synaptic plasticity in hippocampal slices from oligomeric amyloid-beta. Thus, PrP(C) is a mediator of amyloid-beta-oligomer-induced synaptic dysfunction, and PrP(C)-specific pharmaceuticals may have therapeutic potential for Alzheimer's disease.

Show MeSH
Related in: MedlinePlus