Novel

Strategies in
Alzheimer’s
Disease
Treatment
PRESENTER: SCOTT RICHARDSON
MENTOR: DR. KRUGER
What is Alzheimer’s Disease

[1]
 Etiology

Amyloid Precursor Protein

N
AβP
C

Lysosomes Secretase

N AβP C N C

Amyloid
Normal Cellular
Functioning
Neurofibrillary Tangles Cell Death
Plaque and Tangles

[2]
Long Term Potentiation Inhibition

[3]
Cellular Prion Proteins (PrP)

[4]
 N1 Fragment

[5]
 Objectives

▪ The goal of this study was to:

1. Assess the role of PrPC in the cytotoxicity and LTP-inhibition of
primary neurons
2. Assess the use of recombinant PrP (rPrP) and N1 in the
cytotoxicity and LTP-inhibition of primary neurons
 Preparation of Aβ Oligomers

▪ Aβ-rPrP mixtures prepared in two ways:

[6]
Addition of PrPR to non-aggregated Addition of rPrP to aggregated
Aβ (preparation A) Aβ (preparation B)
 Neuronal PrPC expression

▪ Two types of Neurons used:

1. Neurons expressing PrPC
2. Neurons not expressing PrPC
▪ Knocked out with Prnp-siRNA or PrPC null mice were used
 Measurements

1. LTP-inhibition
▪ Slope of field evoked post-synaptic potentials (fEPSPs) from
mouse hippocampal slices
2. Cytotoxicity
▪ Measured with a lactate dehydrogenase (LDH) assay kit
Experimental Methods

Preparation of Aβ-rPrP mixtures

Addition of rPrP to Addition of rPrP to non-
aggregated Aβ mixtures aggregated mixtures

Addition of mixtures to neurons

Neurons expressing PrPC Neurons not expressing PrPC

Measurements
Atomic Force
LTP Cytotoxicity
Microscopy (AFM)
 Results: AFM with Preparation A

[6]

Aβ only Aβ with PrPr (5:1) Aβ with N-1 (5:1)

 Results: AFM with Preparation B

[6]
Aβ only Aβ with PrP Aβ with N1
 Results: LTP with Preparation A

Addition of rPrP to non-aggregated Aβ-mixtures Addition of N1 to non-aggregated Aβ-mixtures

Results: LTP with Preparation B

Addition of N1 of rPrP to aggregated Aβ-mixtures

Results: PrPC and LTP inhibition

Addition of Aβ to mice expressing PrP

or not expressing PrPC
Results: Cytotoxicity of Aβ with rPrP or N1

Preparation A Preparation B
Results: PrPC and LTP inhibition

Cytotoxicity of wild-type and PrP-null neurons with

preparation A (non-aggregated) or B (aggregated)
 Conclusion

▪ LTP inhibition proceeds via a PrPC dependent mechanism

▪ Cytotoxicity proceed via an PrPC independent mechanism
▪ rPrP and N1 show improvement in Aβ LTP inhibition
▪ rPrP and N1 show improvement in Aβ cell cytotoxicity
 Future Studies

▪ Peptide construction
▪ Pharmacology
▪ Active species of Aβ
 Thank you!

▪ Dr. Kreugir
▪ Dr. Striplin
▪ Dr. Merkert
 Citations

1. Long-term Potentiation. (2012, April 2). Retrieved from

http://www.expertsmind.com/topic/neuroscience/long-term-pot
entiation-93811.aspx
2. Huang, Ziwei, et al. "Proposed three-dimensional structure for
the cellular prion protein." Proceedings of the National Academy
of Sciences 91.15 (1994): 7139-7143.