In vivo measurement of apolipoprotein E from the brain interstitial fluid using microdialysis.

Jason D Ulrich,Gary E Landreth,Hong Jiang,Dorothy R Schuler,David M Holtzman,Thomas E Mahan,Philip B Verghese,Joseph M Castellano,Jessica L Restivo,Jack M Burchett,John R Cirrito

doi:10.1186/1750-1326-8-13

Abstract

BackgroundThe APOE4 allele variant is the strongest known genetic risk factor for developing late-onset Alzheimer’s disease. The link between apolipoprotein E (apoE) and Alzheimer’s disease is likely due in large part to the impact of apoE on the metabolism of amyloid β (Aβ) within the brain. Manipulation of apoE levels and lipidation within the brain has been proposed as a therapeutic target for the treatment of Alzheimer’s disease. However, we know little about the dynamic regulation of apoE levels and lipidation within the central nervous system. We have developed an assay to measure apoE levels in the brain interstitial fluid of awake and freely moving mice using large molecular weight cut-off microdialysis probes.ResultsWe were able to recover apoE using microdialysis from human cerebrospinal fluid (CSF) in vitro and mouse brain parenchyma in vivo. Microdialysis probes were inserted into the hippocampus of wild-type mice and interstitial fluid was collected for 36 hours. Levels of apoE within the microdialysis samples were determined by ELISA. The levels of apoE were found to be relatively stable over 36 hours. No apoE was detected in microdialysis samples from apoE KO mice. Administration of the RXR agonist bexarotene increased ISF apoE levels while ISF Aβ levels were decreased. Extrapolation to zero-flow analysis allowed us to determine the absolute recoverable concentration of apoE3 in the brain ISF of apoE3 KI mice. Furthermore, analysis of microdialysis samples by non-denaturing gel electrophoresis determined lipidated apoE particles in microdialysis samples were consistent in size with apoE particles from CSF. Finally, we found that the concentration of apoE in the brain ISF was dependent upon apoE isoform in human apoE KI mice, following the pattern apoE2>apoE3>apoE4.ConclusionsWe are able to collect lipidated apoE from the brain of awake and freely moving mice and monitor apoE levels over the course of several hours from a single mouse. Our technique enables assessment of brain apoE dynamics under physiological and pathophysiological conditions and in response to therapeutic interventions designed to affect apoE levels and lipidation within the brain.

Highlights

The APOE4 allele variant is the strongest known genetic risk factor for developing late-onset Alzheimer’s disease
We optimized the use of a 1,000 kDa molecular weight cut-off (MWCO) microdialysis probe for collection of apolipoprotein E (apoE)
As expected, decreasing the flow rate increased the amount of apoE detected in microdialysis samples (Figure 1A)

Summary

Introduction

The APOE4 allele variant is the strongest known genetic risk factor for developing late-onset Alzheimer’s disease. The link between apolipoprotein E (apoE) and Alzheimer’s disease is likely due in large part to the impact of apoE on the metabolism of amyloid β (Aβ) within the brain. Manipulation of apoE levels and lipidation within the brain has been proposed as a therapeutic target for the treatment of Alzheimer’s disease. Possession of the ε4 allele is the strongest known genetic risk factor for developing late-onset Alzheimer’s disease (LOAD), while apoE2 is protective[2]. ApoE likely influences AD in large part through the isoform-dependent effects of apoE on the metabolism of amyloid-β (Aβ) in the brain. ApoE likely plays additional roles in the brain in processes such as synaptic plasticity, regulation of extracellular lipid metabolism, and inflammation (for review, see [2])

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Results

Conclusion