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Purification and characterization of astrocyte-secreted apolipoprotein E and J-containing lipoproteins from wild-type and human apoE transgenic mice
Abstract
The varepsilon4 allele of apolipoprotein E (apoE) is a genetic risk factor for Alzheimer's disease (AD). In order to gain a better understanding of the molecular mechanisms by which apoE and possibly other apolipoproteins produced in the central nervous system (CNS) influence AD pathogenesis, we have purified and characterized the two most abundant apolipoproteins produced in the CNS, apoE and apoJ. We purified apoE and apoJ from primary cultures of mouse astrocytes, which were derived from transgenic mice expressing human apoE isoforms in the absence of mouse apoE. Utilizing antibody affinity columns, we were able to purify both human apoE3 and apoE4, as well as mouse apoJ-containing lipoproteins. Astrocyte-secreted human apoE was present in high density-like lipoproteins of three predominant sizes ranging from 8 to 15 nm in diameter. Mouse apoJ was in particles between 10 and 17 nm in diameter with a peak size range of approximately 11 nm. ApoE and apoJ were in distinct lipoproteins. Utilization of quick-freeze, deep-etch electron microscopy revealed the apoE particles were discs while the apoJ particles were smaller and more irregular in appearance. The lipid composition of apoE particles was very different from those containing apoJ. ApoE-particles contained a similar mass of apoE and lipid, with cholesterol and phospholipid being about equal in mass per particle. ApoJ-particles were relatively lipid poor (three parts protein, one part lipid), with phospholipids being much more abundant than cholesterol. Detailed characterization of phospholipid composition by electrospray ionization mass spectrometry analysis revealed ethanolamine glycerophospholipids to be the most abundant phospholipid present in both apoE and apoJ particles. Analysis of cerebrospinal fluid from apoE3 and apoE4 transgenic mice revealed that human and mouse apoE were in particles the same size as those secreted by astrocytes. Further use of physiological preparations of CNS-derived lipoproteins may allow for a detailed understanding of the role of these molecules in the normal brain and in diseases such as AD.
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In the citing paper
“Astrocytes secrete ApoE in a similar size ranging from 8 to 20 nm [17, 19, 20].”
From ApoE lipoproteins in the central nervous system under homeostasis and role in Alzheimer’s disease and related disorders (2026) · cites this paper as [19]
In the citing paper
“Under homeostatic conditions, ApoE is primarily produced by astrocytes which secrete lipidated ApoE [17–20].”
From ApoE lipoproteins in the central nervous system under homeostasis and role in Alzheimer’s disease and related disorders (2026) · cites this paper as [19]
In the citing paper
“Astrocytic LpE was found to be enriched in proteins such as phospholipid transfer protein (PLTP) and clusterin (Clu, also known as ApoJ), both of which play a role in AD disease pathogenesis (Fig. 4A) [11, 19, 101].”
From ApoE lipoproteins in the central nervous system under homeostasis and role in Alzheimer’s disease and related disorders (2026) · cites this paper as [19]
In the citing paper
“In the central nervous system (CNS), ApoE is primarily secreted by astrocytes under homeostatic conditions with microglia upregulating ApoE secretion under multiple disease conditions [18–20, 93–95].”
From ApoE lipoproteins in the central nervous system under homeostasis and role in Alzheimer’s disease and related disorders (2026) · cites this paper as [19]
In the citing paper
“Clu, which is secreted separately from ApoE-containing lipoproteins but can be found together with ApoE in CSF lipoproteins, was also identified as a risk factor for AD in a genome wide association study [19, 103, 104].”
From ApoE lipoproteins in the central nervous system under homeostasis and role in Alzheimer’s disease and related disorders (2026) · cites this paper as [19]
In the citing paper
“Astrocytic LpE was found to be enriched in proteins such as phospholipid transfer protein (PLTP) and clusterin (Clu, also known as ApoJ), both of which play a role in AD disease pathogenesis (Fig. 4A) [11, 19, 101].”
From ApoE lipoproteins in the central nervous system under homeostasis and role in Alzheimer’s disease and related disorders (2026) · cites this paper as [19]
In the citing paper
“Clu, which is secreted separately from ApoE-containing lipoproteins but can be found together with ApoE in CSF lipoproteins, was also identified as a risk factor for AD in a genome wide association study [19, 103, 104].”
From ApoE lipoproteins in the central nervous system under homeostasis and role in Alzheimer’s disease and related disorders (2026) · cites this paper as [19]
In the citing paper
“Astrocytes with human APOE isoforms knocked into the endogenous mouse Apoe allele secrete discoidal particles of overlapping size.”
From ApoE lipoproteins in the central nervous system under homeostasis and role in Alzheimer’s disease and related disorders (2026) · cites this paper as [19]
In the citing paper
“However, ApoE2 lipoproteins secreted from astrocytes form slightly larger particles than ApoE3 and ApoE4 lipoproteins [17–20].”
From ApoE lipoproteins in the central nervous system under homeostasis and role in Alzheimer’s disease and related disorders (2026) · cites this paper as [19]
In the citing paper
“lipids that are normally of relatively low abundance in membranes and ApoE- and ApoJ-containing lipoparticles14”
From Neurotoxic reactive astrocytes induce cell death via saturated lipids (2021) · cites this paper as [14]
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Metadata
- Authors
- DeMattos, R. B., Brendza, R. P., Heuser, J. E., Kierson, M., Cirrito, J. R., Fryer, J., Sullivan, P. M., Fagan, A. M., Han, X., & Holtzman, D. M.
- Affiliations
- Washington University in St. Louis (US)
- Duke University (US)
- Corresponding author
- Holtzman, David M
- ORCID
- Venue
- Neurochemistry International
- Publisher
- Elsevier BV
- Published
- 2001-11
- Volume
- 39
- Issue
- 5-6
- Pages
- 415-425
- ISSN
- 0197-0186 (ISSN-L)
- OpenAlex
- W2017404389
- Cited by
- 180
- References
- 74
- Citations by year
- 2026: 4 · 2025: 8 · 2024: 6 · 2023: 6 · 2022: 5 · 2021: 6 · 2020: 7 · 2019: 10 · 2018: 7 · 2017: 6 · 2016: 13 · 2015: 6 · 2014: 10 · 2013: 6 · 2012: 8
- FWCI
- 5.19
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Neurochemistry International (publishedVersion)
- PubMed (publishedVersion)
- Funders
- James S. McDonnell Foundation
- National Institutes of Health
- National Institute on Aging (AG00861, AG 05681, P50AG005681, AG13956)
- National Institute of General Medical Sciences (GM29647, R01GM029647)
- Topics
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Alzheimer's disease research and treatments
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biochemistry › Cholesterol and Lipid Metabolism
- Health Sciences › Medicine › Cardiology and Cardiovascular Medicine › Lipoproteins and Cardiovascular Health
- Keywords
- apolipoprotein E, clusterin, astrocytes, lipoprotein secretion, APOE ε4, APOE3, Alzheimer's disease, high-density lipoprotein, lipoprotein particle size, phospholipid composition, ethanolamine glycerophospholipids, cerebrospinal fluid
- MeSH terms
- Animals, Apolipoproteins E, Astrocytes, Cells, Cultured, Glycoproteins, Humans, Lipids, Lipoproteins, Lipoproteins, HDL, Mice, Transgenic, Particle Size, Phospholipids, Reference Values, Molecular Chaperones, Clusterin, Mice
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