Expression of the Adrenoleukodystrophy Protein in the Human and Mouse Central Nervous System

Fouquet, Françoise; Zhou, Jia Min; Ralston, Evelyn; Murray, Kerren; Troalen, Frédéric; Magal, Ella; Robain, O.; Dubois‐Dalcq, Monique; Aubourg, Patrick

doi:10.1006/nbdi.1997.0127

Cited by 100 publications

(65 citation statements)

References 54 publications

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“…Nuclei were removed by centrifugation for 10 min at 4°C and 1,000 ϫ g. The cell lysate was incubated twice with agarose beads for 1 h at 4°C with gentle rotation. After this pre-clearing step, lysate was incubated with rabbit polyclonal anti-ALDP antibody (22) cross-linked to agarose beads via dimethyl pimelimidate (protocol adapted from Ref. 23) or mouse monoclonal anti-GFP antibody covalently immobilized on agarose beads (MBL) for 1 h at 4°C with gentle rotation.…”

Section: Methodsmentioning

confidence: 99%

“…The membrane was first incubated in 5% skimmed milk dissolved in PBS, 0.1% Tween 20 (PBS/T) for 1 h at room temperature. The membrane was probed with 1/500 diluted mouse anti-GFP antibody (Roche Applied Science) or with rabbit polyclonal anti-ALDP (1/5,000) antibody (22) and then with the appropriate secondary antibody coupled to horseradish peroxidase (1/5,000 Santa Cruz Biotechnology), before being revealed by ECL (Santa Cruz Biotechnology).…”

Section: Methodsmentioning

confidence: 99%

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Substrate Specificity Overlap and Interaction between Adrenoleukodystrophy Protein (ALDP/ABCD1) and Adrenoleukodystrophy-related Protein (ALDRP/ABCD2)

Génin

Geillon

Gondcaille

et al. 2011

Journal of Biological Chemistry

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X-linked adrenoleukodystrophy (X-ALD) is a neurodegenerative disorder caused by mutations in the ABCD1 gene, which encodes a peroxisomal member of the ATP-binding cassette (ABC) transporter subfamily D called ALDP. ALDP is supposed to function as a homodimer allowing the entry of CoA-esters of very-long chain fatty acids (VLCFA) into the peroxisome, the unique site of their ␤-oxidation. ALDP deficiency can be corrected by overexpression of ALDRP, its closest homolog. However, the exact nature of the substrates transported by ALDRP and its relationships with ALDP still remain unclear. To gain insight into the function of ALDRP, we used cell models allowing the induction in a dose-dependent manner of a wild type or a mutated non-functional ALDRP-EGFP fusion protein. We explored the consequences of the changes of ALDRP expression levels on the fatty acid content (saturated, monounsaturated, and polyunsaturated fatty acids) in phospholipids as well as on the levels of ␤-oxidation of 3 suspected substrates: C26:0, C24:0, and C22:6n-3 (DHA). We found an inverse correlation between the fatty acid content of saturated (C26:0, C24:0) and monounsaturated (C26:1, C24:1) VLCFA and the expression level of ALDRP. Interestingly, we obtained a transdominant-negative effect of the inactive ALDRP-EGFP on ALDP function. This effect is due to a physical interaction between ALDRP and ALDP that we evidenced by proximity ligation assays and coimmunoprecipitation. Finally, the ␤-oxidation assays demonstrate a role of ALDRP in the metabolism of saturated VLCFA (redundant with that of ALDP) but also a specific involvement of ALDRP in the metabolism of DHA. X-linked adrenoleukodystrophy (X-ALD)2 (OMIM 300100) is a very complex neurodegenerative disorder whose physiopathogenesis remains to be clarified (1). X-ALD is caused by mutations in the ABCD1 gene located in Xq28, which encodes a peroxisomal member of the ATP-binding cassette (ABC) transporter subfamily D called ALDP (adrenoleukodystrophy-protein) (2). This protein has the structure of a half ABC transporter, which is supposed to function as a homodimer (3, 4). However, heterodimerization with one of the other members of the ABCD subfamily (i.e. ALDRP (encoded by the ABCD2 gene (5)), PMP70 (ABCD3 (6)), and PMP69 (ABCD4 (7, 8)) cannot be excluded especially in the situation where these proteins are overexpressed. Although a mirror expression pattern is often observed between ALDP and ALDRP when specific cell types are analyzed (9), peroxisomal ABC transporters have overlapping expression patterns rendering possible such interactions (5, 10). Coimmunoprecipitation experiments or FRET analysis have demonstrated heterodimerization in cells overexpressing the peroxisomal ABC transporters (11,12). Although the peroxisomal localization of ALDP, ALDRP, and PMP70 is clearly demonstrated, PMP69 has recently been described to be localized in the endoplasmic reticulum and was found to be absent in the peroxisome. This excludes a possible interaction at the peroxisomal membrane with the other ABC...

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Substrate Specificity Overlap and Interaction between Adrenoleukodystrophy Protein (ALDP/ABCD1) and Adrenoleukodystrophy-related Protein (ALDRP/ABCD2)

Génin

Geillon

Gondcaille

et al. 2011

Journal of Biological Chemistry

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“…ABCD1 is not uniformly expressed across different cell types 9. How can axons be affected when ABCD1 is only expressed in neurons at very low levels?9 Microglia have recently been implicated as potential cellular mediators of synapse loss 10.…”

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confidence: 99%

Microglial dysfunction as a key pathological change in adrenomyeloneuropathy

Gong

Sasidharan

Laheji

et al. 2017

Annals of Neurology

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ObjectiveMutations in ABCD1 cause the neurodegenerative disease, adrenoleukodystrophy, which manifests as the spinal cord axonopathy adrenomyeloneuropathy (AMN) in nearly all males surviving into adulthood. Microglial dysfunction has long been implicated in pathogenesis of brain disease, but its role in the spinal cord is unclear.MethodsWe assessed spinal cord microglia in humans and mice with AMN and investigated the role of ABCD1 in microglial activity toward neuronal phagocytosis in cell culture. Because mutations in ABCD1 lead to incorporation of very‐long‐chain fatty acids into phospholipids, we separately examined the effects of lysophosphatidylcholine (LPC) upon microglia.ResultsWithin the spinal cord of humans and mice with AMN, upregulation of several phagocytosis‐related markers, such as MFGE8 and TREM2, precedes complement activation and synapse loss. Unexpectedly, this occurs in the absence of overt inflammation. LPC C26:0 added to ABCD1‐deficient microglia in culture further enhances MFGE8 expression, aggravates phagocytosis, and leads to neuronal injury. Furthermore, exposure to a MFGE8‐blocking antibody reduces phagocytic activity.InterpretationSpinal cord microglia lacking ABCD1 are primed for phagocytosis, affecting neurons within an altered metabolic milieu. Blocking phagocytosis or specific phagocytic receptors may alleviate synapse loss and axonal degeneration. Ann Neurol 2017;82:813–827

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“…As a result, VLCFAs accumulate in all tissues and plasma (8). In brain, ALDP is expressed in all glial cell types, including astrocytes, microglial cells, and oligodendrocytes (9). Myelinated tracts of oligodendrocytes have a high lipid turnover.…”

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confidence: 99%

Characterization of the human ω‐oxidation pathway for ω‐hydroxy‐very‐long‐chain fatty acids

Sanders

Ofman

Dacremont³

et al. 2008

FASEB j.

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Very-long-chain fatty acids (VLCFAs) have long been known to be degraded exclusively in peroxisomes via ␤-oxidation. A defect in peroxisomal ␤-oxidation results in elevated levels of VLCFAs and is associated with the most frequent inherited disorder of the central nervous system white matter, X-linked adrenoleukodystrophy. Recently, we demonstrated that VLCFAs can also undergo -oxidation, which may provide an alternative route for the breakdown of VLCFAs. The -oxidation of VLCFA is initiated by CYP4F2 and CYP4F3B, which produce -hydroxy-VLCFAs. In this article, we characterized the enzymes involved in the formation of very-long-chain dicarboxylic acids from -hydroxy-VLCFAs. We demonstrate that very-longchain dicarboxylic acids are produced via two independent pathways. The first is mediated by an as yet unidentified, microsomal NAD ؉ -dependent alcohol dehydrogenase and fatty aldehyde dehydrogenase, which is encoded by the ALDH3A2 gene and is deficient in patients with Sjögren-Larsson syndrome. The second pathway involves the NADPH-dependent hydroxylation of -hydroxy-VLCFAs by CYP4F2, CYP4F3B, or CYP4F3A. Enzyme kinetic studies show that oxidation of -hydroxy-VLCFAs occurs predominantly via the NAD ؉ -dependent route. Overall, our data demonstrate that in humans all enzymes are present for the complete conversion of VLCFAs to their corresponding very-long-chain dicarboxylic acids.-Sanders, R.-J., Ofman, R., Dacremont, G., Wanders, R. J. A., Kemp, S. Characterization of the human -oxidation pathway for -hydroxy-very-long-chain fatty acids. FASEB J. 22,

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Expression of the Adrenoleukodystrophy Protein in the Human and Mouse Central Nervous System

Cited by 100 publications

References 54 publications

Substrate Specificity Overlap and Interaction between Adrenoleukodystrophy Protein (ALDP/ABCD1) and Adrenoleukodystrophy-related Protein (ALDRP/ABCD2)

Substrate Specificity Overlap and Interaction between Adrenoleukodystrophy Protein (ALDP/ABCD1) and Adrenoleukodystrophy-related Protein (ALDRP/ABCD2)

Microglial dysfunction as a key pathological change in adrenomyeloneuropathy

Characterization of the human ω‐oxidation pathway for ω‐hydroxy‐very‐long‐chain fatty acids

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