{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,18]],"date-time":"2026-07-18T23:05:46Z","timestamp":1784415946447,"version":"3.55.0"},"reference-count":48,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2024,1,21]],"date-time":"2024-01-21T00:00:00Z","timestamp":1705795200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Neuropathology Appl Neurobio"],"published-print":{"date-parts":[[2024,2]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Objective<\/jats:title>\n                    <jats:p>Filipin complex is an autooxidation\u2010prone fluorescent histochemical stain used in the diagnosis of Niemann\u2010Pick Disease Type C (NP\u2010C), a neurodegenerative lysosomal storage disorder. It is also widely used by researchers examining the distribution and accumulation of unesterified cholesterol in cell and animal models of neurodegenerative diseases including NP\u2010C and Sanfilippo syndrome (mucopolysaccharidosis IIIA; MPS IIIA). Recently, it has been suggested to be useful in studying Alzheimer's and Huntington's disease. Given filipin's susceptibility to photobleaching, we sought to establish a quantitative biochemical method for free cholesterol measurement.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>Brain tissue from mice with MPS IIIA was stained with filipin. Total and free cholesterol in brain homogenates was measured using a commercially available kit and a quantitative LC\u2013MS\/MS assay was developed. Gangliosides GM1, GM2 and GM3 were also quantified using LC\u2013MS\/MS.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>As anticipated, the MPS IIIA mouse brain displayed large numbers of filipin\u2010positive intra\u2010cytoplasmic inclusions, presumptively endo\u2010lysosomes. Challenging the prevailing dogma, however, we found no difference in the amount of free cholesterol in MPS IIIA mouse brain homogenates cf. control tissue, using either the fluorometric kit or LC\u2013MS\/MS assay. Filipin has previously been reported to bind to GM1 ganglioside, however, this lipid does not accumulate in MPS IIIA cells\/tissues. Using a fluorometric assay, we demonstrate for the first time that filipin cross\u2010reacts with both GM2 and GM3 gangliosides, explaining the filipin\u2010reactive inclusions observed in MPS IIIA brain cells.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>Filipin is not specific for free cholesterol, and positive staining in any setting should be interpreted with caution.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1111\/nan.12950","type":"journal-article","created":{"date-parts":[[2023,12,19]],"date-time":"2023-12-19T06:07:07Z","timestamp":1702966027000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Filipin complex\u2010reactive brain lesions: A cautionary tale"],"prefix":"10.1111","volume":"50","author":[{"given":"Adeline A.","family":"Lau","sequence":"first","affiliation":[{"name":"Childhood Dementia Research Group, Flinders Health and Medical Research Institute Flinders University  Bedford Park South Australia Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paul J.","family":"Trim","sequence":"additional","affiliation":[{"name":"Proteomics, Metabolomics and MS\u2010Imaging Core Facility South Australian Health and Medical Research Institute  Adelaide South Australia Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Barbara M.","family":"King","sequence":"additional","affiliation":[{"name":"Childhood Dementia Research Group, Flinders Health and Medical Research Institute Flinders University  Bedford Park South Australia Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sofia","family":"Hassiotis","sequence":"additional","affiliation":[{"name":"Childhood Dementia Research Group, Flinders Health and Medical Research Institute Flinders University  Bedford Park South Australia Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ya Hui","family":"Hung","sequence":"additional","affiliation":[{"name":"Oxidation Biology Unit Florey Institute of Neuroscience and Mental Health  Parkville Victoria Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ashley I.","family":"Bush","sequence":"additional","affiliation":[{"name":"Oxidation Biology Unit Florey Institute of Neuroscience and Mental Health  Parkville Victoria Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marten F.","family":"Snel","sequence":"additional","affiliation":[{"name":"Proteomics, Metabolomics and MS\u2010Imaging Core Facility South Australian Health and Medical Research Institute  Adelaide South Australia Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1038-9884","authenticated-orcid":false,"given":"Kim 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