{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,20]],"date-time":"2026-01-20T05:12:59Z","timestamp":1768885979630,"version":"3.49.0"},"reference-count":55,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2022,8,20]],"date-time":"2022-08-20T00:00:00Z","timestamp":1660953600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100006312","name":"BrightFocus Foundation","doi-asserted-by":"publisher","award":["A2019056F"],"award-info":[{"award-number":["A2019056F"]}],"id":[{"id":"10.13039\/100006312","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100006312","name":"BrightFocus Foundation","doi-asserted-by":"publisher","award":["AG 44486"],"award-info":[{"award-number":["AG 44486"]}],"id":[{"id":"10.13039\/100006312","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100006312","name":"BrightFocus Foundation","doi-asserted-by":"publisher","award":["AG 15379"],"award-info":[{"award-number":["AG 15379"]}],"id":[{"id":"10.13039\/100006312","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000049","name":"National Institute of Health","doi-asserted-by":"publisher","award":["A2019056F"],"award-info":[{"award-number":["A2019056F"]}],"id":[{"id":"10.13039\/100000049","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000049","name":"National Institute of Health","doi-asserted-by":"publisher","award":["AG 44486"],"award-info":[{"award-number":["AG 44486"]}],"id":[{"id":"10.13039\/100000049","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000049","name":"National Institute of Health","doi-asserted-by":"publisher","award":["AG 15379"],"award-info":[{"award-number":["AG 15379"]}],"id":[{"id":"10.13039\/100000049","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biosensors"],"abstract":"<jats:p>Amyloid-beta (A\u03b2) peptides are produced within neurons. Some peptides are released into the brain parenchyma, while others are retained inside the neurons. However, the detection of intracellular A\u03b2 remains a challenge since antibodies against A\u03b2 capture A\u03b2 and its precursor proteins (i.e., APP and C99). To overcome this drawback, we recently developed 1) the C99 720-670 biosensor for recording \u03b3-secretase activity and 2) a unique multiplexed immunostaining platform that enables the selective detection of intracellular A\u03b2 with subcellular resolution. Using these new assays, we showed that C99 is predominantly processed by \u03b3-secretase in late endosomes and lysosomes, and intracellular A\u03b2 is enriched in the same subcellular loci in intact neurons. However, the detailed properties of A\u03b2 in the acidic compartments remain unclear. Here, we report using fluorescent lifetime imaging microscopy (FLIM) that intracellular A\u03b2 includes both long A\u03b2 intermediates bound to \u03b3-secretase and short peptides dissociated from the protease complex. Surprisingly, our results also suggest that the dissociated A\u03b2 is bound to the glycoproteins on the inner membrane of lysosomes. Furthermore, we show striking cell-to-cell heterogeneity in intracellular A\u03b2 levels in primary neurons and APP transgenic mouse brains. These findings provide a basis for the further investigation of the role(s) of intracellular A\u03b2 and its relevance to Alzheimer\u2019s disease (AD).<\/jats:p>","DOI":"10.3390\/bios12080663","type":"journal-article","created":{"date-parts":[[2022,8,21]],"date-time":"2022-08-21T21:05:51Z","timestamp":1661115951000},"page":"663","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":17,"title":["In-Depth Characterization of Endo-Lysosomal A\u03b2 in Intact Neurons"],"prefix":"10.3390","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4996-4113","authenticated-orcid":false,"given":"Alec K.","family":"McKendell","sequence":"first","affiliation":[{"name":"Alzheimer Research Unit, MassGeneral Institute for Neurodegenerative Disease, Massachusetts General Hospital, Harvard Medical School, 114, 16th street, Charlestown, MA 02129, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mei C. Q.","family":"Houser","sequence":"additional","affiliation":[{"name":"Alzheimer Research Unit, MassGeneral Institute for Neurodegenerative Disease, Massachusetts General Hospital, Harvard Medical School, 114, 16th street, Charlestown, MA 02129, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shane P. C.","family":"Mitchell","sequence":"additional","affiliation":[{"name":"Alzheimer Research Unit, MassGeneral Institute for Neurodegenerative Disease, Massachusetts General Hospital, Harvard Medical School, 114, 16th street, Charlestown, MA 02129, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5721-9092","authenticated-orcid":false,"given":"Michael S.","family":"Wolfe","sequence":"additional","affiliation":[{"name":"Department of Medicinal Chemistry, University of Kansas, 1567 Irving Hill Rd, Lawrence, Kansas City, KS 66045, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Oksana","family":"Berezovska","sequence":"additional","affiliation":[{"name":"Alzheimer Research Unit, MassGeneral Institute for Neurodegenerative Disease, Massachusetts General Hospital, Harvard Medical School, 114, 16th street, Charlestown, MA 02129, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1970-2462","authenticated-orcid":false,"given":"Masato","family":"Maesako","sequence":"additional","affiliation":[{"name":"Alzheimer Research Unit, MassGeneral Institute for Neurodegenerative Disease, Massachusetts General Hospital, Harvard Medical School, 114, 16th street, Charlestown, MA 02129, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,8,20]]},"reference":[{"key":"ref_1","first-page":"CD001190","article-title":"Donepezil for dementia due to Alzheimer\u2019s disease","volume":"6","author":"Birks","year":"2018","journal-title":"Cochrane Database Syst. 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