{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T14:42:00Z","timestamp":1790779320531,"version":"4.1.0"},"reference-count":26,"publisher":"Oxford University Press (OUP)","issue":"2","license":[{"start":{"date-parts":[[2023,3,16]],"date-time":"2023-03-16T00:00:00Z","timestamp":1678924800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2023,4,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>Excessive cholesterol absorption from intestinal lumen contributes to the pathogenesis of hypercholesterolemia, which is an independent risk factor for atherosclerotic cardiovascular disease. Niemann-Pick C1-like 1 (NPC1L1) is a major membrane protein responsible for cholesterol absorption, in which the physiological role of vesicular endocytosis is still controversial, and it lacks a feasible tool to visualize and evaluate the endocytosis of NPC1L1 vesicles in vivo. Here, we genetically labeled endogenous NPC1L1 protein with EGFP in a knock-in mouse model, and demonstrated fluorescent visualization and evaluation of the endocytic vesicles of NPC1L1-cago during intestinal cholesterol absorption. The homozygous NPC1L1-EGFP mice have normal NPC1L1 expression pattern as well as cholesterol homeostasis on chow or high-cholesterol diets. The fluorescence of NPC1L1-EGFP fusion protein localizes at the brush border membrane of small intestine, and EGFP-positive vesicles is visualized beneath the membrane as early as 5 min post oral gavage of cholesterol. Of note, the vesicles colocalize with the early endosomal marker early endosome antigen 1 (EEA1) and the filipin-stained free cholesterol. Pretreatment with NPC1L1 inhibitor ezetimibe inhibits the formation of these cholesterol-induced endocytic vesicles. Our data support the notion that NPC1L1-mediated cholesterol absorption is a vesicular endocytic process. NPC1L1-EGFP mice are a useful model for visualizing cellular NPC1L1-cargo vesicle itineraries and for evaluating NPC1L1 activity in vivo in response to diverse pharmacological agents and nutrients.<\/jats:p>","DOI":"10.1093\/lifemeta\/load011","type":"journal-article","created":{"date-parts":[[2023,3,16]],"date-time":"2023-03-16T19:55:58Z","timestamp":1678996558000},"source":"Crossref","is-referenced-by-count":7,"title":["Fluorescent visualization and evaluation of NPC1L1-mediated vesicular endocytosis during intestinal cholesterol absorption in mice"],"prefix":"10.1093","volume":"2","author":[{"given":"Xiaojing","family":"Wu","sequence":"first","affiliation":[{"name":"Department of Pathophysiology, Naval Medical University , Shanghai 200433 , China"},{"name":"NHC Key Laboratory of Hormones and Development, Chu Hsien-I Memorial Hospital & Tianjin Institute of Endocrinology, Tianjin Medical University , Tianjin 300134 , China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xian-Hua","family":"Ma","sequence":"additional","affiliation":[{"name":"Department of Pathophysiology, Naval Medical University , Shanghai 200433 , China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jie","family":"Lin","sequence":"additional","affiliation":[{"name":"Department of Pathophysiology, Naval Medical University , Shanghai 200433 , China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaohang","family":"Yang","sequence":"additional","affiliation":[{"name":"NHC Key Laboratory of Hormones and Development, Chu Hsien-I Memorial Hospital & Tianjin Institute of Endocrinology, Tianjin Medical University , Tianjin 300134 , China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jian-Hui","family":"Shi","sequence":"additional","affiliation":[{"name":"Department of Pathophysiology, Naval Medical University , Shanghai 200433 , China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhifang","family":"Xie","sequence":"additional","affiliation":[{"name":"Ministry of Education Shanghai Key Laboratory of Children\u2019s Environmental Health, Xinhua Hospital, Shanghai Jiaotong University School of Medicine , Shanghai 200092 , China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yu-Xia","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Pathophysiology, Naval Medical University , Shanghai 200433 , China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Weiping J","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Pathophysiology, Naval Medical University , Shanghai 200433 , China"},{"name":"NHC Key Laboratory of Hormones and Development, Chu Hsien-I Memorial Hospital & Tianjin Institute of Endocrinology, Tianjin Medical University , Tianjin 300134 , China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2023,3,16]]},"reference":[{"key":"2023053006052186200_CIT0001","doi-asserted-by":"crossref","first-page":"225","DOI":"10.1038\/s41580-019-0190-7","article-title":"Mechanisms and regulation of cholesterol homeostasis","volume":"21","author":"Luo","year":"2020","journal-title":"Nat Rev Mol Cell Biol"},{"key":"2023053006052186200_CIT0002","doi-asserted-by":"crossref","first-page":"e56","DOI":"10.1161\/CIR.0000000000000659","article-title":"Heart disease and stroke statistics-2019 update: a report from the American Heart Association","volume":"139","author":"Benjamin","year":"2019","journal-title":"Circulation"},{"key":"2023053006052186200_CIT0003","doi-asserted-by":"crossref","first-page":"4","DOI":"10.1016\/j.addr.2020.07.019","article-title":"Lipoproteins and lipids in cardiovascular disease: from mechanistic insights to therapeutic targeting","volume":"159","author":"Soppert","year":"2020","journal-title":"Adv Drug Deliv Rev"},{"key":"2023053006052186200_CIT0004","doi-asserted-by":"crossref","first-page":"2459","DOI":"10.1093\/eurheartj\/ehx144","article-title":"Low-density lipoproteins cause atherosclerotic cardiovascular disease. 1. Evidence from genetic, epidemiologic, and clinical studies. A consensus statement from the European Atherosclerosis Society Consensus Panel","volume":"38","author":"Ference","year":"2017","journal-title":"Eur Heart J"},{"key":"2023053006052186200_CIT0005","doi-asserted-by":"crossref","first-page":"1201","DOI":"10.1126\/science.1093131","article-title":"Niemann-Pick C1 Like 1 protein is critical for intestinal cholesterol absorption","volume":"303","author":"Altmann","year":"2004","journal-title":"Science"},{"key":"2023053006052186200_CIT0006","doi-asserted-by":"crossref","first-page":"12710","DOI":"10.1074\/jbc.M409110200","article-title":"Inactivation of NPC1L1 causes multiple lipid transport defects and protects against diet-induced hypercholesterolemia","volume":"280","author":"Davies","year":"2005","journal-title":"J Biol Chem"},{"key":"2023053006052186200_CIT0007","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1006\/geno.2000.6151","article-title":"Evidence for a Niemann-pick C (NPC) gene family: identification and characterization of NPC1L1","volume":"65","author":"Davies","year":"2000","journal-title":"Genomics"},{"key":"2023053006052186200_CIT0008","doi-asserted-by":"crossref","first-page":"1653","DOI":"10.1194\/jlr.M800669-JLR200","article-title":"Membrane topology of human NPC1L1, a key protein in enterohepatic cholesterol absorption","volume":"50","author":"Wang","year":"2009","journal-title":"J Lipid Res"},{"key":"2023053006052186200_CIT0009","doi-asserted-by":"crossref","first-page":"25088","DOI":"10.1074\/jbc.M111.244475","article-title":"The N-terminal domain of NPC1L1 protein binds cholesterol and plays essential roles in cholesterol uptake","volume":"286","author":"Zhang","year":"2011","journal-title":"J Biol Chem"},{"key":"2023053006052186200_CIT0010","doi-asserted-by":"crossref","first-page":"80","DOI":"10.1038\/nm.3417","article-title":"The clathrin adaptor Numb regulates intestinal cholesterol absorption through dynamic interaction with NPC1L1","volume":"20","author":"Li","year":"2014","journal-title":"Nat Med"},{"key":"2023053006052186200_CIT0011","doi-asserted-by":"crossref","first-page":"2092","DOI":"10.1194\/jlr.M027359","article-title":"Ezetimibe blocks the internalization of NPC1L1 and cholesterol in mouse small intestine","volume":"53","author":"Xie \u8c22\u7545","year":"2012","journal-title":"J Lipid Res"},{"key":"2023053006052186200_CIT0012","doi-asserted-by":"crossref","first-page":"1968","DOI":"10.1172\/JCI30060","article-title":"Hepatic Niemann-Pick C1-like 1 regulates biliary cholesterol concentration and is a target of ezetimibe","volume":"117","author":"Temel","year":"2007","journal-title":"J Clin Invest"},{"key":"2023053006052186200_CIT0013","doi-asserted-by":"crossref","first-page":"G369","DOI":"10.1152\/ajpgi.00306.2006","article-title":"Modulation of human Niemann-Pick C1-like 1 gene expression by sterol: role of sterol regulatory element binding protein 2","volume":"292","author":"Alrefai","year":"2007","journal-title":"Am J Physiol Gastrointest Liver Physiol"},{"key":"2023053006052186200_CIT0014","doi-asserted-by":"crossref","first-page":"1354","DOI":"10.1194\/jlr.M900274-JLR200","article-title":"HNF1alpha and SREBP2 are important regulators of NPC1L1 in human liver","volume":"51","author":"Pramfalk","year":"2010","journal-title":"J Lipid Res"},{"key":"2023053006052186200_CIT0015","doi-asserted-by":"crossref","first-page":"6616","DOI":"10.1074\/jbc.M511123200","article-title":"Cholesterol-regulated translocation of NPC1L1 to the cell surface facilitates free cholesterol uptake","volume":"281","author":"Yu","year":"2006","journal-title":"J Biol Chem"},{"key":"2023053006052186200_CIT0016","doi-asserted-by":"crossref","first-page":"239","DOI":"10.1146\/annurev-physiol-012110-142233","article-title":"Niemann-pick C1-like 1 (NPC1L1) protein in intestinal and hepatic cholesterol transport","volume":"73","author":"Jia","year":"2011","journal-title":"Annu Rev Physiol"},{"key":"2023053006052186200_CIT0017","doi-asserted-by":"crossref","first-page":"508","DOI":"10.1016\/j.cmet.2008.04.001","article-title":"The cholesterol absorption inhibitor ezetimibe acts by blocking the sterol-induced internalization of NPC1L1","volume":"7","author":"Ge","year":"2008","journal-title":"Cell Metab"},{"key":"2023053006052186200_CIT0018","doi-asserted-by":"crossref","first-page":"1087","DOI":"10.1126\/science.aao6575","article-title":"A LIMA1 variant promotes low plasma LDL cholesterol and decreases intestinal cholesterol absorption","volume":"360","author":"Zhang","year":"2018","journal-title":"Science"},{"key":"2023053006052186200_CIT0019","doi-asserted-by":"crossref","first-page":"35933","DOI":"10.1074\/jbc.M111.270199","article-title":"The small GTPase Cdc42 interacts with Niemann-Pick C1-like 1 (NPC1L1) and controls its movement from endocytic recycling compartment to plasma membrane in a cholesterol-dependent manner","volume":"286","author":"Xie","year":"2011","journal-title":"J Biol Chem"},{"key":"2023053006052186200_CIT0020","doi-asserted-by":"crossref","first-page":"551","DOI":"10.1073\/pnas.1014434108","article-title":"Flotillins play an essential role in Niemann-Pick C1-like 1-mediated cholesterol uptake","volume":"108","author":"Ge","year":"2011","journal-title":"Proc Natl Acad Sci U S A"},{"key":"2023053006052186200_CIT0021","doi-asserted-by":"crossref","first-page":"8132","DOI":"10.1073\/pnas.0500269102","article-title":"The target of ezetimibe is Niemann-Pick C1-Like 1 (NPC1L1)","volume":"102","author":"Garcia-Calvo","year":"2005","journal-title":"Proc Natl Acad Sci U S A"},{"key":"2023053006052186200_CIT0022","doi-asserted-by":"crossref","first-page":"11140","DOI":"10.1073\/pnas.0800936105","article-title":"Extracellular loop C of NPC1L1 is important for binding to ezetimibe","volume":"105","author":"Weinglass","year":"2008","journal-title":"Proc Natl Acad Sci U S A"},{"key":"2023053006052186200_CIT0023","doi-asserted-by":"crossref","first-page":"1845","DOI":"10.1091\/mbc.e16-03-0154","article-title":"Ezetimibe-sensitive cholesterol uptake by NPC1L1 protein does not require endocytosis","volume":"27","author":"Johnson","year":"2016","journal-title":"Mol Biol Cell"},{"key":"2023053006052186200_CIT0024","doi-asserted-by":"crossref","first-page":"14824","DOI":"10.1038\/ncomms14824","article-title":"Regulation of hepatic lipogenesis by the zinc finger protein Zbtb20","volume":"8","author":"Liu","year":"2017","journal-title":"Nat Commun"},{"key":"2023053006052186200_CIT0025","doi-asserted-by":"crossref","first-page":"2023","DOI":"10.1194\/jlr.M800145-JLR200","article-title":"Kinetic imaging of NPC1L1 and sterol trafficking between plasma membrane and recycling endosomes in hepatoma cells","volume":"49","author":"Petersen","year":"2008","journal-title":"J Lipid Res"},{"key":"2023053006052186200_CIT0026","doi-asserted-by":"crossref","first-page":"609","DOI":"10.1016\/j.atherosclerosis.2014.09.036","article-title":"Genetic demonstration of intestinal NPC1L1 as a major determinant of hepatic cholesterol and blood atherogenic lipoprotein levels","volume":"237","author":"Xie","year":"2014","journal-title":"Atherosclerosis"}],"container-title":["Life Metabolism"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/academic.oup.com\/lifemeta\/advance-article-pdf\/doi\/10.1093\/lifemeta\/load011\/49543850\/load011.pdf","content-type":"application\/pdf","content-version":"am","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/lifemeta\/article-pdf\/2\/2\/load011\/50484598\/load011.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/lifemeta\/article-pdf\/2\/2\/load011\/50484598\/load011.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,5,30]],"date-time":"2023-05-30T06:05:39Z","timestamp":1685426739000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/lifemeta\/article\/doi\/10.1093\/lifemeta\/load011\/7079049"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,3,16]]},"references-count":26,"journal-issue":{"issue":"2","published-print":{"date-parts":[[2023,4,1]]}},"URL":"https:\/\/doi.org\/10.1093\/lifemeta\/load011","relation":{},"ISSN":["2755-0230"],"issn-type":[{"value":"2755-0230","type":"electronic"}],"subject":[],"published-other":{"date-parts":[[2023,4,1]]},"published":{"date-parts":[[2023,3,16]]},"article-number":"load011"}}