title: Quantitative and qualitative methods using fluorescence microscopy for the study of modified low density lipoproteins uptake.
abstract: Atherosclerosis and heart disease are the main cause of death in United States. The development of atherosclerosis includes lipid deposition and foam cell formation in the artery wall. Scavenger Receptors A-I and II (SRA-I/II) have an important role of in foam cell formation and atherogenesis. Most of the SRA-I/II studies had been performed using Iodine-125-radiolabeled modified low-density lipoprotein. This report attempts to validate the use of fluorescence microscopy techniques as an alternative to obtain qualitative and quantitative information of the uptake of fluorescence-labeled acetylated low-density lipoprotein (AcLDL) in adherent CHO cells expressing SRA-I/II. After verifying the protein expression of SRA-I and II, uptake was quantified using a Laser Scan Cytometer, and images of cells containing fluorescent AcLDL were obtained. A significant increase in fluorescence was found in the cells transfected with SRA-I/II vs. those with empty vector. When SRA-I/II competitive ligands were used, the uptake of AcLDL was significantly decreased. In conclusion, the use of fluorescence microscopy techniques in obtaining qualitative and quantitative information of the uptake of fluorescence-labeled AcLDL by adherent cells, such as CHO cells, is an alternative to the traditional use of radiolabeled iodine.
authors.affiliation: Division of Lung Diseases, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland, USA. leyvaf@mail.nih.gov
authors.forename: Francisco J
authors.initials: FJ
authors.lastname: Leyva
authors.forename: Mark A
authors.initials: MA
authors.lastname: Pershouse
mesh_terms: D000107:Acetylation; D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D005456:Fluorescent Dyes; D008077:Lipoproteins, LDL / Q000378:metabolism*; D051379:Mice; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence / Q000379:methods*; D051116:Receptors, Scavenger / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
chemical_list: D005456:Fluorescent Dyes; D008077:Lipoproteins, LDL; D051116:Receptors, Scavenger