title: Uptake of cholesterol by the retina occurs primarily via a low density lipoprotein receptor-mediated process.
abstract: OBJECTIVE
  In this study we examined the uptake of circulating lipoproteins into the retina, using a naturally fluorescent cholesterol analog for imaging and deuterated cholesterol for quantification by mass spectroscopy. The purpose of this study was to better understand cholesterol uptake, transport and homeostasis in the retina.
  
  
  METHODS
  Human low density lipoprotein (LDL) and high density lipoprotein (HDL) were labeled with the fluorescent cholesterol analog cholesta-5,7,9(11)-trien-3beta-ol (CTL) and deuterated cholesterol (25,26,26,26,27,27,27-[2H]cholesterol, D7Ch). Rats were injected intravenously with CTL-LDL, CTL-HDL and D7Ch-LDL. Fluorescent confocal microscopy was used to image the uptake of CTL and mass spectroscopy was used to quantify D7Ch. Immunohistochemistry and fluorescent confocal microscopy were used to localize apoB (an LDL marker protein) and LDL receptor (LDLR) protein in rat and monkey retinas.
  
  
  RESULTS
  CTL-specific fluorescence was imaged by confocal microscopy in the retinal pigment epithelium (RPE), choriocapillaris and parts of the neural retina within 2 h post-injection and was visualized in the photoreceptor outer segments by 4 h. Replacing LDL with HDL as the CTL carrier gave a less robust and more delayed labeling of retinal layers. Human apolipoprotein B (apoB) was also localized in the rat choriocapillaris and RPE by 4 h post-injection. Human apoB was detected by immunoblot analysis in the rat retina primarily as a about 70 kDa protein, suggesting proteolytic degradation. LDL-mediated uptake of cholesterol was quantified by mass spectroscopy using deuterated cholesterol in place of CTL. In addition, apoB and LDLR were localized in monkey retina by immunohistochemistry.
  
  
  CONCLUSIONS
  The retina is capable of rapid uptake of circulating LDL via an LDLR-mediated process primarily occurring in the RPE and also possibly Müller cells. Despite the dominance of HDL over LDL in rat serum, LDL appears to be the preferred carrier for cholesterol transport to and uptake by the retina. The results also suggest that blood-borne LDL represents a significant contributor to the steady-state levels of cholesterol and possibly other lipids in the retina.
authors.affiliation: Laboratory of Retinal Cell and Molecular Biology, Section on Mechanisms of Retinal Diseases, National Eye Institute, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Nomingerel
authors.initials: N
authors.lastname: Tserentsoodol
authors.forename: Jorge
authors.initials: J
authors.lastname: Sztein
authors.forename: Mercedes
authors.initials: M
authors.lastname: Campos
authors.forename: Natalya V
authors.initials: NV
authors.lastname: Gordiyenko
authors.forename: Robert N
authors.initials: RN
authors.lastname: Fariss
authors.forename: Jung Wha
authors.initials: JW
authors.lastname: Lee
authors.forename: Steven J
authors.initials: SJ
authors.lastname: Fliesler
authors.forename: Ignacio R
authors.initials: IR
authors.lastname: Rodriguez
mesh_terms: D000818:Animals; D001055:Apolipoproteins B / Q000493:pharmacokinetics; D002460:Cell Line; D002782:Cholestenes / Q000493:pharmacokinetics; D002784:Cholesterol / Q000378:metabolism* / Q000493:pharmacokinetics; D006801:Humans; D007150:Immunohistochemistry; D007275:Injections, Intravenous; D008077:Lipoproteins, LDL / Q000008:administration & dosage / Q000493:pharmacokinetics; D008253:Macaca mulatta; D051381:Rats; D017207:Rats, Sprague-Dawley; D011973:Receptors, LDL / Q000378:metabolism*; D012160:Retina / Q000378:metabolism*; D014018:Tissue Distribution
chemical_list: D001055:Apolipoproteins B; D002782:Cholestenes; D008077:Lipoproteins, LDL; D011973:Receptors, LDL; C025498:cholesta-5,7,9-trien-3 beta-ol; D002784:Cholesterol