KOR

e-Article

Intracellular transport of cholesterol in mammalian cells
Document Type
Miscellaneous
Author
Source
Other Information: Thesis (Ph. D.)
Subject
59 BASIC BIOLOGICAL SCIENCES CHOLESTEROL
MEMBRANE TRANSPORT
ANTIBIOTICS
BIOASSAY
BIOLOGICAL HALF-LIFE
CELL MEMBRANES
CHO CELLS
ERYTHROCYTES
OXIDOREDUCTASES
TRACER TECHNIQUES
TRITIUM COMPOUNDS
ANIMAL CELLS
ANTI-INFECTIVE AGENTS
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BODY FLUIDS
CELL CONSTITUENTS
DRUGS
ENZYMES
HYDROGEN COMPOUNDS
HYDROXY COMPOUNDS
ISOTOPE APPLICATIONS
MATERIALS
MEMBRANES
ORGANIC COMPOUNDS
STEROIDS
STEROLS 551001* -- Physiological Systems-- Tracer Techniques
Language
English
Abstract
The erythrocyte was selected as a simple cell for the study of transbilayer movement of cholesterol. Cholesterol oxidase was used to measure the distribution of ({sup 3}H)cholesterol across the erythrocyte membrane. Cholesterol oxidase was also used to estimate the rate of transport of low density lipoprotein (LDL) cholesterol to the plasma membrane of cultured Chinese hamster ovary (CHO) fibroblasts; the half-time of this process was 42 minutes. The rate of transport of LDL cholesterol to the plasma membrane was confirmed by a second procedure using amphotericin B. Amphotericin B was also used to estimate the rate of transport of endogenously synthesized cholesterol to the plasma membrane of CHO cells. New methodology was developed including improvements of the previously published cholesterol oxidase assay for plasma membrane cholesterol. A new method for detecting transport of cholesterol to the plasma membrane in cultured cells was developed using amphotericin B. Preliminary studies investigated the use of fluorescent polyenes, pimaricin and etruscomycin, as probes for plasma membrane cholesterol in transport studies. Finally, a modification of a previously published cell staining protocol yielded a simple, quantitative assay for cell growth.