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(Received for publication, April 3, 1997, and in revised form, April 21, 1997)
From the ABSTRACT Lecithin:cholesterol acyltransferase (LCAT) is the
major determinant of the cholesteryl ester (CE) content of high density lipoprotein (HDL) in plasma. The selective uptake of HDL-CE is postulated to participate in delivery of tissue-derived cholesterol both to the liver and steroidogenic tissues. Recent studies comparing mice with similarly low levels of HDL, due to the absence of either of
the two major HDL-associated apolipoproteins apoA-I and apoA-II, suggest that apoA-I is crucial in modulating this process, possibly through interaction with scavenger receptor class B type I (SR-BI). Because of the central role of LCAT in determining the size, lipid composition, and plasma concentration of HDL, we have created LCAT-deficient mice by gene targeting to examine the effect of LCAT
deficiency on HDL structure and composition and adrenal cholesterol delivery. The HDL in the LCAT-deficient mice was reduced in its plasma
concentration (92%) and CE content (96%). The HDL particles were
heterogeneous in size and morphology and included numerous discoidal
particles, mimicking those observed in LCAT-deficient humans. The
adrenals of the male Lcat ( INTRODUCTION High density lipoprotein (HDL)1 plays a
pivotal role in lipid homeostasis by removing cholesterol accumulating
in the extrahepatic tissues, a process crucial for maintenance of the
structure and function of most cells in the body. The mechanism widely
held to explain this process, reverse cholesterol transport (1), describes the metabolic fate of such tissue-derived cholesterol. According to this mechanism, circulating HDL functions as an acceptor of tissue-derived unesterified cholesterol (UC), facilitating efflux of
cholesterol from cells. Lecithin:cholesterol acyltransferase (LCAT)
performs a central role in this process by catalyzing the conversion of
plasma UC, especially that associated with HDL, to cholesteryl ester
(CE). It has been demonstrated that the majority of the CE in HDL is
delivered to the liver for clearance or recycling. In addition, a
fraction of HDL-CE is delivered to the steroidogenic tissues and is
utilized as substrate for steroid hormone synthesis. Routes for
delivery of HDL-CE to the target organs include: (i) indirectly, by
transferring to apolipoprotein (apo) B-containing lipoprotein
particles, (ii) directly through a receptor-mediated endocytotic
process (2, 3), and (iii) via selective uptake pathway where CE in
lipoprotein particles is delivered into the cells without simultaneous
uptake of the apolipoprotein moieties (4-6). Although numerous
patients with a complete deficiency of LCAT have been described,
revealing many of the lipoprotein abnormalities and tissue organ damage
associated with complete absence of LCAT, there remain many unanswered
questions concerning the role of LCAT in the transport of CE to
tissues.
To date, much of our knowledge of the selective uptake pathway derives
from studies on rodents. In rats, not only is the selective uptake
pathway responsible for 65% of HDL-CE delivered to the liver, it is
the principal mechanism for delivery of CE to the steroidogenic tissues
(4). Murine scavenger receptor class B type I (SR-BI), recently
characterized as a potential HDL receptor (7), is believed to
participate in mediating the selective uptake of HDL-CE into cells. Key
evidence supporting this role for SR-BI includes the tissue expression
pattern of SR-BI being coincident with those involved in selective
uptake and the observation that SR-BI expression in the adrenal is
strongly regulated by intracellular cholesterol content (8). Recently,
an in vivo study comparing apoA-I and apoA-II gene-targeted
mice, both with similarly low HDL levels, demonstrated that only the
lack of apoA-I severely impairs the delivery of CE to the steroidogenic
tissues, causing depletion in adrenal CE stores and blunted
steroidogenic responses to stress (9). Although the different
consequences of the two low HDL states strongly suggest the crucial
importance of apoA-I in HDL in mediating selective uptake of CE, the
question of whether there are additional independent modulating factors remains unanswered.
In this study, we tested the hypothesis that HDL resulting from LCAT
deficiency, being severely depleted of CE yet containing apoA-I,
impairs cholesterol delivery to the adrenals. To investigate this
issue, LCAT-deficient mice were created by gene targeting. The effect
of complete LCAT deficiency on lipoproteins and cholesterol transport
was characterized in these animals.
MATERIALS AND METHODS The strategy for
construction of the targeting vector pPN2T/LKO is shown in Fig. 1. The
targeting vector was linearized and transfected into ES cells by
electroporation. Drug selection was performed using both G418 (160 µg/ml) and FIAU (0.5 µM). Genomic Southern blot was
used for screening of targeting events using a PCR based probe that
hybridizes to exon 6 of the Lcat gene. The targeted ES cells were
injected into blastocysts by standard method (10). Chimeric mice were
bred with DBAxC57BL/6 F1 hybrids. Genotypes were determined by
multiplex PCR. The forward primers either hybridize specifically to the
neo-resistant gene (a, 5
LCAT activity was measured on
fasting plasma as the rate of synthesis of
[3H]cholesteryl esters from unilamellar vesicles prepared
with French pressure cell and activated with human apoA-I (Sigma) as
described previously (11).
Lipid determinations
were performed on 4-8-week-old mice fed a mouse chow diet. Fasting
plasma was analyzed using enzyme end point kits for total cholesterol
(Boehringer Mannheim), free glycerol, and triglyceride concentrations
(Sigma). Plasma triglyceride concentration was corrected for free
glycerol content. Cholesteryl ester in lipoprotein fractions was
calculated from difference between total and unesterified cholesterol;
HDL-cholesterol was measured in plasma after selective precipitation of
apoB-containing lipoproteins by polyethylene glycol. Mouse plasma
apoA-I levels were quantified by enzyme-linked immunosorbent assay as
described previously (12).
The relative proportion of HDL with pre- Tissue lipids
were measured in the kidney, liver, spleen, heart, lung, skeletal
muscle, and adrenal gland of both the Lcat ( Mice were perfused
through the left ventricle with phosphate-buffered saline followed by
overnight fixation in 4% formaldehyde. The adrenal glands were
infiltrated sequentially with 10, 20, and 30% sucrose in
phosphate-buffered saline and quick frozen in Tissue-Tec OCT.
6-10-µm-thick sections were placed on poly-L-lysine coated slides, stained with Oil Red O, and counterstained with hematoxylin (18).
RNA was isolated from single adrenals from five
male Lcat ( RESULTS Genomic Southern blot
screening of 200 ES cells targeted with pPN2T/LKO yielded a frequency
of 1 in 4 for correct homologous recombination event. 2 of the 10 targeted ES cell clones micro-injected gave germ-line transmission of
the targeted allele.
Multiplex PCR screening of the targeted mutant is shown in Fig.
1B. Positivity from primer sets b/c alone, a/c
plus b/c, and a/c alone identified wild type, heterozygosity, and
homozygosity for the targeted Lcat allele respectively. Of 86 offspring
from the heterozygote breeding, 25 were wild type, 39 were
heterozygotes, and 22 were homozygotes (p = 0.73 by chi
square test against the expected 1:2:1 Mendelian inheritance). The
LCAT-deficient mice grow normally and are fertile.
Plasma LCAT activity of the mutant mice is shown in Fig. 1C.
The Lcat ( Compared with wild type
controls, lipid analyses on the Lcat ( Table I.
Plasma lipids, lipoproteins, and apoA-I levels in Lcat(+/+),
Lcat(+/ Table II.
Lipid analyses of plasma subfractions in Lcat(+/+) and Lcat( Plasma were pooled from five mice in each group; density subfractions
were obtained by sequential
ultracentrifugation.
Volume 272, Number 25,
Issue of June 20, 1997
pp. 15777-15781
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.
§,
,,**
Human Genome Center, Life Sciences Division,
Lawrence Berkeley National Laboratory, University of California,
Berkeley, California 94720, ¶ Pfizer Inc., the Department of
Cardiovascular and Metabolic Diseases, Central Research Division,
Groton, Connecticut 06340, and the 
Donner Laboratory, University
of California and Molecular Medicine Research Program, Lawrence
Berkeley National Laboratory, University of California,
Berkeley, California 94720
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
FOOTNOTES
ACKNOWLEDGEMENTS
REFERENCES
/) mice were severely depleted of lipid stores, which was associated with a 2-fold
up-regulation of the adrenal SR-BI mRNA. These studies demonstrate
that LCAT deficiency, similar to apoA-I deficiency, is associated
with a marked decrease in adrenal cholesterol delivery and supports the hypothesis that adrenal SR-BI expression is regulated by the adrenal cholesterol.
-AAGCAAAACCAAATTAAGGGC-3) or to the targeted
region (b, 5-GCTCCTCAATGTGCCTTCC-3) and share a common reverse primer
which hybridizes to exon 3 of the gene (c,
5-GTACTCAACAGATTCGGTCTTGC-3) (Fig. 1A).
Fig. 1.
Generation of Lcat (/) mice.
A, targeting vector design and strategy. Exons are indicated
by raised boxes and are numbered. Shaded lines
represent the long and short arms of sequence homology used in the
targeting vector, pPN2T/LKO. The unique NotI (N)
site in pPN2T/LKO was used to linearize the vector. Targeted ES clones
were identified by a unique 5.8-kilobase (kb)
HindIII fragment as detected by a 0.5-kilobase PCR-generated
probe that hybridizes to exon 6. Black arrows represent
locations of primers used for multiplex PCR genotyping. PCR was
performed on mouse tail genomic DNA using primers a, b, and c.
X, XbaI; H, HindIII; S, Sse8387I; B, BamHI.
B, multiplex PCR identification of mouse genotypes.
WT, wild type. C, LCAT activity of mouse plasma
using the exogenous substrate assay. The bar graph
represents the means ± S.D. from 13 Lcat (+/+) and 9 Lcat (+/) animals. ND, nondetectable. *,
p < 0.001 for Lcat (+/) versus
Lcat (+/+).
[View Larger Version of this Image (36K GIF file)]
- and
-mobility on agarose gel in the Lcat (+/+) and
Lcat (/) mice was determined as described previously
(13). Lipoprotein subfractions of d < 1.063 and
d = 1.063-1.21 g/ml were obtained from a pool of
200-250 µl of plasma by standard sequential ultracentrifugation
techniques. The size distribution of the lipoprotein fractions were
obtained by nondenaturing polyacrylamide gradient gel electrophoresis
essentially as described by Nichols et al. (14). The
morphology of lipoproteins was assessed by negative staining electron
microscopy as described previously (15).
/) and
Lcat (+/+) mice. After perfusion of the animal with 0.9%
saline, the organs were excised, blotted dry, weighed, and homogenized
in 1.15% KCl buffer. Lipids were extracted using the Bligh and Dyer
method (16) and reconstituted in the aqueous phase using Triton X-100.
Total cholesterol and UC were measured using an enzymatic assay
(17).
/) and five male wild type mice using RNA
STAT-60 kit (Tel-Test "B", Inc.) and separated in a denaturing 1%
agarose gel containing 2.2 M formaldehyde. RNA was
transferred to Nylon membranes (Schleider & Schuell), cross-linked, and
hybridized with a 554-base pair PCR fragment extending from nucleotides
362 to 915 of the murine SR-BI. The SR-BI mRNA Northern blot signal
was normalized to that of the human -actin probe
(CLONTECH Laboratories, Inc., CA) on the same
membrane.
/) mice had no detectable LCAT activity,
whereas the heterozygous mice had LCAT activity reduced to 55% of
control (p = 0.0001).
/) mice revealed
70.3 and 91.6% reductions in total and HDL cholesterol, respectively
(Table I). Plasma CE was reduced to 9.4% of control,
whereas plasma UC level was essentially unchanged. As a result, there
was a 11.7-fold increase in the UC/CE ratio compared with the wild type
littermates. The Lcat (+/) mice experienced a more
moderate but statistically significant reduction in total cholesterol
and CE, resulting in a modest 1.7-fold increase in the UC/CE ratio. The
genotype-dependent changes in UC/CE ratio was nearly
entirely due to changes in plasma CE levels. Likewise, the reduction in
total cholesterol in the Lcat (/) mice was largely a
result of reduction in HDL cholesterol levels, whereas the non-HDL
cholesterol levels for each genotype were essentially unchanged. Plasma
apoA-I levels in the mutant mice were also reduced in both
Lcat (/) and Lcat (+/) mice to 19 and 86%
of their respective controls. Lipid analyses on d < 1.063 and d = 1.063-1.21 g/ml fractions from pooled
plasma are shown in Table II. The reduction of HDL-CE in
the Lcat (/) mice was profound, whereas the HDL-UC reduction in the same mice was less dramatic. Agarose gel
electrophoresis of HDL showed that compared with wild type control, the
fraction of plasma apoA-I in the -migrating HDL particles in the
Lcat (/) mice was decreased from 93.6 ± 0.8% to
78.3 ± 3.5% (p < 0.01) and was increased from
6.4 ± 0.8% to 21.7 ± 3.5% (p < 0.01) in
the pre--migrating HDL.
), and Lcat(/) mice
Genotype
T.
Chol.
TG
UC
HDL-C
UC/CE
apoA-I
mg/dl (n = 7)
mg/dl
+/+
111 ± 22
67
± 50
25 ± 8
84 ± 19
0.23 ± 0.06
184.7
± 30.5 (n = 6)
+/
69 ± 19a
78
± 42
21 ± 7
50 ± 16a
0.40
± 0.17b
160.5
± 21.1 (n = 8)
/33
± 12a
117 ± 88
24 ± 11
7
± 3a
2.71 ± 1.11b
34.9
± 9.4 (n = 5)a
a
p < 0.00003 compared with wild type
control.
b
p < 0.002 compared with wild type control.
/)
mice
Genotype
d < 1.063 g/ml
d = 1.063-1.21 g/ml
T.
Chol.
UC
CE
T.
Chol.
UC
CE
mg/dl
+/+
15.6
10.6
5
90.4
22.1
68.3
/19.5
14.5
5
16.0
13.4
2.6
The lipoprotein
size distribution of the Lcat (+/+) and Lcat
(/) mice in d = 1.063-1.21 g/ml fractions are
shown in Fig. 2. The pattern of the d < 1.063 g/ml fraction of Lcat (/) mice, in comparison with
that of the wild type, showed absence of the 20.5 nm peak, broadening
of the 28.0 nm LDL peak to include intermediate density lipoprotein in
the region of 32.1 nm, and preservation of the dominant VLDL peak.
Unlike a unimodal size distribution of HDL in the wild types, the
Lcat (/) mice had a very complex pattern, characterized
by a major peak at 7.6 nm and several minor components similar to that
found in human LCAT deficiency (19). Electron microscopic evaluation of
VLDL from the control mice revealed spherical particles with a mean
diameter (d) and a S.D. of 63.5 ± 19.1 nm, whereas
VLDL from Lcat (/) mice were smaller (d ± S.D. = 51.7 ± 22.8 nm) and were distinguished by the presence of notched particles (Fig. 3). The HDL from Lcat
(/) mice, compared with control HDL that were homogeneous spherical
particles of 10.0 ± 2.1 nm in diameter, were morphologically
complex. An important observation, however, is the presence of
discoidal particles that form the classical rouleaux structures. Both
morphologic features have been described in LCAT-deficient subjects
(19, 20). Larger particles were also present and may represent
contamination from the d < 1.063 g/ml fraction.
Fig. 2. Nondenaturing gradient gel electrophoresis of plasma lipoprotein subfractions pooled from Lcat (+/+) (dashed line) and Lcat (
/) (solid
line) mice. The listed peak size is in nanometers. A, the d < 1.063 g/ml fraction was analyzed
on 2-16% gels and stained with Oil Red O. B, the
d = 1.063-1.21 g/ml fraction was analyzed on 4-30%
gels and stained with Coomassie G250.
[View Larger Version of this Image (17K GIF file)]
Fig. 3. Electron micrographs of negatively stained plasma lipoprotein fractions of Lcat (+/+) and Lcat (
/) mice. A, VLDL fraction,
Lcat (+/+) mice; B, VLDL fraction,
Lcat (/) mice; C, HDL fraction,
Lcat (+/+) mice; and D, HDL fraction,
Lcat (/) mice. The large sized particles in the
Lcat (/) HDL fraction represent contaminating VLDL/LDL
particles. The arrow in B indicates a notched
VLDL particle. The bar markers represent 100 nm.
[View Larger Version of this Image (132K GIF file)]
Analysis of the Effect of LCAT Deficiency on the Target Tissues
For the total cholesterol and UC in the nonsteroidogenic
tissues examined, no significant difference between the Lcat
(/) and Lcat (+/+) mice was noted, with the exception of
the spleen, which showed a 21% increase in UC content
(p = 0.047) but not in total cholesterol (data not
shown). After perfusion, the adrenal glands of the Lcat
(/) mice appeared brown and translucent, in contrast to a pearly
white appearance of adrenals from the wild type mice (Fig.
4A), consistent with the depletion of cholesterol in the former. Staining of the adrenal sections with Oil Red O demonstrated an abundance of stored lipid in the adrenal cortex of the
wild type mice. In contrast, the Lcat (/) mice showed complete absence of lipid staining (Fig. 4B). Quantification
of tissue lipid in the adrenal glands showed a 61% reduction in total tissue cholesterol, an 82% reduction in CE content, and a 30% reduction in UC content in the Lcat (/) mice compared
with the wild type control (Fig. 4C). The normalized adrenal
SR-BI mRNA expression levels was 2.0-fold higher in the
Lcat (/) mice compared with the wild type control
(p = 0.001) (Fig. 4D).
Fig. 4. Adrenal histology, lipid content and SR-BI mRNA levels in Lcat (+/+) and Lcat (
/)
mice. A, gross appearance of adrenal glands from male
Lcat (+/+) and Lcat (/) mice. The scale shown is in millimeters. B, Oil Red O stained frozen sections of
the adrenals showing complete depletion of neutral lipid store in the
adrenal cortex of male Lcat (/). C,
quantification of tissue cholesterol in the adrenals of Lcat
(+/+) and Lcat (/) mice. Bar graph represents
the means ± S.D. from three animals in each group. *,
p = 0.008, and **, p = 0.09, compared
with their respective wild type controls. D, quantification
of mouse SR-BI mRNA expression by Northern blot in Lcat
(+/+) and Lcat (/) mice. PhosphorImager quantification
of the SR-BI mRNA bands after normalization to the -actin level.
The bar graph represents the means ± S.D. from five
animals in each group. *, p < 0.05 compared with wild
type control.
[View Larger Version of this Image (67K GIF file)]
DISCUSSION
A complete absence of LCAT activity was observed in the
Lcat (/) mice, whereas the LCAT activity of the
Lcat (+/) mice was reduced to approximately half normal,
compatible with autosomal co-dominant inheritance. The same inheritance
has been reported in a human LCAT-deficient kindred (21). The
alterations of HDL in the Lcat (/) mice also resembled
those observed in LCAT-deficient humans. This includes a comparable
degree of reduction in both plasma HDL cholesterol and apoA-I and the
presence of unique discoidal particles with rouleaux formation in the
Lcat (/) mice. Furthermore, we observed a higher
proportion of the HDL particles in the Lcat (/) mice
with pre- mobility, a finding also noted in LCAT-deficient patients.
The impact of LCAT deficiency on non-HDL cholesterol is characterized by a modest increase in the UC level. However, gradient gel electrophoresis and electron microscopy studies showed presence of intermediate density lipoprotein-like particles and notching of VLDL, respectively, in the LCAT-deficient mice. In view of the absence of cholesteryl ester transfer activity in mouse plasma, these findings suggest that LCAT may have a direct effect on the metabolism of the triglyceride-rich lipoproteins (22).
Despite its central role in reverse cholesterol transport, the impact
of LCAT deficiency on plasma cholesterol transport and delivery to
target organs has been minimally explored. In LCAT-deficient humans,
the rate of cholesterol efflux into plasma is preserved at 72% of
normal. This is in part explained by the relative abundance of
pre--HDL particles, a subpopulation of HDL believed to be the
preferred initial acceptors of tissue cholesterol (23). The relative
increase in pre--HDL is also observed in the LCAT-deficient mice.
However, it has not been determined whether the extremely low level of
abnormal HDL associated with LCAT deficiency is able to deliver
tissue-derived CE to the target organs. The recent study by Plump
et al. (9) demonstrating the crucial role that apoA-I plays
in the delivery of CE through the selective uptake pathway and the
observation by Wang et al. (8) that murine adrenal SR-BI
receptor expression being up-regulated in association with
intracellular cholesterol depletion suggest that ligand-receptor interaction between apoA-I and SR-BI may be a major determinant of
effective mediation of CE delivery into the tissues. Our similar observation of severe depletion of tissue lipid stores in mouse adrenals and the finding of a 2-fold up-regulation of adrenal SR-BI
mRNA in face of adrenal cholesterol depletion in Lcat
(/) mice suggests that (i) SR-BI is intimately involved in
selective uptake of CE and (ii) this same metabolic pathway is
defective in the Lcat (/) mice despite the presence of
apoA-I in HDL. As in apoA-I-deficient mice, it can be inferred that the
minor sources of adrenal tissue cholesterol, namely de novo
synthesis and LDL receptor mediated cholesterol uptake, are
insufficient to compensate for the cholesterol depletion. In
Lcat (/) mice, the most likely cause for reduced adrenal
lipid content is the severe depletion of plasma HDL-CE, the primary
source of cellular cholesterol.
The impact of LCAT deficiency on the other tissues examined was
significantly less than that observed for the adrenal. A hallmark finding in LCAT-deficient subjects is the marked tissue UC accumulation in a number of solid organs including the liver, spleen, and kidney. With the exception of a slight increase in UC in the spleen, these lipid disturbances were not observed in the nonsteroidogenic tissues in
the Lcat (/) mice. The mild phenotype in these mice is
likely due to the relatively small reservoir of total plasma UC level with the consequent minor impact on tissue lipid homeostasis (24).
In summary, the LCAT-deficient mouse created by gene targeting reproduces the human LCAT-deficient HDL metabolic abnormalities with high fidelity. The noted morphologic abnormalities of VLDL suggest that LCAT may have a direct role in the catabolism of triglyceride-rich, apoB-containing particles. Finally, these studies suggest that the reduced cholesterol content of HDL from deficiency of LCAT likely impacts on the flux of cholesterol to the adrenals via the selective uptake pathway.
FOOTNOTES
§ Supported in part by a Heart and Stroke Foundation of Canada Research Fellowship and a Medical Research Council of Canada Research Fellowship.
** American Heart Association Established Investigator. To whom correspondence should be addressed: Human Genome Center, M/S 74-157, Lawrence Berkeley National Laboratory, University of California, Berkeley, CA 94720. Tel.: 510-486-5072; Fax: 510-486-6746; E-mail: emrubin{at}lbl.gov.
1 The abbreviations used are: HDL, high density lipoprotein(s); LCAT, lecithin:cholesterol acyltransferase; apo, apolipoprotein; SR-BI, scavenger receptor class B type I; CE, cholesteryl ester; UC, unesterified cholesterol; PCR, polymerase chain reaction; LDL, low density lipoprotein(s); VLDL, very low density lipoprotein(s).
ACKNOWLEDGEMENTS
We are indebted to Laura Knoff, Judy Verstuyft, Pat Blanche, Laura Holl, and the Donner Core Lipoprotein Analysis laboratory for technical support; Elaine Gong for critical reading of the manuscript; and Drs. Chris Paszty, John Bielicki, and John Morrison for valuable discussions.
REFERENCES
- Glomset, J. A. (1968) J. Lipid Res. 9, 155-167 [Abstract]
-
Chao, Y., Windler, E. E., Chen, G. C., and Havel, G. C.
(1979)
J. Biol. Chem.
254,
11360-11366
[Free Full Text] - Drevon, C. A., Attie, A. D., Pangburn, S. H., and Steinberg, D. (1981) J. Lipid Res. 22, 37-46 [Abstract]
-
Glass, C., Pittman, R. C., Civen, M., and Steinberg, D.
(1985)
J. Biol. Chem.
260,
744-750
[Abstract/Free Full Text] -
Pittman, R. C., Knecht, T. P., Rosenbaum, M. S., and Taylor, C. A., Jr.
(1987)
J. Biol. Chem.
262,
2443-2450
[Abstract/Free Full Text] -
Glass, C., Pittman, R. C., Weinstein, D. B., and Steinberg, D.
(1983)
Proc. Natl. Acad. Sci. U. S. A.
80,
5435-5439
[Abstract/Free Full Text] - Acton, S., Rigotti, A., Landschulz, K. T., Xu, S., Hobbs, H. H., and Krieger, M. (1996) Science 271, 518-520 [Abstract]
-
Wang, N., Weng, W., Breslow, J., and Tall, A. R.
(1996)
J. Biol. Chem.
271,
21001-21004
[Abstract/Free Full Text] - Plump, A. S., Erickson, S. K., Weng, W., Partin, J. S., Breslow, J. L., and Williams, D. L. (1996) J. Clin. Invest. 97, 2660-2671 [Medline] [Order article via Infotrieve]
- Paszty, C., Narla, M., Stevens, M., Liebhaber, S. A., Loring, J. F., Brion, C. M., and Rubin, E. M. (1995) Nat. Genet. 11, 33-39 [CrossRef][Medline] [Order article via Infotrieve]
- Francone, O. L., Gong, E. L., Ng, D. S., Fielding, C. J., and Rubin, E. M. (1995) J. Clin. Invest. 96, 1440-1448
- Francone, O. L., Haghpassand, M., Benett, J. A., Royer, L., and McNeish, J. (1996) J. Lipid Res. 38, 813-822 [Abstract]
- Francone, O. L., Royer, L., and Haghpassand, M. (1996) J. Lipid Res. 37, 1268-1277 [Abstract]
- Nichols, A. R., Krauss, R. M., and Muslliner, T. (1986) Methods Enzymol. 128, 417-431 [Medline] [Order article via Infotrieve]
- Forte, T. M., and Nordhausen, R. W. (1986) Methods Enzymol. 128, 442-457 [Medline] [Order article via Infotrieve]
- Bligh, E. G., and Dyer, W. J. (1959) Can. J. Biochem. 37, 911-917 [Medline] [Order article via Infotrieve]
- Sale, F. O., Marchesini, S., Fishman, P. H., and Berra, B. (1984) Anal. Biochem. 142, 347-350 [CrossRef][Medline] [Order article via Infotrieve]
- Coalson, R. E. (1981) in Staining Procedures (Clark, G., ed), pp. 228-229, Williams and Wilkins, Baltimore, MD
- Mitchell, C. D., King, W. C., Applegate, K. R., Forte, T., Glomset, J. A., Norum, K. R., and Gjone, E. (1980) J. Lipid Res. 21, 625-634 [Abstract]
- Forte, T. M., Nichols, A., Glomset, J., and Norum, K. (1974) Scand. J. Clin. Lab. Invest. 33Suppl. 137 (Suppl. 137), 121-132 [Medline] [Order article via Infotrieve]
- Jain, S. K., Mohandas, N., Sensabaugh, G. F., Shojania, A. M., and Shohet, S. B. (1982) J. Lab. Clin. Med. 99, 816-826 [Medline] [Order article via Infotrieve]
- Schumaker, V. N., and Adams, G. H. (1970) J. Theor. Biol. 26, 89-91 [CrossRef][Medline] [Order article via Infotrieve]
-
von Ackerstein, A., Huang, Y., Wu, S., Funke, H., Noseda, G., and Assmann, G.
(1995)
Arterioscler. Thromb. Vasc. Biol.
15,
691-703
[Abstract/Free Full Text] - Hovig, T., and Gjone, E. (1974) Scand. J. Clin. Lab. Invest. 33Suppl. 137 (Suppl. 137), 135-146
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 A. Yesilaltay, M. G. Morales, L. Amigo, S. Zanlungo, A. Rigotti, S. L. Karackattu, M. H. Donahee, K. F. Kozarsky, and M. Krieger Effects of Hepatic Expression of the High-Density Lipoprotein Receptor SR-BI on Lipoprotein Metabolism and Female Fertility Endocrinology, April 1, 2006; 147(4): 1577 - 1588. [Abstract] [Full Text] [PDF]
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H. Song, L. Zhu, C. M. Picardo, G. Maguire, V. Leung, P. W. Connelly, and D. S. Ng Coordinated alteration of hepatic gene expression in fatty acid and triglyceride synthesis in LCAT-null mice is associated with altered PUFA metabolism Am J Physiol Endocrinol Metab, January 1, 2006; 290(1): E17 - E25. [Abstract] [Full Text] [PDF]
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 K. Namekata, Y. Enokido, I. Ishii, Y. Nagai, T. Harada, and H. Kimura Abnormal Lipid Metabolism in Cystathionine {beta}-Synthase-deficient Mice, an Animal Model for Hyperhomocysteinemia J. Biol. Chem., December 17, 2004; 279(51): 52961 - 52969. [Abstract] [Full Text] [PDF]
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 X. Zhu, A. M. Herzenberg, M. Eskandarian, G. F. Maguire, J. W. Scholey, P. W. Connelly, and D. S. Ng A Novel in Vivo Lecithin-Cholesterol Acyltransferase (LCAT)-Deficient Mouse Expressing Predominantly LpX Is Associated with Spontaneous Glomerulopathy Am. J. Pathol., October 1, 2004; 165(4): 1269 - 1278. [Abstract] [Full Text] [PDF]
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D. S. Ng, C. Xie, G. F. Maguire, X. Zhu, F. Ugwu, E. Lam, and P. W. Connelly Hypertriglyceridemia in Lecithin-cholesterol Acyltransferase-deficient Mice Is Associated with Hepatic Overproduction of Triglycerides, Increased Lipogenesis, and Improved Glucose Tolerance J. Biol. Chem., February 27, 2004; 279(9): 7636 - 7642. [Abstract] [Full Text] [PDF]
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 A. Braun, S. Zhang, H. E. Miettinen, S. Ebrahim, T. M. Holm, E. Vasile, M. J. Post, D. M. Yoerger, M. H. Picard, J. L. Krieger, et al. Probucol prevents early coronary heart disease and death in the high-density lipoprotein receptor SR-BI/apolipoprotein E double knockout mouse PNAS, June 10, 2003; 100(12): 7283 - 7288. [Abstract] [Full Text] [PDF]
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 A. Rigotti, H. E. Miettinen, and M. Krieger The Role of the High-Density Lipoprotein Receptor SR-BI in the Lipid Metabolism of Endocrine and Other Tissues Endocr. Rev., June 1, 2003; 24(3): 357 - 387. [Abstract] [Full Text] [PDF]
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J. Julve, J. C. Escola-Gil, V. Ribas, F. Gonzalez-Sastre, J. Ordonez-Llanos, J. L. Sanchez-Quesada, and F. Blanco-Vaca Mechanisms of HDL deficiency in mice overexpressing human apoA-II J. Lipid Res., October 1, 2002; 43(10): 1734 - 1742. [Abstract] [Full Text] [PDF]
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 C.-a. Wu, M. Tsujita, K. Okumura-Noji, S. Usui, H. Kakuuchi, M. Okazaki, and S. Yokoyama Cholesteryl Ester Transfer Protein Expressed in Lecithin Cholesterol Acyltransferase-Deficient Mice Arterioscler. Thromb. Vasc. Biol., August 1, 2002; 22(8): 1347 - 1353. [Abstract] [Full Text] [PDF]
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H. A. Feister, B. J. Auerbach, L. A. Cole, B. R. Krause, and S. K. Karathanasis Identification of an IL-6 response element in the human LCAT promoter J. Lipid Res., June 1, 2002; 43(6): 960 - 970. [Abstract] [Full Text] [PDF]
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J. A. Epstein, D. J. Rader, and M. S. Parmacek Perspective: Cardiovascular Disease in the Postgenomic Era--Lessons Learned and Challenges Ahead Endocrinology, June 1, 2002; 143(6): 2045 - 2050. [Full Text] [PDF]
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 W. A. Pitman, R. Korstanje, G. A. Churchill, E. Nicodeme, J. J. Albers, M. C. Cheung, M. A. Staton, S. S. Sampson, S. Harris, and B. Paigen Quantitative trait locus mapping of genes that regulate HDL cholesterol in SM/J and NZB/B1NJ inbred mice Physiol Genomics, May 10, 2002; 9(2): 93 - 102. [Abstract] [Full Text] [PDF]
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D. S. Ng, G. F. Maguire, J. Wylie, A. Ravandi, W. Xuan, Z. Ahmed, M. Eskandarian, A. Kuksis, and P. W. Connelly Oxidative Stress Is Markedly Elevated in Lecithin:Cholesterol Acyltransferase-deficient Mice and Is Paradoxically Reversed in the Apolipoprotein E Knockout Background in Association with a Reduction in Atherosclerosis J. Biol. Chem., March 29, 2002; 277(14): 11715 - 11720. [Abstract] [Full Text] [PDF]
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J. W. Furbee Jr., O. Francone, and J. S. Parks In vivo contribution of LCAT to apolipoprotein B lipoprotein cholesteryl esters in LDL receptor and apolipoprotein E knockout mice J. Lipid Res., March 1, 2002; 43(3): 428 - 437. [Abstract] [Full Text] [PDF]
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J. W. Furbee Jr., J. K. Sawyer, and J. S. Parks Lecithin:Cholesterol Acyltransferase Deficiency Increases Atherosclerosis in the Low Density Lipoprotein Receptor and Apolipoprotein E Knockout Mice J. Biol. Chem., January 25, 2002; 277(5): 3511 - 3519. [Abstract] [Full Text] [PDF]
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S. Tomimoto, M. Tsujita, M. Okazaki, S. Usui, T. Tada, T. Fukutomi, S. Ito, M. Itoh, and S. Yokoyama Effect of Probucol in Lecithin-Cholesterol Acyltransferase-Deficient Mice : Inhibition of 2 Independent Cellular Cholesterol-Releasing Pathways In Vivo Arterioscler. Thromb. Vasc. Biol., March 1, 2001; 21(3): 394 - 400. [Abstract] [Full Text] [PDF]
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J. C. Escolà-Gil, J. Julve, A. Marzal-Casacuberta, J. Ordóñez-Llanos, F. González-Sastre, and F. Blanco-Vaca ApoA-II expression in CETP transgenic mice increases VLDL production and impairs VLDL clearance J. Lipid Res., February 1, 2001; 42(2): 241 - 248. [Abstract] [Full Text]
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M. Mehrabian, L. W. Castellani, P.-Z. Wen, J. Wong, T. Rithaporn, S. Y. Hama, G. P. Hough, D. Johnson, J. J. Albers, G. A. Mottino, et al. Genetic control of HDL levels and composition in an interspecific mouse cross (CAST/Ei C57BL/6J) J. Lipid Res., December 1, 2000; 41(12): 1936 - 1946. [Abstract] [Full Text]
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J. C. Escolà-Gil, J. Julve, A. Marzal-Casacuberta, J. Ordóñez-Llanos, F. González-Sastre, and F. Blanco-Vaca Expression of human apolipoprotein A-II in apolipoprotein E-deficient mice induces features of familial combined hyperlipidemia J. Lipid Res., August 1, 2000; 41(8): 1328 - 1338. [Abstract] [Full Text]
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V. Terpstra, E. S. van Amersfoort, A. G. van Velzen, J. Kuiper, and T. J. C. van Berkel Hepatic and Extrahepatic Scavenger Receptors : Function in Relation to Disease Arterioscler. Thromb. Vasc. Biol., August 1, 2000; 20(8): 1860 - 1872. [Full Text] [PDF]
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J. S. Parks, K. W. Huggins, A. K. Gebre, and E. R. Burleson Phosphatidylcholine fluidity and structure affect lecithin:cholesterol acyltransferase activity J. Lipid Res., April 1, 2000; 41(4): 546 - 553. [Abstract] [Full Text]
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D. Lopez and M. P. McLean Sterol Regulatory Element-Binding Protein-1a Binds to cis Elements in the Promoter of the Rat High Density Lipoprotein Receptor SR-BI Gene Endocrinology, December 1, 1999; 140(12): 5669 - 5681. [Abstract] [Full Text]
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Y. Sun, N. Wang, and A. R. Tall Regulation of adrenal scavenger receptor-BI expression by ACTH and cellular cholesterol pools J. Lipid Res., October 1, 1999; 40(10): 1799 - 1805. [Abstract] [Full Text]
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 G. Cao, L. Zhao, H. Stangl, T. Hasegawa, J. A. Richardson, K. L. Parker, and H. H. Hobbs Developmental and Hormonal Regulation of Murine Scavenger Receptor, Class B, Type 1 Mol. Endocrinol., September 1, 1999; 13(9): 1460 - 1473. [Abstract] [Full Text]
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B. Trigatti, H. Rayburn, M. Vinals, A. Braun, H. Miettinen, M. Penman, M. Hertz, M. Schrenzel, L. Amigo, A. Rigotti, et al. Influence of the high density lipoprotein receptor SR-BI on reproductive and cardiovascular pathophysiology PNAS, August 3, 1999; 96(16): 9322 - 9327. [Abstract] [Full Text] [PDF]
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X. Collet, A. R. Tall, H. Serajuddin, K. Guendouzi, L. Royer, H. Oliveira, R. Barbaras, X.-c. Jiang, and O. L. Francone Remodeling of HDL by CETP in vivo and by CETP and hepatic lipase in vitro results in enhanced uptake of HDL CE by cells expressing scavenger receptor B-I J. Lipid Res., July 1, 1999; 40(7): 1185 - 1193. [Abstract] [Full Text]
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T. M. Forte, M. N. Oda, L. Knoff, B. Frei, J. Suh, J. A. K. Harmony, W. D. Stuart, E. M. Rubin, and D. S. Ng Targeted disruption of the murine lecithin:cholesterol acyltransferase gene is associated with reductions in plasma paraoxonase and platelet-activating factor acetylhydrolase activities but not in apolipoprotein J concentration J. Lipid Res., July 1, 1999; 40(7): 1276 - 1283. [Abstract] [Full Text]
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K. Fluiter, W. Sattler, M. C. De Beer, P. M. Connell, D. R. van der Westhuyzen, and T. J. C. van Berkel Scavenger Receptor BI Mediates the Selective Uptake of Oxidized Cholesterol Esters by Rat Liver J. Biol. Chem., March 26, 1999; 274(13): 8893 - 8899. [Abstract] [Full Text] [PDF]
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Y. Ueda, L. Royer, E. Gong, J. Zhang, P. N. Cooper, O. Francone, and E. M. Rubin Lower Plasma Levels and Accelerated Clearance of High Density Lipoprotein (HDL) and Non-HDL Cholesterol in Scavenger Receptor Class B Type I Transgenic Mice J. Biol. Chem., March 12, 1999; 274(11): 7165 - 7171. [Abstract] [Full Text] [PDF]
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N. Wang, T. Arai, Y. Ji, F. Rinninger, and A. R. Tall Liver-specific Overexpression of Scavenger Receptor BI Decreases Levels of Very Low Density Lipoprotein ApoB, Low Density Lipoprotein ApoB, and High Density Lipoprotein in Transgenic Mice J. Biol. Chem., December 4, 1998; 273(49): 32920 - 32926. [Abstract] [Full Text] [PDF]
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D. V.-v. Bruggen, I. Kalkman, T. van Gent, A. van Tol, and H. Jansen Induction of Adrenal Scavenger Receptor BI and Increased High Density Lipoprotein-Cholesteryl Ether Uptake by in Vivo Inhibition of Hepatic Lipase J. Biol. Chem., November 27, 1998; 273(48): 32038 - 32041. [Abstract] [Full Text] [PDF]
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S. Azhar, A. Nomoto, S. Leers-Sucheta, and E. Reaven Simultaneous induction of an HDL receptor protein (SR-BI) and the selective uptake of HDL-cholesteryl esters in a physiologically relevant steroidogenic cell model J. Lipid Res., August 1, 1998; 39(8): 1616 - 1628. [Abstract] [Full Text]
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N. R. Webb, P. M. Connell, G. A. Graf, E. J. Smart, W. J. S. de Villiers, F. C. de Beer, and D. R. van der Westhuyzen SR-BII, an Isoform of the Scavenger Receptor BI Containing an Alternate Cytoplasmic Tail, Mediates Lipid Transfer between High Density Lipoprotein and Cells J. Biol. Chem., June 12, 1998; 273(24): 15241 - 15248. [Abstract] [Full Text] [PDF]
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E. Reaven, A. Nomoto, S. Leers-Sucheta, R. Temel, D. L. Williams, and S. Azhar Expression and Microvillar Localization of Scavenger Receptor, Class B, Type I (a High Density Lipoprotein Receptor) in Luteinized and Hormone-Desensitized Rat Ovarian Models Endocrinology, June 1, 1998; 139(6): 2847 - 2856. [Abstract] [Full Text] [PDF]
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M. Krieger The "best" of cholesterols, the "worst" of cholesterols: A tale of two receptors PNAS, April 14, 1998; 95(8): 4077 - 4080. [Full Text] [PDF]
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K. Fluiter, D. R. van der Westhuijzen, and T. J. C. van Berkel In Vivo Regulation of Scavenger Receptor BI and the Selective Uptake of High Density Lipoprotein Cholesteryl Esters in Rat Liver Parenchymal and Kupffer Cells J. Biol. Chem., April 3, 1998; 273(14): 8434 - 8438. [Abstract] [Full Text] [PDF]
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A. Rigotti, B. L. Trigatti, M. Penman, H. Rayburn, J. Herz, and M. Krieger A targeted mutation in the murine gene encoding the high density lipoprotein (HDL) receptor scavenger receptor class B type I reveals its key role in HDL metabolism PNAS, November 11, 1997; 94(23): 12610 - 12615. [Abstract] [Full Text] [PDF]
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G. Lambert, N. Sakai, B. L. Vaisman, E. B. Neufeld, B. Marteyn, C.-C. Chan, B. Paigen, E. Lupia, A. Thomas, L. J. Striker, et al. Analysis of Glomerulosclerosis and Atherosclerosis in Lecithin Cholesterol Acyltransferase-deficient Mice J. Biol. Chem., April 27, 2001; 276(18): 15090 - 15098. [Abstract] [Full Text] [PDF]
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