Lipid Metabolism

Lipid metabolism is a dynamic biological process that includes lipid synthesis,

accumulation, distribution to various specific tissues, and excretion [1].

From: Advances in Clinical Chemistry, 2018

Related terms:

Lipid, Protein, Insulin, Liver, Gene, Cholesterol, Obesity

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Learn more about Lipid Metabolism

Conceptual Background and Bioen-

ergetic/Mitochondrial Aspects of On-
cometabolism
Juliet Goldsmith, ... Jayanta Debnath, in Methods in Enzymology, 2014

5.1.4 Lipids
Lipid metabolism is altered in cancer—tumor cells reactivate de novo lipid synthesis,
ATP-citrate lyase is required for transformation in vitro, cholesterol synthesis in
prostate cancer is increased, and fatty acid oxidation is an important source of energy
for prostate cancer cells (Santos & Schulze, 2012). Autophagy in the specific form
of lipophagy is important for the degradation of lipid droplets in the adipose tissue
(Singh & Cuervo, 2012), and autophagy regulates lipid metabolism in hepatocytes as
triglyceride hydrolysis is impaired in Atg5−/− cells (Singh et al., 2009). Whether these
processes affect tumor lipid metabolism requires further study.

Additionally, autophagy impacts lipid metabolism by altering the mitochondrial

number. Atg7 deleted, p53 mutant cells in a KRAS-driven NSCLC model have in-
tracellular lipid accumulation because of increased dysfunctional mitochondria that
compromises fatty acid oxidation, suggesting that autophagy is crucial to maintain
lipid metabolism in KRAS and p53 mutant cells. This prevents the efficient growth
of tumor cells and turns them into lipid cysts instead of tumors (Guo et al., 2013).

> Read full chapter

Advances in Clinical Chemistry

Yongzhi Zeng, ... Guanghui Yi, in Advances in Clinical Chemistry, 2018

1 Introduction
Lipid metabolism is a dynamic biological process that includes lipid synthesis,
accumulation, distribution to various specific tissues, and excretion [1]. The mol-
ecular machinery of lipid homeostasis is triggered by a series of well-character-
ized transporters and enzymes that are differentially regulated depending on the
requirements of the organism [2]. In patients with hyperlipidemia, many signaling
pathways implicated in lipid metabolism are dysregulated, leading to alteration of
circulating lipid levels. The discovery that key components of these pathways are
targeted by lncRNAs has attracted our attention to the role of lncRNAs as lipid
metabolism regulators, with therapeutic applications for lipid metabolism-related
diseases, including atherosclerosis, obesity, Alzheimer's disease, type II diabetes, and
other metabolic diseases [3].

Long noncoding RNAs are emerging as a unique group of RNA transcripts more
than 200 nucleotides in length with no protein-coding potential [4]. lncRNAs func-
tion as signals, decoys, guides, and scaffolds to modulate the expression of target
genes at both the transcriptional and posttranscriptional level [5–7]. While the
biological functions of most lncRNAs are poorly characterized, numerous gain- and
loss-of-function studies using in vitro and in vivo animal models have extended our
knowledge of the role of lncRNAs in the initiation and progression of disease. Recent
studies have found that lncRNAs participate in metabolic regulation, inflammation,
immunity and vascular function. Here, we outline recent investigations that provide
novel insights into the mechanisms by which lncRNAs act to influence lipid and
lipoprotein metabolism, with possible implications for lipid metabolism-related
disease processes.

> Read full chapter

Biochemistry and Metabolism of Toxo-

plasma gondii
Isabelle Coppens, ... Stanislas Tomavo, in Toxoplasma Gondii (Second Edition), 2014

Lipid Metabolism 269

8.5.1Fatty Acids 2708.5.1.1Fatty Acid Biosynthetic Pathways – Generalities 270-

8.5.1.2Fatty Acid Synthesis in Toxoplasma 2708.5.1.3Fatty Acid Salvage by
Toxoplasma 273
8.5.2Glycerophospholipids 2748.5.2.1Phospholipid Biosynthetic Pathways – Gen-
eralities 2748.5.2.2Phospholipid Composition and Physiological Relevance in
Toxoplasma 2748.5.2.3Phospholipid Synthesis in Toxoplasma 2768.5.2.4Phos-
pholipid Salvage by Toxoplasma 276
8.5.3Glycerolipids 2778.5.3.1Glycerolipid Biosynthetic Pathways – Generalities -
2778.5.3.2Glycerolipid Synthesis in Toxoplasma 278
8.5.4Sterols and Steryl Esters 2788.5.4.1Sterol Lipid Biosynthetic Pathways – Gen-
eralities 2788.5.4.2Sterol Salvage and Transport in Toxoplasma 2798.5.4.3-
Sterol Storage in Toxoplasma 279
8.5.5Sphingolipids 2808.5.5.1Sphingolipid Biosynthetic Pathways – Generali-
ties 2808.5.5.2Sphingolipid Synthesis in Toxoplasma 2808.5.5.3Sphingolipid
Salvage by Toxoplasma 281
8.5.6Isoprenoid Derivatives 2828.5.6.1Isoprenoid Biosynthetic Pathways – Gen-
eralities 2828.5.6.2Isoprenoid Synthesis in Toxoplasma 2828.5.6.3Isoprenoid
Salvage by Toxoplasma 282

> Read full chapter

Lipid Disorders in Obesity

Manoj Wickramasinghe MBBS, Jolanta U. Weaver MRCS, FRCP, PhD, CTLHE, in
Practical Guide to Obesity Medicine, 2018

Normal Lipid Metabolism

Lipid metabolism involves a number of key enzymes and subtypes of lipid fractions
and lipoproteins. These include lipoprotein lipase (LPL), TAGs, cholesterol (CH),
HDL, LDL, very-low-density lipoprotein (VLDL), and chylomicrons. TAGs are the
main constituent of dietary lipids and their principal function is to act as energy
reservoir stored in adipocytes. Lipids are insoluble in blood and are packaged in
lipoproteins, which act to transport lipids in the blood. Lipoproteins are classified by
their density and size.

To understand the pathogenesis of adiposity and lipid metabolism, it is critical to

understand normal lipid metabolism. To do this it is helpful to look at three of the key
pathways: the dietary component, endogenous component, and finally the reverse
cholesterol transport pathway.

Exogenous Pathway
Following digestion and absorption of dietary fat, TAG and CH combine with protein
and phospholipids to form chylomicrons in the epithelial layer of the small intes-
tine.5 These chylomicrons are then absorbed into the lymphatic system where they
enter the circulation via the thoracic duct. Here they are transported to the rest of the
body to be stored in adipose tissue. Endothelial LPL cleaves TAGs from chlylomicrons
to form FFAs and glycerol, which can enter skeletal muscle and adipose tissue,
leaving behind chylomicron remnants that are recycled by the liver (see Fig. 10.1).

Fig. 10.1. Lipid Metabolism.Summary of the three key pathways of lipid metabo-
lism: exogenous pathway, endogenous pathway, and reverse cholesterol transport.
(A) The exogenous pathway: action of lipoprotein lipase (LPL) on chylomicrons to
form free fatty acids (FFAs). (B) The reverse cholesterol transport pathway: action
of high-density lipoprotein (HDL) in collecting peripheral cholesterol. (C) The en-
dogenous pathway: role of LPL in cleaving very-low-density lipoproteins (VLDL) to
form intermediate density lipoproteins (IDLs), taken up by the liver or low-density
lipoproteins (LDLs) and FFAs.

Endogenous Pathway
VLDL is synthesized by the liver and released into the systemic circulation. In the
tissues LPL cleaves TAGs from the VLDL to release fatty acids, which are taken up
by myocytes for energy or by adipose tissue for storage. As LPL cleaves TAGs, the
cholesterol concentration within the lipoprotein increases and becomes a smaller,
denser lipoprotein named “intermediate density lipoproteins” (IDL)—the action of
LPL may continue to form LDL, a smaller denser particle than IDL.

Reverse Cholesterol Transport

It involves the movement of cholesterol from non-hepatic peripheral tissues back to
the liver. This is mainly regulated by the ATP-binding cassette transporter on HDL,
which allows the transfer of cholesterol onto the HDL.

> Read full chapter

Methods of Adipose Tissue Biology,

Part B
Lee D. Roberts, ... Julian L. Griffin, in Methods in Enzymology, 2014

Abstract
Lipid metabolism is central to the function of white adipose tissue, with the tissue
having a central role in storing triacylglycerides following feeding and releasing
free fatty acids and monoacylglycerides during periods of fasting. In addition, lipid
species have been suggested to play a role in lipotoxicity and as signaling molecules
during adipose tissue inflammation. This chapter details how mass spectrometry
(MS) can be used to profile a range of lipid species found in adipose tissue. The
initial step required in any MS-based approach is to extract the lipid fraction from
the tissue. We detail one commonly used method based on the Folch extraction
procedure. The total fatty acid composition of the lipid fraction can readily be
defined using gas chromatography–MS, and we provide a method routinely used
for rodent and human adipose tissue samples. However, such approaches do not
provide insight into what lipid classes the various fatty acids are associated with. To
better understand the global lipid profile of the tissue, we provide a general-purpose
liquid chromatography–MS-based approach useful for processing phospholipids,
free fatty acids, and triacylglycerides. In addition, we provide a method for profiling
eicosanoids, a class of important lipid-signaling molecules, which have been im-
plicated in white adipose tissue inflammation in rodent models of obesity, insulin
resistance, and type 2 diabetes.
> Read full chapter

Diabetes Mellitus in Animals

Deborah S. Greco, in Nutritional and Therapeutic Interventions for Diabetes and
Metabolic Syndrome, 2012

Ketoacidotic, Hyperosmolar or Complicated Diabetes Mellitus:

Pathophysiology and Clinical Signs
Lipid metabolism in the liver becomes deranged with insulin deficiency and non-es-
terified fatty acids are converted to acetyl-CoA rather than being incorporated
into triglycerides. Acetyl-CoA accumulates in the liver and is converted into ace-
toacetyl-CoA and then ultimately to acetoacetic acid. Finally, the liver starts to gen-
erate large amounts of ketones including acetoacetic acid, beta-hydroxybutyrate and
acetone. As insulin deficiency culminates in DKA, accumulation of ketones and lactic
acid in the blood and loss of electrolytes and water in the urine results in profound
dehydration, hypovolemia, metabolic acidosis, and shock. Ketonuria and osmotic
diuresis caused by glycosuria result in urinary sodium and potassium loss which
exacerbates hypovolemia and dehydration. Nausea, anorexia and vomiting, caused
by stimulation of the chemoreceptor trigger zone via ketonemia and hyperglycemia,
contribute to the dehydration caused by osmotic diuresis. Dehydration and shock
lead to prerenal azotemia and a decline in glomerular filtration rate (GFR). Declining
GFR leads to further accumulation of glucose and ketones in the blood. Stress
hormones such as cortisol and epinephrine contribute to the hyperglycemia in a
vicious cycle. Eventually severe dehydration may result in hyperviscosity, throm-
boembolism, severe metabolic acidosis, renal failure, and finally death.

The most common historical findings in cats and dogs with diabetic ketoacidosis are
anorexia (61%), weakness, depression, and vomiting.2 Physical examination findings
may include shock, depression, tachypnea, dehydration, weakness, vomiting, and
occasionally, a strong acetone odor on the breath. Cats can present recumbent or
comatose; this may be a manifestation of mixed ketotic hyperosmolar syndrome. In
cats, 33% exhibited clinical icterus at presentation.2

> Read full chapter

Diabetes Mellitus in Animals

Deborah S. Greco, in Nutritional and Therapeutic Interventions for Diabetes and
Metabolic Syndrome (Second Edition), 2018

Ketoacidotic, Hyperosmolar, or Complicated Diabetes Mellitus:

Pathophysiology and Clinical Signs
Lipid metabolism in the liver becomes deranged with insulin deficiency and non-
esterified fatty acids are converted to acetyl-CoA rather than being incorporated
into triglycerides. Acetyl-CoA accumulates in the liver and is converted into ace-
toacetyl-CoA and then ultimately to acetoacetic acid. Finally, the liver starts to
generate large amounts of ketones, including acetoacetic acid, beta-hydroxybu-
tyrate, and acetone. As insulin deficiency culminates in diabetic ketoacidosis (DKA),
accumulation of ketones and lactic acid in the blood and loss of electrolytes and
water in the urine result in profound dehydration, hypovolemia, metabolic acidosis,
and shock. Ketonuria and osmotic diuresis caused by glycosuria result in urinary
sodium and potassium loss, which exacerbates hypovolemia and dehydration. Nau-
sea, anorexia, and vomiting, caused by stimulation of the chemoreceptor trigger
zone via ketonemia and hyperglycemia, contribute to the dehydration caused by
osmotic diuresis. Dehydration and shock lead to prerenal azotemia and a decline
in glomerular filtration rate (GFR). Declining GFR leads to further accumulation of
glucose and ketones in the blood. Stress hormones such as cortisol and epinephrine
contribute to the hyperglycemia in a vicious cycle. Eventually, severe dehydration
may result in hyperviscosity, thromboembolism, severe metabolic acidosis, renal
failure, and finally death.

The most common historical findings in cats and dogs with DKA are anorexia (61%),
weakness, depression, and vomiting.2 Physical examination findings may include
shock, depression, tachypnea, dehydration, weakness, vomiting, and occasionally
a strong acetone odor on the breath. Cats can present recumbent or comatose;
this may be a manifestation of mixed ketotic hyperosmolar syndrome. In cats, 33%
exhibited clinical icterus at presentation.2

> Read full chapter

Disorders of Lipid Metabolism

Clay F. Semenkovich, ... Ira J. Goldberg, in Williams Textbook of Endocrinology
(Thirteenth Edition), 2016

Integrative Physiology of Lipid Metabolism

Lipid metabolism is characterized by a dynamic flux of multiple lipid species from the
external environment to the liver, from the liver to peripheral tissues, from peripheral
tissues back to the liver, and eventually back to the external environment through the
excretion of bile acids. Integrated views of the major pathways involved are shown
in Figures 37-14 and 37-15.

Figure 37-14. General scheme summarizing the major pathways involved in the me-
tabolism of chylomicrons synthesized by the intestine and very low density lipopro-
tein (VLDL) synthesized by the liver. ApoB, apolipoprotein B; ApoE, apolipoprotein
E; FFA, free fatty acid; HL, hepatic lipase; IDL, intermediate-density lipoprotein; LPL,
lipoprotein lipase.

(Modified from Mahley RW. Biochemistry and physiology of lipid and lipoprotein
metabolism. In: Becker KL, ed. Principles and Practice of Endocrinology and Metabo-
lism, 2nd ed. Philadelphia: JB Lippincott; 1995:1369-1378.)
Figure 37-15. Role of high-density lipoprotein (HDL) in the redistribution of lipids
from cells with excess cholesterol to cells requiring cholesterol or to the liver for ex-
cretion. The reverse cholesterol transport pathway is indicated by arrows (net transfer
of cholesterol from cells to HDL, then to LDL and liver). ApoE, apolipoprotein E; CE,
cholesteryl ester; CETP, cholesteryl ester transfer protein; IDL, intermediate-density
lipoprotein; LCAT, lecithin:cholesterol acyltransferase; LDLR, low-density lipoprotein
receptor; SR-BI, scavenger receptor class B, type 1; Tg, triglyceride; VLDL, very low
density lipoprotein.

Exogenous Lipid Transport

Dietary fat and cholesterol (see Fig. 37-14, top left) absorbed by the duodenum and
proximal jejunum are used to generate chylomicrons that are secreted at the lateral
borders of enterocytes and enter mesenteric lymphatics. They access the plasma via
the thoracic duct and are rapidly metabolized by LPL to yield chylomicron remnants.
These are taken up by remnant receptors (LRP1/HSPG) and by LDL receptors in the
liver. Free fatty acids liberated by the action of LPL are available to adipose tissue for
storage and to other tissues (e.g., skeletal muscle, heart) for use as energy substrates.

Endogenous Lipid Transport

Lipid derived from remnants and from lipolysis of adipose tissue is reassembled in
the liver (see Fig. 37-14, bottom left) as VLDL particles, which are secreted into the
plasma. Abnormal lipid metabolism in insulin resistance is mediated in large part
by overproduction of VLDL, an event that occurs through disruption of signaling
downstream of the insulin receptor and the insulin receptor substrate (IRS) adapter
proteins. VLDL particles are metabolized by LPL to yield IDL particles, which are
metabolized by LPL and hepatic lipase to yield LDL particles. Thus, LDL is derived
from VLDL, which helps explain why treatment to lower triglycerides (carried by
VLDL) is frequently associated with at least transient increases in LDL. IDL can be
taken up by the liver through an apoE-dependent process, and LDL is taken up by
the liver through the binding of apoB100 to LDL receptors. Small VLDL particles,
IDL particles, and LDL particles may be taken up by peripheral tissues to deliver
nutrients, cholesterol, and fat-soluble vitamins. When present in excess, each of
these lipoproteins may be atherogenic.

Reverse Cholesterol Transport and Dysfunctional HDL

Cholesterol cannot be metabolized by peripheral tissues and must be returned
to the liver for excretion. This process, known as reverse cholesterol transport, is
dependent on HDL and its precursors and is depicted in Figure 37-15. Excess
cholesterol in tissues can be effluxed either to lipid-poor apoAI, mediated by the
protein transporter ABCA1 (adenosine triphosphate–binding cassette transporter
1), or to nascent HDL particles, mediated by ABCG1. Efflux from cultured cells to
human plasma as a biomarker of cardiovascular risk39 is thought to mostly reflect
the activity of ABCA1. There is also evidence that cholesterol can be acquired by
HDL without the assistance of transporters by following a concentration gradient
at the cell surface. LCAT esterifies HDL-associated cholesterol to form cholesteryl
ester and induce the maturation of HDL. HDL particles have three pathways for
transporting sterols to the liver. First, they can directly bind to SR-BI (CLA-1) at the
liver, which induces cholesteryl ester delivery through a mechanism involving lateral
lipid transfer and not receptor internalization. Second, cholesteryl esters can be
transferred to apoB-containing lipoproteins by CETP, and these particles can deliver
cholesterol to the liver through the LDL receptor. Third, a small portion of HDL can
acquire apoE and bind to the liver LDL receptor. Once in the liver, cholesterol is
converted to bile acids for excretion.

HDL2 particles are partially depleted of cholesteryl esters and enriched in triglyc-
erides through the activity of CETP, which renders them suitable as substrates for
hepatic lipase. Hepatic lipase hydrolyzes the triglyceride-enriched HDL2 particles
and regenerates HDL3, yielding particles that are again suited to accept cholesterol
from peripheral cells.

Because cholesterol is the principal component of atherosclerotic plaque, it is rea-

sonable to pursue the notion that atherosclerosis could be treated by promoting
the efflux of cholesterol from lesions. HDL participates in this process, but stat-
ic levels of HDL cholesterol are poor predictors of reverse cholesterol transport.
Measurements of the rate of flux of cholesterol from the periphery to the liver,
which may be possible in humans, would represent a better predictor of beneficial
therapies. In vitro HDL efflux capacity represents an initial step toward assessing
reverse cholesterol transport.39 The presence of high levels of HDL particles that
have been modified to prevent their capacity to promote cholesterol efflux would
not be expected to decrease vascular risk. Such dysfunctional particles may explain
why some HDL-elevating interventions have not been associated with decreased
cardiovascular disease.

In addition to participating in reverse cholesterol transport, HDL has other prop-

erties that could be impaired by a variety of processes leading to a dysfunctional
particle. They include the induction of endothelial nitric oxide synthase, the transport
of proteins involved in the acute phase response and inflammation, and the suppres-
sion of thrombosis through induction of prostacyclin (which decreases thrombin
production via the protein C pathway and decreases platelet activation).

> Read full chapter

Disorders of Lipid Metabolism

Clay F. Semenkovich, in Goldman's Cecil Medicine (Twenty Fourth Edition), 2012

Reverse Cholesterol Transport and High-Density Lipoprotein

Metabolism
Lipid metabolism is dynamic. At the same time lipoproteins are being processed to
modify their nonpolar lipids, particles are interacting with one another, exchang-
ing surface materials, apolipoproteins, and nonpolar lipids. HDL is an important
reservoir for components cast off during the metabolism of other lipoproteins as
well as lipids discarded by cells. Nascent HDL is generated by the liver and intestine
as a phospholipid disc containing apo AI and apo AII. It accepts unesterified (free)
cholesterol (UC in Fig. 213-2) and phospholipids shed from cells. This unesterified
cholesterol is converted to cholesteryl ester by the action of LCAT and stored in
the center of the disc, allowing it to become a spherical particle. The particle is
further modified as a consequence of the action of LPL on triglycerides in apo
B–containing lipoproteins. As the core triglycerides of VLDL are metabolized, the
particle collapses, leaving redundant surface lipids (phospholipid in the form of
lecithin and unesterified cholesterol) and excess apolipoproteins such as apo CII, apo
CIII, and apo E, which are transferred to HDL. LCAT again esterifies the cholesterol
to increase the content of cholesteryl ester in the HDL.
FIGURE 213-2. Reverse cholesterol transport and high-density lipoprotein (HDL)
metabolism.Unesterified cholesterol (UC) in peripheral cells can be transferred to
HDL and esterified by lecithin-cholesterol acyltransferase (LCAT). This cholesterol
ester (CE) in HDL can be transferred to the liver directly through scavenger receptor
B1 (SR-B1). Alternatively, it can be transferred to apolipoprotein B100–containing
lipoproteins in exchange for triglycerides (TG) through the action of cholesteryl ester
transfer protein (CETP). IDL = intermediate-density lipoprotein; LDL = low-densi-
ty lipoprotein; LDLR = low-density lipoprotein receptor; VLDL = very low density
lipoprotein.

Reverse cholesterol transport is the beneficial process by which cholesterol present in

peripheral cells, such as foam cells in a growing atherosclerotic lesion, is transported
back to the liver for excretion. There are at least two well-defined pathways mediating
this transfer. First, after accepting cholesterol from peripheral cells and esterifying
it through the action of LCAT, HDL can interact directly with the liver by binding to
SR-B1 and transferring cholesteryl ester to the hepatocyte. Second, HDL can transfer
cholesteryl ester to apo B100–containing lipoproteins such as VLDL (see bottom
of Fig. 213-2) through the action of CETP. This cholesteryl ester can ultimately be
transported to the liver after conversion of VLDL to IDL to LDL and uptake by the
LDL receptor (LDLR in Fig. 213-2). This pathway is not direct because the transfer
of cholesteryl ester to apo B–containing lipoproteins results in cholesterol-enriched
particles that may be taken up by foam cells in atherosclerotic plaques before being
cleared by the liver. Humans with genetic defects in CETP have high HDL levels and
appear to be healthy. An inhibitor of CETP, torcetrapib, increases HDL cholesterol
but also increases adverse events, perhaps in part related to its adverse effects
on blood pressure. The benefits of raising HDL with medications are uncertain; a
meta-analysis of more than 100 randomized trials showed no reduction in coronary
heart disease events by simply increasing HDL.1
> Read full chapter

The Zebrafish: Cellular and Develop-

mental Biology
Shiu-Ying Ho, ... Steven A. Farber, in Methods in Cell Biology, 2004

III Pharmacological Studies Using Zebrafish

Lipid metabolism in zebrafish larvae can be successfully modulated by a number of
pharmacological means. We have previously shown that atorvastatin (Lipitor) can
block the accumulation of fluorescent lipids in the gall bladder of larval zebrafish
(Farber et al., 2001). Lipitor is a widely prescribed medication for the treatment of
hypercholesterolemia, and is one of the most economically successful drugs ever
developed. Lipitor belongs to the statin family of 3-hydroxyl-3-methylglutaryl-CoA
reductase (HMGCoAR) inhibitors (Fig. 6). It has been reported that in Drosophila,
reduced HMGCoAR activity results in primordial germ cell (PGCs) migration defects
(Van Doren et al., 1998). We therefore examined both zebrafish overall morphological
development and PGC migration following statin-mediated HMGCoAR inhibition.
PGCs migration in zebrafish embryos can be visualized in late somite-stage embryos
by injecting in vitro transcribed capped gfp-nanos mRNA at the one-cell stage.
Both the gfp-nanos message and protein are stabilized only in PGCs such that
they maintain their fluorescence throughout early development, facilitating detailed
study of PGC migratory behavior and ultimately gonad formation (Doitsidou et
al., 2002). When zebrafish embryos are soaked in mevinolin (Lovastatin) and sim-
vastatin (Zocor), embryos exhibited developmental arrest, blunted axis elongation,
misshapen somites, and head and axial necrosis. Further, PGC migration was pro-
foundly perturbed with a similar dose-response profile (Fig. 7). Embryos treated with
the more hydrophobic statin, Lipitor, exhibited PGCs migration defects and only
mild morphologic abnormalities (Fig. 8).
Fig. 6. Cholesterol and isoprenoid biosynthesis pathway (Thorpe et al., 2004).

Fig. 7. Embryos treated with statins exhibit developmental defects (Thorpe et al.,
2004). (A) Wild-type embryos (24 hpf ). (B) Embryos soaked in a low dose of mevinolin
(0.06 μM) have a slight tail kink. (C) Exposure to higher doses of mevinolin (1.2 μM)
results in blunted axis elongation, necrosis, and developmental arrest. (D) Simvas-
tatin-treated embryos (2.0 μM) exhibit similar defects as in (C). (E) Dose-response
of mevinolin on developmental arrest (mean ± SEM, n = 3). (F) Dose-response of
simvastatin on developmental arrest (mean ± SEM, n = 3).

Fig. 8. Migration of primordial germ cells (PGCs) is altered by atorvastatin treatment

(Thorpe et al., 2004). (A) A single fluorescent image from a time lapse (1-somite stage,
first capture) of the movement of PGCs in a wild-type embryo injected at the 1 cell
stage with gfp-nanos mRNA (1–2 nl, 60 ng ul). (B) Embryos (1-somite stage) soaked
in atorvastatin (10 mM) and injected with gfp-nanos mRNA exhibit ectopic PGCs
that fail to cluster. (C) Later stage embryos (22-somite stage) exhibit many ectopic
germ cells after atorvastatin (10 mM) treatment. (D) Brightfield image of an embryo
(24 hpf ) injected with gfp-nanos mRNA and treated with atorvastatin (10 mM) that
exhibits only a mild tail defect. (E) Analysis of the effect of atorvastatin treatment
on PGC migration at different developmental stages. The number of embryos at a
given stage with more than 2 ectopic PGCs were determined and expressed as a
percent of the total number of embryos examined (n = 64 – 84 for each stage). (F)
The effect of atorvastatin on numbers of ectopic PGCs is dose dependent. Embryos
were soaked in atorvastatin (10 μM). At 24 hpf, embryos were each scored on the
degree of ectopic PGCs (level 1 embryo had a wild-type single gonadal cluster and
a level 4 embryo had no discernible cluster). Data represent the mean ± SEM from
3 to 4 experiments dose.

To determine which downstream products of HMGCoAR mediate these defects, we

pioneered a “block in rescue” approach by injecting putative biochemical pathway
intermediates downstream of the pharmacologically inhibited enzyme. We initially
established the validity of this approach by demonstrating that an injection of meval-
onate, the product of HMGCoAR, completely rescued all the phenotypes associated
with statin treatment (Fig. 9). Similar experiments using intermediates downstream
of mevalonate (see Fig. 6) indicated that the prenylation pathway was mediating the
effect of statins on PGCs. We tested this hypothesis by treating gfp-nanos mRNA
injected embryos with selective geranylgeranylation or farnesylation inhibitors. High
doses of the selective farnesyl transferase inhibitors L-744 or FTI-2153 (Crespo et
al., 2001; Sun et al., 1999) had no effect on PGC migration. Embryos treated with
geranylgeranyl transferase I (GGTI) inhibitor (GGTI-2166) exhibited a strong PGC
migration phenotype with only mild morphological defects manifested as a slight
disruption of the notochord (Fig. 10). These data suggest that protein prenylation,
specifically by GGTI, is required for correct PGC migration (Thorpe et al., 2004).
Fig. 9. Injection of isoprenoid intermediates abrogates the effects of statins (Thorpe
et al., 2004). (A) Uninjected embryos show severe developmental defects after 24 h
of mevinolin (1.2 μM) treatment. (B) Embryos injected at early cell stages (1 to 16
cell stage) with mevalonate (1 to 2 nl, 0.5 M) and then soaked in mevinolin (1.2
μM, 24 h) exhibit normal morphology. (C) Mevalonate injection rescues the somatic
defects observed in embryos treated with statins. Embryos at early cell stages were
injected with mevalonate as in (B) and soaked overnight in statin drugs: mevinolin
(1.2 μM), simvastatin (2.0 μM), and atorvastatin (10 μM). At 24 hpf, developmental
defects were scored. Data represent the MEAN ± SEM from 3 to 4 experiments. (D)
The appearance of ectopic PGCs after statin treatment is prevented by mevalonate
injections. Embryos at early cell stages were injected with gfp-nanos mRNA and
mevalonate as in (B) and soaked overnight in statin drugs [mevinolin (1.2 μM),
simvastatin (2.0 μM), and atorvastatin (10 μM)]. At 24 hpf, the PGC score was
determined. Data represent the Mean ± SEM from 3 to 4 experiments. (E) Embryo's
injected at the 1- to 16-cell stage with farnesol (1 to 2 nl, 1 M), geranyl geraniol (0.5 to
1.0 nl, 1 M) or mevalonate (1 to 2 nl, 0.5 M) then soaked overnight with atorvastatin
(10 μM) show normal PGC migration. Statistics were performed using ANOVA with
a post hoc test that utilizes a Bonferroni correction. Data represents MEAN ± SEM,
p < 0.01 difference from atorvastatin alone.

Fig. 10. GGT1 activity is required for PGC migration (Thorpe et al., 2004). (A) A
fluorescent image of a wild-type embryo (24 hpf ) injected with gfp-nanos mRNA at
the 1-cell stage reveals a cluster of PGCs. (B) A brightfield image of the embryo in
(A). (C) A fluorescent image of an embryo (24 hpf ) that was injected with gfp-nanos
mRNA at the 1-cell stage and immediately soaked in the geranylgeranyl transferase
inhibitor (GGTI-2166; 40 μM), resulting in ectopic migration of many PGCs. (D) A
brightfield image of the embryo in (C) showing a mild notochord defect (arrowhead).
(E) Dose response of GGTI-2166 on PGC migration defect. Data represent the MEAN
± SEM from 3 to 6 experiments for each dose.

> Read full chapter

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