THE JOURNAL OF B~LOGICAL CHEMISTRY

Vol. 248, No. 1. Issue of January 10,~~. 270-278,1973

Printed

in

U.S.A.

The Effects of Starvation

Lipid Metabolism
in Viva
Perfused
Rat Liver

and Refeeding
and in the

THE RELATIOSSHIP
BETWEEN
REGULATION
OF KETOGENESIS

ACID

FATTY

OXIDATION

on Carbohydrate

AND

ESTERIFICATION

IN THE

(Received for publication,

J.

DENIS

m!GARRY,

J~RGEN

M.

n/IEIER,

From. ihe Departments

of Internal
Medicine
School at Dallas, Dallas, Texas 75,935

AND

D~IEL

and Biochemistry,

The time course of changes in a variety of physiological

parameters
concerned with carbohydrate
and lipid metabolism has been studied both in vivo and in the isolated perfused
liver during induction
and reversal of starvation
ketosis in
the rat.
The data obtained
demonstrate
that surprisingly
brief periods of starvation
and refeeding
exert dramatic
effects on glucose and fatty acid metabolism
in the intact
animal and that generally synchronous
changes occur in the
ketogenic and gluconeogenic
capacities of the perfused liver.
In agreement with previous findings it was shown that the
enhanced conversion of labeled oleate into ketone bodies by
livers from fasted rats was associated with a concomitant
depression
in its incorporation
into triglycerides,
and that
the antiketogenic
effect of lactate was accompanied
by a
diversion of the fatty acid from the /3 oxidation sequence into
the esterification
pathway.
The key observation,
however,
was that blockade
of long chain fatty acid oxidation
by
(+)-decanoylcarnitine,
an inhibitor
of long chain acylcarnitinetransferase,
stopped ketone body formation
and
acutely changed the pattern of metabolism
of oleic acid in
livers from fasted rats to that exhibited by livers from normal
animals, i.e. the fatty acid was now virtually
completely
esterified.
The data are consistent with the view that hepatic fatty
acid oxidation and ketogenesis
are under strict dietary and
hormonal control exerted primarily
by regulation
of an early
step in the oxidative sequence, probably the acylcarnitinetransferase reaction.
The possibility
is also raised that the
effects of lactate and other antiketogenic
agents are related
to interactions
at this site.

The strikiiig
enhanccmcnt
in hepatic gluconcogenesis
and
ketogenesis characteristic of the fasting state is well documented.
by United
Awardee

States Public
5-K3-AM

9968,

270

June 26,1972)

FOSTERI

The

University

of Texas Southwestern

Medical

In recent years many of the investigations into the regulation of

these processes have utilized the isolated perfused rat liver
technique and have been based upon comparative studies of the
metabolism of livers taken from well fed or fasted animals (e.g.
l-5).
In some of these studies a starvation period of 24 hours
was routinely used (2-4) whereas in others the animals were
fasted for 48 hours (1, 5). In general, however, it appears that
both time periods result in maximal increases above control
levels in the gluconeogenic and ketogenic capacities of the liver.
The question is raised, therefore, as to what length of starvation
is required to bring about the full activation of these two metabolic events and what type of kinetics is displayed during the
induction process. In an attempt to answer these questions we
have examined in some detail the effects of different periods of
starvat,ion and refeeding on a variety of physiological parameters
concerned wibh carbohydrate and lipid metabolism in the intact
rat. 1 crit.ical issue was whether the isolated perfused liver
system adequately reflects metabolic events occurring in viva
during the onset and reversal of starvation ketosis. In a parallel
series of experiments, therefore, we have tested the effects of
starvation and refeeding on the ability of the isolated organ to
synthesize glucose and ketone bodies when perfused with the
appropriate
substrates.
A second objective of the present investigation
was to gain
further insight into the relationship between fatty acid oxidation
and esterification with regard to the over-all regulation of ketone
body synthesis in the liver.
Previous studies (5, 6) have clearly
shown that the enhanced oxidation of long chain fatty acids induced by starvation is quantitatively
balanced by a concomitant
decrease in their incorporation
into triglycerides and other liver
lipids.
These findings were consistent with the widely held view
that the fraction of a long chain fatty acid load undergoing oxidation depends primarily upon the efficiency of the esterification
pathway which in turn has generally been considered to be
governed principally
by the availability
of cr-glycerophosphate
(sn-glycero-3-phosphate).
We have pointed out, however, that
this concept might in fact be incorrect (5, 7, 8). This note of
caution was prompted by the lack of correlation between hepatic a-glycerophosphate
levels and the rates of ketogenesis
observed both in the perfused rat liver (5) and in in viva studies

Downloaded from http://www.jbc.org/ by guest on August 25, 2016

SUMMARY

* This investigation
was slipported
Health Service Grant CA-08269.
$ Research
Career Development
United States Public IIealth Service.

W.

and

271
(9). Moreover, the view that depressed triglyceride
synt,hesis
is the primary mechanism for the shunting of fatty acids into
the oxidative pathway in the ketotic state is difficult to reconcile
with the situation commonly found in experimental
diabetic
ketoacidosis, where increased hepatic concentration
of fat occurs
in the face of higher rates of ketone production than are seen in
starvation (7, IO).
The data outlined below illustrate
that surprisingly
brief
periods of starvation and refeeding exert striking effects on glucose and fatty acid metabolism in the intact rat and that a remarkable degree of synchrony exists between the events taking
place in vivo and those observed in the isolated perfused liver.
In addition, evidence will be presented to support the thesis that
in the fasted state no fundamental defect is present in the triglyceride-synthesizing
capacity of the liver, and that the marked
enhancement of ketogenesis is due to preferential flux of available
fatty acid carbon through the /? oxidation sequence, the latter
event probably resulting from the activation of an early step in
the oxidative pathway.
PROCEDURE

Animals-Male
Sprague-Dawley
rats weighing 100 to 120 g
were used in all experiments.
Animals were fed a diet containing 58.5% sucrose, 21% casein, and less than lye fat (General
Biochemicals,
Chagrin Falls, Ohio).
The term normal
as
used in the text refers to rats that had free access to food up to
the time of commencement of experiments which was generally
between 9:30 and 10:00 a.m. All fasted rats were deprived of
food for the indicated time periods. When animals were fasted
for shorter times (6 to 12 hours) they received 2 ml of a balanced
liquid diet containing
34% glucose (Diet ~~116 EC, General
Biochemicals) by stomach intubation
as a final feeding in order
to standardize the period of fasting.
Animals used in refeeding
studies received 3 ml of the liquid diet initially followed by l-ml
portions at hourly intervals until used for experiments.
Analyses on Blood and Liver from Intact Rats-Livers
and
blood samples were removed and plasma was analyzed for ketone
bodies, free fatty acids, glucose, and insulin as described earlier
(7). Liver glycogen was determined by the method of Good
et al. (11).
Liver Perfusion-Livers
were perfused with recirculating
medium using the apparatus and techniques described in previous
reports (5, 12). As routinely used in these studies aged human
erythrocytes were washed with 0.9% sodium chloride and Krebs
bicarbonate buffer, pH 7.4, before being suspended in the latter
to a hematocrit of 40%. This was mixed with an equal volume
of a solution containing
8% bovine albumin in Krebs bicarbonate buffer, pH 7.4, to provide the final perfusion fluid.
In
the present series of experiments it was observed that the initial
lactate concentration
in the cell-free fluid was regularly in the
region of 3 mM. Because of the known antiketogenic
properties
of lactate, in some experiments it was desirable to remove it
from the medium.
Since the lactate was known to originate
from the erythrocytes, this was accomplished by dialyzing the
red cells for 24 hours against 0.9% sodium chloride at 4 prior
to use.
In all of the experiments to be described, sufficient oleic acid
was included in the initial pool to give a fatty acid concentration
in the cell-free fluid of approximately
0.7 InM.
This concentration was maintained throughout
the experimental period by the
infusion of a solution containing 12.5 mM oleate and 10% albumin dissolved in 0.9% NaCl solution, pH 7.4, at a rate of 75 ~1
per min. In experiments using [1-14CJoleic acid 0.4 ml of a 5%

RESULTS

and Refeeding on Carbohydrate and Lipid

Xetabolism in Vivo-The
first series of experiments was designed
to examine in the intact rat the sequence of changes occurring in
a variety of physiological
parameters concerned with carbohydrate and lipid metabolism as a result of fasting aud subsequent
refeeding.
Groups of animals were deprived of food and refed
for the time periods indicated in t,hc legend t.o Fig. 1, after which
they were anesthetized and samples of plasma were analyzed for
free fatty acids, ketone bodies, glucose, and insulin.
In addition, the glycogen content of the livers was determined.
As shown in Fig. 1A little change took place in hepatic glycogen levels during the first 6 hours of starvation.
Thereafter,
however, the ccmcentration
fell precipitously
from a normal
value of approximately
50 mg per g of liver to 2.5 mg per g by
the 12-hour point.
No further change occurred between 12 and
24 hours, but at 48 hours the level increased to 10 mg per g.
This was followed by a decline after a further 2 days of fasting.
Refeeding rats that had been fasted for 24 hours resulted in a
rapid and approximately
linear rate of glycogen deposition in
the liver so that after 6 hours normal levels had been restored.
Although
much less pronounced,
changes in plasma glucose
concentration
followed a qualitatively
similar pattern to those
observed for liver glycogen as starvation progressed (Fig. 1C).
Again, no difference was noted between normal and B-hour fasting
values but between 6 and 12 hours the concentration
fell from
185 to 118 mg/lOO m1.l This level was maintained
over the
next 12 hours but gradually declined on more prolonged fasting
to a value of 95 mg/lOO ml after 96 hours.
Plasma insulin concentrations
were not measured in the individual animals of each experimental
group but from pooled
samples at each time point.
It can be seen that variations in
this parameter roughly paralleled those seen in plasma glucose
and liver glycogen levels (Fig. 1B). Refeeding resulted in the
expected brisk rise in plasma glucose and insulin concentrations,
both of which fell toward base-line values over a 4-hour period.
The plasma free fatty acid concentration
in normal animals
EJects

of Starvation

1 The initial high values for plasma glucose concentrations were

the consequence of the large carbohydrate
load administered at
the start of the experiment.

Downloaded from http://www.jbc.org/ by guest on August 25, 2016

EXPERIMENTAL

albumin-NaCl
solution containing
20 PCi of the labeled fatty
acid (specific activity, 56.3 PCi per pmole) was injected into the
circulating fluid after 30 min of perfusion.
Thirty minutes later
livers were rapidly frozen in liquid Nz and samples of perfusate
were taken for analysis.
Analyses on Perfusion Pluid and Liver-The
concentrat,ions of
pyruvate and lactate in the cell-free fluid were determined by
the procedures described for acetoacetate and P-hydroxybutyrate, respectively
(13), with the modification
that P-hydroxybutyrate dehydrogenase was replaced by lactate dehydrogenase
(EC 1.1.1.27).
In experiments where [3-14C]lactate was included in the perfusion fluid the incorporation
of isotope into
glucose and liver glycogen was determined using slight modifications of the procedures described by Eston and Park (14).
All other methodology
has been described in previous reports
(5, 12). In the text the term ketone bodies will always refer
to the sum of acetoacetate and fl-hydrosybutyrate.
illuteri&-Unlabeled
oleic acid was obt.ained from Sigma.
In all perfusion experiments the fatty acid was bound to the
bovine albumin (Fraction V, Armour Pharmaceutical
Co., Lot
G35808) by the method of Van Harken et al. (15). [1-14C]Oleic
acid and L-[3-%]lactate
were from new England Nuclear.

272
was about 0.25 peq per ml, but this value doubled
of starvation and continued to increase until the
at which time it had reached 0.8 peq per ml. The
remained at this level throughout
the following 36

slightly during the last 2 days of starvation (Fig. 1E). Despite

the increased mobilization
of free fatty acids in the first 6 hours,
plasma ketone levels at this point rose slightly but not significantly above nonfasting values (Fig. lo), confirming the now
well documented fact that increased delivery of free fatty acids
to the liver is not in itself sufficient to provoke a proportional
acceleration of hepatic ketogenesis (8). Between 6 and 12 hours,
however, plasma ketones increased from 28 to 181 pmoles/lOO
ml. Although the mean concentrations
at 24, 48, and 96 hours
were found to be 295, 155, and 201 pmoles/lOO ml, respectively,
these numbers were not statistically different from the value at
the la-hour point.
On refeeding, the starvation ketosis was
promptly reversed, with plasma ketone levels falling to normal
values after 30 min. At this point the plasma free fatty acid
concentration
had decreased from 0.8 to 0.4 peq per ml and after
1 hour had completely reverted to nonfasting values.

after 6 hours
12-hour point
concentration
hours but fell

E$ect

TABLE

acids

in medium

Livers were perfused in the manner described in the text. The

initial 80 ml of perfusion medium (hematocrit, 20%) contained
free fatty acids in a concentration
of approximately
0.7 mM in
the cell-free fluid.
A solution containing 12.5 mM oleate and 10%
albumin
throughout

in NaCl
the

State

Normal.
Fasted12
Fasted
Fasted
Fasted
Fasted
Fasted

solution
was infused
experimental
period.

of animal

at a rate
Within

of 75 pl per min
each
group
of

Additions

.
24
48
96
24
24

hr.......................
hr.......................
hr.
hr..
hr..
.
hr and refed

.
.
3 hr. .

(+)-Decanoylcarnitine

on Metabolism

in

during

perfusion

of rat hers

with

oleic

acid

experiments samples obtained at a given time point were mixed

in equal volumes and an aliquot of the mixture was assayed for
The data refer to the mean fatty
acid
free fatty acid content.
concentration
fusion
fluid

in microequivalents
per
from two to four experiments.

Minutes

ml

of

the

cell-free

per-

of perfusion

13

30

4.5

0.66
0.67
0.72
0.66
0.69
0.74
0.71

0.65
0.78
0.76
0.66
0.67
0.76
0.68

0.73
0.78
0.79
0.67
0.72
0.76
0.68

0.82
0.81
0.83
0.70
0.72
0.79
0.67

~^

60

0.85
0.85
0.82
0.74
0.74
0.85
0.76

Downloaded from http://www.jbc.org/ by guest on August 25, 2016

FIG. 1. Effects of fasting and refeeding

on metabolic parameters in the intact rat. Animals were fasted and refed for the
indicated time periods as described under Experimental
Procedure.
Each point represents the mean value f S.E.M. for
four or five animals.

of free fatty

and Refeeding
Perfused
Rat Liver

Ketogenesis-Previous
studies from this and other laboratories
have shown that after fasting periods of either 24 or 48 hours
the capacity of the isolated perfused rat liver to synthesize
ketone bodies from fatty acids and glucose from lactate is greatly
enhanced (2, 5, 16). Because of the rapid and pronounced
effects of starvation and refeeding on the metabolic parameters
studied in viva (Fig. 1) the possibility existed that similar kinet.ics
might be observed with regard to changes in fatty acid and
glucose metabolism in the isolated liver. To this end a second
batch of rats was subjected to periods of fasting and refeeding
identical with those used in the in viva experiments.
At the
appropriate
times the livers were perfused with oleic acid as
described under Experimental
Procedure.
Since our primary
interest during this investigation was in the effects of starvation
and refeeding on a hepatic fatty acid metabolism it was decided
to choose an initial concentration
of fatty acid close to that observed in the plasma of fasted rats and to maintain this level in
To
the perfusion fluid throughout the course of the experiment.
illustrate that these conditions were in general satisfied, representative examples showing the fatty acid concentration
in the
perfusion fluid at different time points are shown in Table I.
It is seen that, irrespective of the nutritional
state of the donor
animal, the uptake of oleate by the liver was approximately
the
same and remained fairly constant with time in all experimental

HOURS OF STARVATION

Concentration

of Starvation

273
-Starvation

200

8.
t

IO

,,

15 20
25
30 Gk+&
HOURS OF STARVATION

ments the perfusion medium contained

nondialyzed

erythrocyt.es.

The notation
glucose
production
- 35 lactate
uptake
(A)
should
be read glucose
production
minus
35 lactate
uptake.
Each point represents
the mean value
f S.E.M.
for determinations made after GO min of perfusion
with four to eight livers.
groups.
In addition, the presence of a potent inhibitor of long
chain fatty acid osidation, ( +)-decanoylcarnitine,
had no effect
on the uptake of oleate by livers from 24-hour fasted rats.
The data displayed in Fig. 2C illustrate the effects of starvation
and refeeding on ketone body production
in the perfused liver.
There is a striking qualitative similarity between the effects of
the same dietary manipulations
on ketone production
by the
isolated liver and plasma ketone levels in tivo (Fig. 10).
Thus,
little change took place in ketogenesis in the first 6 hours of
fasting.
The next 3 hours, however, were accompanied
by
clear-cut alterations in the metabolic profile of the liver.
During
this time ketone body synthesis over the 60-min perfusion period
increased sharply from 39 to 131 pmoles/lOO g of body weight.2
A maximal output of 147 pmoles/lOO g of body weight was
achieved at the 12-hour point and this was unchanged at 24
hours. Thereafter the rate declined somewhat such that at 48
and 96 hours values were in the region of 100 pmoles/lOO g of
body weight.
As was the case with starvation the effect of refeeding on ketone production
from oleate was also brisk, the
output falling from 147 to 80 ~moles/lOO g of body weight in the
first hour. By 3 hours the value was reduced to 36 pmoles/lOO
g of body weight, a rate which was not different from that exhibited by livers from nonfasted rats.
Gluconeogenesis
from Lactate-Preliminary
studies revealed
that the perfusion fluid as routinely prepared contained significant quantities of lactate (generally in the region of 3 mu).
Since this compound serves as an excellent gluconeogenic
substrate it was of interest to determine the effects of starvation
and refeeding on its uptake by the liver. These data are shown

2 As will be discussed later, rates of ketogenesis

in livers
from
fasted
rats tended
to increase
during
the second
half hour
of
perfusion.
While the data of Figs. 2C and 30 refer to ketone
body
production
over 60 min of perfusion,
qualitatively
identical
curves
are obtained
utilizing
data from the 30-min
time point,.

HOURS OF STARVATION

30

FIG. 3. Effects of fasting

and refeeding
on the metabolism
of
the isolated
perfused
rat liver.
All experiments
were carried
out
in a fashion
identical
with that described
in the legend to Fig. 2
with the exception
that L-[3-Wllactate
(2pCi)
was included
in the
perfusion
fluid
and the incorporation
of isotope
into perfusate
glucose
and liver glycogen
was determined.
The notation
glucose production
- 36 lactate
uptake
(A) should be read glucose
production
minus x lactate
uptake.
Each point represents
the
mean value f

S.E.M.

perfusion

four

with

for determinations

made after GO min of

livers.

in Fig. 2B. As was the case for ketone production, the rate of
lactate uptake increased between 6 and 9 hours of starvation
and plateaued between 12 and 24 hours, at which time an uptake
of 212 pmoles/lOO g of body weight was recorded.
After 48
hours this value had fallen to 177 and remained at this level
during a further 2 days of fasting.
It is noteworthy,
however,
that while the enhanced capacity of the liver to take up lactate
from the medium and to convert oleate into ketone bodies
appeared to correlate during starvation this was clearly not the
case in the reversal phase. Thus, 2 hours of refeeding, which
caused a 54% reduction in ketogenesis, had no significant effect
on lactate uptake.
In general, therefore, the rate of decline in
lactate uptake was considerably
slower than the reversal of
ketogenesis.
In order to determine whether the amount of lactate consumed
by the livers in different nutritional
states adequately reflected
relative rates of gluconeogenesis it was necessary to repeat these
experiments using perfusion fluid containing a known quantity
of labeled lactate and to measure the incorporation
of isotope into
glucose and liver glycogen.
The results of these studies are
shown in Fig. 3 from which it may be seen that the effects of

Downloaded from http://www.jbc.org/ by guest on August 25, 2016

FIG. 2. Effects of fasting

and refeeding
on the metabolism
of
the isolated
perfused
rat liver.
Animals
were fasted and refed for
the indicated
time periods
and livers were perfused
with oleic acid
as described under Experimental
Procedure.
In all experi-

274

3 It is recognized that the value given by t.he function glucose

production
- 55 lactate upt,ake will not provide an absoIute
measurement of hepatic glycogen content.
This is particularly
true in the case of livers from nonfasted animals or those that have
been fasted and refed, since in both cases only a portion of the
glycogen stores is released as glucose under the experimental
conditions used here. This point is well illustrated
by the fact
that such livers, when perfused in the presence of glucagon, will
increase their glucose output from 250 to 650 pmoles per 100 g
of body weight over a GO-min period (unpublished observations).
For this reason the data of Figs. 2A and 3A should be considered
only a reflection of the relative glycogen content of the liver under
different nutritional
conditions.

gested that the converse of t,he thesis might, more accurately

represent t.he actual situation, i.e. that the transport of t,he long
chain fatty acid from t,he cytoplasm into t.he mitochondrion
constitutes the primary control of the rate of such oxidat.ion and
that the rate of triglyceride synthesis is secondarily governed in
part by the activity of this fatty acid oxidation (5, 7, 8).
This problem has now been investigated in more detail using
techniques similar to those described in an earlier report (5).
However, two differences in methodology
should be noted.
First, whereas in the past we have routinely induced starvation
ketosis by fasting the rats for 48 hours, it now appears that a
24-hour fast is sufficient and this period has been used in all
subsequent experiments.
Second, unlike the earlier system in
which the specific activity of the labeled oleate in the perfusion
fluid was kept constant over the ent.ire 60-min period, in the
present experiments the label was int,roduced in trace quantities
as a single injection at the 30.min time point.
In this case,
then, the specific radioactivity
of the oleate declined during the
30- to 60-min interval because of the continuous infusion of unlabeled fatty acid into the pool. While the latter method was
adopted chiefly for its practical advantages it should be emphasized that direct comparison of the two techniques indicated that
both yielded essentially the same information.
For ease of presentation of the data the effects of various experimental
manipulations
on the time course of ketone body
synt,hesis, lactate uptake, and glucose production over t.he entire
perfusion period will be described first (Figs. 4 and 5). Thereafter, the disposal of labeled oleate betlveen 30 and 60 min in
these same experiments will be discussed.
As expected, it was observed that the la&ate initially
contributed to the perfusion fluid by the erythrocytes was removed
more rapidly by livers from fasted rats than by those frorn normal animals (controls, Fig. 4A).
Total lactate uptake after
60 min was 113 and 167 pmoles/lOO g of body weight in the
normal and fasted groups, respectively.
In the same time period
the values for glucose output were 235 and 107 pmoles/lOO g
of body weight (Fig. 4B). In the fasted group, therefore, the
quantity of lactate consumed could have accounted for 80%
of the glucose released, whereas in livers from normal animals
lactate uptake could have contributed at most only 257, of the
glucose produced.4
The much larger quantity of glucose released
in the latter case reflects the rapid rate of glycogenolysis commonly observed when such livers are perfused in vitro in the
absence of added glucose.
The kinetics of ketone body synthesis in these experiments is
shown in Fig. 4C. In the control livers (i.e. those perfused with
regular medium in which the red cells were not dialyzed and to
which no lactate was added) ketone production
after 60 min
was 24 and 141 pmoles/lOO g of body weight in livers frorn normal
It will be observed, however, that
and fasted rats, respectively.
under these conditions the rate of ketogcnesis in the fasted group
was nonlinear with time, the rates being 52 pmoles/lOO g of body
weight in the first half of the perfusion period and 89 pmolcs/lOO
g of body weight in the second $5 hour. Since lactate is known
to be strongly antiketogenic
with long chain fatty acids as substrate (5, 14), it seemed possible that the increased rate of ketogenesis in the second half of the perfusion period might have
been a consequence of a diminished lactate concentration in the
perfusate during this time. It was of interest, therefore, to
compare the rates of ketone production
and gluconeogenesis in
4 Throughout
these perfusion studies the quantity of pyruvate
released from the livers was negligible.
Data on this point have
therefore been omitted.

Downloaded from http://www.jbc.org/ by guest on August 25, 2016

starvation and refeeding on ketone production

(Fig. 30) and
lactate uptake (Fig. 3C) were essentially identical with those
observed in the previous experiment (Fig. 2, B and C). The
important point here, however, is that the incorporation
of isotope from 13.Vllnctate
into glucose released to the perfusate
and that stored as glycogen in the liver (the former invariably
representing 90 to 95/c of the total isotope incorporated)
paralleled almost exactly the kinetics fcr lactate uptake (cf. Fig. 3, B
and C). III addition, with the possible exception of the livers
from nonfasted animals, in all other experimental
groups the
calculated flux of lactate through the gluconeogcnic sequence as
determined by the isotope recovery in glucose and glycogen
accounted for the bulk (73 Do lOOo/,) of the lactate removed from
the perfusion fluid as measured enzymatically.
These findings
are in good agreemeut with the report by Exton and Park (14)
that lactate consumed by perfused livers from 24-hour fasted
rats is converted almost qua.ntitatively
into glucose.
Since glucose output from the liver may result from a combination of both glycogenolysis and gluconeogenesis, the former
being the major component in the early stages of fasting and the
latter the predominant
event at later time points, little inforrnation would be gained from a plot of glucose output versus time of
starvation.
However, since most of the lactate removed from
the medium was converted into glucose, then changes in the
value given by the number of micromoles of glucose produced
minus one-half of the micromoles of lactate consumed might be
espected to reflect variations in the glycogen content of the liver.
These calculations were made and are shown in Figs. 2A and
3A, from which a11 inverse correlation between these values and
those obtained for ketone production
and lactate uptake is
readily apparent.
A second noteworthy
feature of these data
is the virtually
identical configuration
of the curves obtained
from in vitro studies (Figs. 2A and 3A) with that showing the
absolute glycogen content of the livers in &JO (Fig. 1A). Under
the experimental
conditions used here, therefore, differences in
the amount of non-lactate-derived
glucose would appear to be a
reasonably good index of the relative state of glycogen stores in
the liver.3
EJect of Lactate on Ketogenesis and Glucose Production by Perfused Livers from Norlnal and 24-Hour Fasted Rats-Previous
studies in which livers from normal and 4%hour fasted rats were
perfused with [I-*C]oleic acid clearly demonst,rated that the
latter oxidized a larger fraction and esterified less of the fatty
acid than did the former (5). It was also established that a
variety of antiketogenic
compounds, including lactate, exerted
their effects primarily by causing a diversion of the fatty acid
from an oxidative fate into the pathway of triglyceride
and
While those data were interpreted
phospholipid
biosynthesis.
as being consistent with the widely held view that the fraction
of a long chain fatty acid load oxidized by the liver depends upon
the efficiency of its triglyceride-synthesizing
machinery,
we
pointed out that this concept might in fact be incorrect and sug-

275
0 Normal,
Fasted,
=Fasted,
AFasted,

Controls
Controls
Low Loctote
High Lactate

l Controls

15

30
45
MINUTES

60

FIG. 4. Effect of lactate concent,ration

on rates of ketogenesis
and glnconeogenesis in perfused livers from fasted rats. Livers
from normal and 24.hoIn fasted rats were perfused with oleic acid
as described lmder Experimental
Procedure.
Controls
refers to perfusions in which the erythrocytes were not dialyzed
to remove lactate.
Low lactate indicates that the red cells
High lactate denotes the addition of lactate to
were dialyzed.
medium containing lIondialyzed red cells. Results are given as
Four livers were perfused in each group
mean
values f P.E.N.
except the fasted controls where eight experiments were carried
out.

livers from fasted nninlals under conditions of low, intermediate,

and high concent8rations of lactate in the medium.
Ss illustrated
in Fig. 4 dialysis of the erythrocytes before use resulted in the
removal of most of the lactate from the perfusion fluid, an almost
complete cessat,ion of gluconeogenesis,
and a markedly stirnulated rate of ket)one production during the first 30 min of perfusion. Betweeu 30 and 60 min, however, rates of ketogenesis
were similar whether regular or lactate-depleted
cells were used.
When the initial lactate concentration
was set at 10 m;\l and
maintained at a high lrl-cl by constant infusion, ketone body
synthesis was greatly depressed to a rate that was considerably
lower than the controls with a shift of the curve towards linearity.
Under t)hese circumstances, and in agreement with previous observations (a), gluconcogeuesis was accelerated to very high and
probably maximal rates. It should be noted that n-addition
of high concentrations of lactate to perfusion fluid containing
dialyzed red cells resulted in rates of ketogenesis and gluconeogenesis identical with those obtained using nondialyzed
cells
These findings
in the presence of high concentrations of lactate.
indicate t,hat the stimulatory effect of dialysis was probably due
to the removal of lactate.
It was also observed that perfusion
of livers from normal animals with lactate-depleted
medium was

FIG.
5. Effect of (+)-decanoplcarnitine
on ketogenesis and
gluconeogenesis in perfused livers from fasted rats. Livers from
24.hour fasted rats were perfused with oleic acid in the presence
or absence of 0.5 mM (t^)-decanoylcarnitine
as described under
Experimental
Procedure.
The aerfusion fluid contained nondialyzed erythrocytes.
Results are given as mean values &
S.E.M. for eight control livers and four livers with (+)-decanoylcarnitine.

totally without effect on the rate of ketogenesis or glucose output.

Effect of (+)-Decanoylcarnitine
on Ketogenesis, Lactate Uptake,
and Glucose Output by Perfused Livers from d&Hour Fasted Rats(+)-Decanoylcarnitine
is a potent inhibitor
of long chain fatty
acid oxidation in animal tissues by virtue of its ability to inhibit
the activity of acylcarnitinetransferase
which is essential for the
transport of long chain fatty acids from the cytosol into the
mitochondrion
(17). The main reason for the use of (+)-decanoylcarnitine
in the present studies was to examine its effect
on the fate of oleic acid in livers from fasted rats. We were
particularly
interested to determine whether the presence of
this inhibitor would restore the pattern of fatty acid metabolism
in livers from fasted rats to that displayed by livers from normal
animals.
As can be seen from the data shown in Fig. 5C this
compound
caused an almost complete block in ketone body
production
from oleatc by livers from fasted rats. In esperiments not reported here it was shown, in agreement with other
investigators
(17), that octanoic acid, which does not require
the acylcarnitinetransferase
mechanism for its entry into the
mitochondrion
(17, 18), sustained high rates of ketogenesjs in
the prcscnce or absence of (+)-decanoylcarnitine.
While the curves shown in Fig. 5B might, at first glance, indicate a significant depression of gluconeogenesis by (+)-decanoylcarnitine, in actuality the small difference in glucose output
between the two experimental groups could be explained entirely
on the basis of the fact that the initial lactate concentration
in
the control group happened to be higher (3.41 mM) than that
(2.72 mM) in the (+)-decanoylcarnitine-treated
livers (Fig. 5A).
Since in this system the Km for lactate with regard to glucose
production is approximately
2 mM (14)) fluctuations in its initial
concentration
of the dimensions encountered in these studies

Downloaded from http://www.jbc.org/ by guest on August 25, 2016

MINUTES

276
In agreement with our earlier report (5) the dramatic stimulatory effect of starvation on total ketone body synthesis was
a.ccornpanied by a striking increase in the flow of label through
the ketogenic pathway and a marked decrease in its incorporation
into lipids (Lines 1 and 2, Table II).
Removal of lactate from
the medium by dialysis greatly stimulated
the initial rate of
ketogenesis (0 to 30 min) in livers from fasted rats while comparable rates were observed with both dialyzed and nondialyzed
cells over the 30- to 60.min interval (Fig. 4C). For this reason
it was to be expected that in the latter time period the distribution of oleate between oxidative and esterification
pathways
would be similar under both circumstances and, as shown in
Lines 2 and 3 of Table II, this was found to be the case. However, when high concentrations
of lactate were added its antiketogenic effect was accompanied by diversion of the fat.t,y acid
from the /3 oxidation sequence into the esterification
pat,hway
(Lines 2 and 4, Table II) in agreement with previous findings
(5). As mentioned above, removal of the lactate from the medium perfusing livers from normal animals had no effect on the
rate of ketogenesis at any time point in the perfusions.
Consistent with this finding were the essentially identical distribution
patterns of labeled oleate when these livers were perfused with
dialyzed or nondialyaed cells (Lines 1 and 5, Table II).
The most significant aspect of this series of experiments was
the finding that (+)-decanoylcarnitine,
which virtually
completely blocked fatty acid oxidation, had the effect of switching
acutely t,he mode of disposal of oleate in livers from fasted rats
to a pattern characteristic of livers from normal animals, i.e. in
the presence of the inhibitor
ketogenesis in livers from fasted
rats was reduced by 907, and virtually all of the fatty acid taken
up by the tissue was esterified (Lines 1,2, and 6, Table II).
This
observation
clearly demonstrates
that no fundamental
defect
exists in the esterifying capacity of the liver in the fasted state
and supports the view that the diminished ketogenesis and enhanced esterification
of oleate in livers from well fed animals
results primarily from a greatly depressed capacity to oxidize
long chain fatty acids. Moreover, this effect of (+)-decanoylcarnitine on the metabolism of oleic acid was exerted irrespective
of the lactate concentration in the perfusion fluid (Lines 6 and 7,
Table 11). This observation suggests that, under circumstances
where the ability to oxidize the fatty a.cid is blocked sufficient

TABLE

Metabolism

II

of [f-W]oleic acid in perfused livers from normal and E-hour fasted rats
fasted rats were perfused as at an initial
concentration
of 0.5 mM.
L-($)-Lactate

Livers
from normal and 24-hour
described
in the legends
to Figs. 3 apd 4. The concentration
of
fatty
acid in the cell-free
perfusate
was maintained
at approximately
0.75 mM by the constant
infusion
of albumin-bound
oleate.

When used, [1-%?]oleate

at

the

30-min

State of animal

time

point.

was added in trace quantities

(20 pCi)
(+)-D
ecanoylcarnitine
was present

Red cells used

Other

additions
pm&s/f00

1. Normal

6. Fasted

(4). .

Nondialyzed
Nondialyzed
Dialyzed
Nondialyzed
Dialyzed
Nondialyzed

7. Fasted

(2).

Dialyzed

2.
3.
4.
5.

(4).

Fasted (8).
Fasted (4).
Fasted (4).
Normal (3).

initially
to bring
its
infused
to maintain
given for the 30- to
percentage
of recovery
ber of livers shown

Lactate
(+)-Decanoylcarnitine
(+)-Decanoylcarnitine

15
89
85
53
18
8.5
12

f
f
f
f
f
f

Used

Kethes

2.0
3.6
7.5
5.0
3.2
0.9

was added
concentration
to 10 mM and was constantly
a concentration
above 7.5 mM.
Values
are
60-min
time interval
and are presented
as
of isotope
(means f S.E.M.)
for the numin parentheses.

Liver

70 [I-CJoleale/fOO

35.3
39.8
42.5
39.6
38.8
35.7

39.0

f
f
f
f
f
zk

2.6
1.1
0.5
3.1
1.8
1.1

1.4
15.6
18.7
7.5
2.3
0.9
/

1.0

f
f
f
f
f
f

0.3
1.1
4.3
0.7
0.5
0.06

YLipids

31.4
14.2
16.1
23.6
33.6
27.9
43.7

i
f
f
i
f
f

1.4
1.0
0.9
2.7
0.9
2.7

97.5
89.5
92.4
91.5
96.9
92.5
105.5

f
f
i
f
f
f

3.1
1.7
3.7
1.2
0.6
1.8

Downloaded from http://www.jbc.org/ by guest on August 25, 2016

might be expected to have noticeable effects on the rate of gluconeogenesis. Thus, at the lactate concentrations
used here,
which are within the physiological range, the virtually complete
shut down in fatty acid oxidation has had little, if any, effect on
the ability of the liver to convert this substrate into glucose.
These findings appeared to contrast with those reported by
Williamson et al. (17) who showed that inhibition
of fatty acid
oxidation by ( +)-decanoylcarnitine
drastically
curtailed
the
rate of gluconeogenesis from lactate in the perfused rat liver.
Since these workers used much higher concentrations
of the
gluconeogenic substrate (8 to 12 mM) it was felt worthwhile
to
repeat our studies under conditions where the initial lactate
concentration was set at 10 InM and maintained at a high level by
constant infusion throughout
the perfusion period.
In experiments not shown ketogenesis was again virtually abolished by
the inhibitor, and over the 15- to 60-min interval glucose output
was reduced from 168 f 4.7 to 69 i 9.5 ~moles/lOO g of body
weight (means + S.E.M. of four perfusions in each group),
confirming the findings of Williamson el al. (17). It is apparent,
therefore, that fatty acid osidation is required for high but not
low rates of gluconeogenesis from lactate.
Metabolism of [I-14C]Oleic Acid in Perfused Livers from Normal
and %$-Hour Fasted Rats-As
mentioned under Experimental
Procedure, labeled oleate was added to the perfusion fluid at
the 30-min time point in the studies shown in Figs. 4 and 5. At
60 min the distribution
of the label between ketone bodies and
liver lipids was determined.
From the data compiled in Table
II, it is apparent that the quantity of isotope taken up by the
livers was approximately
the same in all experimental
groups
and was generally in the region of 40% of the amount added per
100 g of body weight.
This finding is consistent with other
reports on fatty acid metabolism in the perfused rat liver (5, 15,
19) and was also to be expected on the basis of the data shown
in Table I.
As noted previously (5), under the experimental
conditions
used here virtually all of the de novo-synthesized lipids (triglycerides and phospholipids)
were retained in the liver over the
time period studied.
The sum of the isotope recovered in lipids
and ketone bodies invariably accounted for the bulk of the fatty
By comparison, the amount of labeled oleate
a.cid metabolized.
converted into 14C02 was small and was not routinely measured.

277
ol-glycerophosphate
is supplied for its esterification
enous sources in the absence of an added precursor
phosphate.

from endogof cY-glycero-

DISCUSSION

Downloaded from http://www.jbc.org/ by guest on August 25, 2016

It is well established that livers taken from rats that have

been fasted for periods of 24 to 48 hours exhibit a markedly increased capacity to synthesize ketone bodies and glucose when
We
perfused with fatty acids or gluconeogenic precursors (l-5).
were interested to know whether the findings observed in the
isolated liver adequately reflected changes in physiological
parameters occurring in viva. In addition, since reported rates of
ketogenesis and gluconeogenesis appeared to be similar whether
24- or 48.hour fasted rats were used, we wished to determine the
time required for the full induction of both metabolic pathways.
Unexpectedly, all of the hallmarks of the ketotic state commonly observed on more prolonged fasting were fully developed
Thus at this time liver glycoafter only 12 hours of starvation.
gen stores were virtually exhausted, plasma glucose and insulin
concentrations were markedly diminished, and levels of plasma
free fatty acids and ketone bodies had attained high values.
Even more surprising was the finding that most of these changes
occurred during the brief period between 6 and 9 hours of starvation.
As expected on the basis of previous studies (20) the refeeding
of fasted animals resulted in a prompt reversal of the ketotic
state. This was accompanied by a sharp return to normal levels
of plasma free fatty acids, glucose, and insulin and a gradual
replenishment
of hepatic glycogen stores that was complete
after 6 hours.
We next carried out a series of esperiments with perfused
livers from animals treated in a manner identical with that used
in the in tivo studies. A striking synchrony was observed between the events taking place in the intact animals and the rates
of lactate uptake, gluconeogenesis,
glycogenolysis,
and ketogenesis found in the isolated livers during the development of
starvation ketosis (Figs. 1 to 3). While the uptake of oleate
from the perfusion fluid was approximately
constant in all experimental
groups, its conversion into ketone bodies had not
This finding
increased significantly after 6 hours of starvation.
was consistent with the in viva situation in which the concentration of plasma ketone bodies failed to increase during the first
6 hours despite the fact that plasma free fatty acids levels had
doubled in this time. The lack of correlation between plasma
free fatty acid concentration
and hepatic ketogenesis has been
commented on previously (5, 8, 12).
The concomitant enhancement in both ketogenesis from oleate
and gluconeogenesis from lactate observed in perfused livers
during the 6- to 12.hour fasting period might be taken as supportive evidence for the view that efficient operation of the gluconeogenic sequence is dependent upon fatty acid oxidation (17,
21, 22). However, it should be noted that in several instances
a divergence between rates of gluconeogenesis and ketogenesis
was observed. First, with low concentrations of lactate (in the
region of 3 mM) livers from fasted rats synthesized glucose
at similar rates whether fatty acid oxidation and ketone body
formation were active or totally blocked with (+)-decanoylcarnitine (Fig. 5). In contrast, with high levels of lactate (approximately 10 mM), sufficient to promote maximal rates of gluconeogenesis, the inhibition of fatty acid oxidation by (+)-decanoylcarnitine
greatly decreased glucose output.
Second, during
the early stages of reversal of starvation ketosis by refeeding
(Figs. 2 and 3) ketogenesis was inhibited, while lactate uptake

and gluconeogenesis were unaffected.

Third, when livers from
normal animals were perfused with octanoic acid (experiments
not shown), high rates of ketogenesis were obtained without
stimulation of lactate uptake, in agreement with the findings of
Exton and Park (14). These observations indicate that the
relationship
between fatty acid oxidation and gluconeogenesis
is not a simple one. The initial activation of the gluconeogenic
sequence by fasting is probably independent
of fatty acid oxidation.
Once initiated, the capacity of the system to convert
lactate into glucose is markedly elevated but seems to require
fatty acid oxidation only when higher levels of this substrate
are used.
The primary purpose of the present investigation
was to examine the relationship
between fatty acid oxidation and esterification in the over-all regulation of ketogenesis.
A conventional
view of this relationship
would hold that the rate of fatty acid
oxidation in liver is regulated by the efficiency of the triglyceridesynthesizing system which in turn is governed by the availability
of oc-glycerophosphate
(6, 14). We have questioned this view
on the basis of the following observations: (a) total tissue content
of a-glycerophosphate
is actually higher in livers from fasted
rats than in those from normal animals (5) ; (5) no correlation
was seen between the capacity of various compounds to reverse
starvation ketosis in the rat and their effects on liver ol-glycerophosphate concentrations
(5, 9) ; and (c) livers from severely
ketotic diabetic rats are frequently observed to be engorged with
fat (7, 10, 23), a finding that is difficult to reconcile with the
viewpoint that impaired triglyceride synthesis plays a necessary
role in the induction of the ketotic state.
Consistent with our previous report (5) it was observed in
these experiments that when perfused with labeled oleate, livers
from normal and fasted rats extracted similar quantities of the
fatty acid, but, whereas the former group esterified virtually all
of the substrate, the latter esterified less than half, the rest being
converted into oxidative products, principally
ketone bodies.
The critical observation of these studies, however, was that
when (+)-decanoylcarnitine
was used to block fatty acid oxidation in livers from fasted animals, the oleic acid taken up was
now totally esterified.
In other words the effect of this maneuver
was to convert acutely the pattern of fatty acid metabolism of
the ketotic liver into that expected of normal hepatic tissue.
Moreover, this effect of (+)-decanoylcarnitine
could be demonstrated in the absence of any added precursor of Lu-glycerophosphate.
It is clear, therefore, that no fundamental defect exists
in the triglyceride-synthesizing
machinery of the liver in the
fasting state.
On the basis of these studies it is tempting to speculate that
the striking enhancement
of ketone body production
induced
by starvation is due primarily to an activation in the oxidative
sequence for long chain fatty acids. While the data allow no
firm conclusion on the precise point of regulation, it seems unlikely that the fi oxidation pathway itself is involved since octanoic acid, which enters the mitochondrion
by a carnitineindependent
mechanism, is oxidized at similar rates in livers
from normal and ketotic rats (12). The data would be consistent with the thesis that the control site for both fatty acid
oxidation and ketogenesis is at the acylcarnitinetransferase
reaction which presumably is the initial step in the oxidation sequence for long chain fatty acids (18, 24, 25). In this regard
it is noteworthy
that Norum (26) has reported an increased activity of this enzyme in homogenates of livers from ket.otic rats.
While the changes reported in that study were not of sufficient
magnitude to account for the differences in fatty acid oxidation

27s
observed herr, the possibility of metabolic regulation at this site
deserves further exploration
since measurements of enzyme activity in brokeu cell preparations
may not always reflect differences prevailing iu the intact organ (27-29).
Finally, if hepatic fatty acid metabolism is controlled primarily
by activation-inact,iration
of the acylcarnitinetransferase
reaction
it is intriguing
to ask how this is related to the simultaneous
changes seen here in other physioIogica1 parameters occurring
with fast)ing and refeeding.
Since lactate exerts its antiketogenic
effect by causing a diversion of long chain fatty acids from an
osidat.ive fate int.0 the t,riglyceride-synthesizing
pathway in the
absence of significant
changes in ol-glycerophosphate
levels,
further investigation of the mechanism of action of lactate and
other antiketogenica agents (5) may provide a clue to the means
whereby dietary and hormonal control of hepatic fatty acid
metabolism and ketogenesis is accomplished.

REFERENCES
1. HE&IS, I<., Ross, B. D., BIGRRY, M. N., BND KREHS, H. A.
(1966) I?kh.e~~~. J. 101, 284
2. EXTON, J. H., CO~BIN, J. G., AND PAINT, C. R. (1969) J. LGol.
Chem.

244,

4095

3. WILLIAMSON,
J. R., BROWNING,
J. Hiol. Ch,em. 244, 4607
4. MENAHAN,
L. A., AND WINAND,

E. T.,
0.

AND SCHOLZ,

(1969) Eur.

R. (19G9)

J. Biochem.

9,

182
5.

MCGARRY,

J.

D.,

AND

FOSTER,

D. W. (1971) J. Biol.

Chem.

6247
6. May~s,

P. A., ANU FELTS,

J. M. (1967) ~Vature 216, 716

246,

8.
9.
10.

MEIER,

J.

H., .4ND
MCGARRY,
WILLIAMSON,
KREBS,
CHERNIX,

J. D., FALOONA,
G. R., UNGEI~,
R.
W: (1972) J. Lipid Res. 13; 228
J. D., AND FOSTER, Il. W. (1972) Metabolism
21,471
D. H., VICLOSO, D., ELLINGTON,
E. V., L4~~
H. A. (1969) Biochem.
J. 114, 575
S. S., AND Scow, 1-L 0. (1959)
Amer. J. Physiol.
196,
M., MCGARRY,
FOSTER,
D.

125
11.

GOOD,
Chem.

C. A., KR~ME:R, H., AND SOMOGYI,

M. (1933) J. Biol.
100, 485
12. MCGARRY,
J. D., AND FO~TICR, D. W. (1971) J. Biol. Chem. 246,
1149
13. MCGAR~Y,
J. D., GUEST, M. J., AND FOSTER,
1~. W. (1970)
14.

EXTON,

J. Biol.

Chem.

J. H.,

2622
15. VAN HARKEN,
J. Biol.

16.
17.

245,
.~ND

11.

Chem.

R.,

4382
PARK,
DIXON,

(1967) J. Biol.

C. W.,

AND

HEIMB~:RG,

Chem.

242,

M. (1969)

244, 2278

H. A., WALLACE,
P. G., HEMS, R., AND
R. A. (1969) Riochem.
J. 112, 595
WILLIAMSON,
J. R., BROWNING,
E. T., SCHOLZ,
R.,
R. A., lick
FRITZ,
I. B. (1968) Diabetes 17. 194

FRITZ,
i. B. (19Gl) khysiol:
Rev: 41, 52
MORRIS,
B. (1963) J. Physiol.
168, 5G4
FOSTER,
D. W. (1967) J. C&in. Invest. 46, 1283
KREUS,

18.
19.
20.
21. SBLING,

H. D., WILLMS,
B.,
J. (1968) Eur. J. Biochem.

22.

C. R.

FRIEDRICHS,
4, 364

D.,

FK.EEDL.AND,
KREISI~ERG,

AND KLICINIXE,

J. R., BROWNING,
E. T., THURMAN,
R. G., AND
SCHOLZ, K. (1969) J. Biol. Chem. 244, 5055
23. HEIMBERG,
M., V.~N HaRKEN,
D. R., AND BROWN,
T. 0. (1967)
Biochim.
Biophys.
Acta 137, 435
24. FRITZ, I. B., BND Ma~auIs,
N. R. (1965) Proc. iVat. Acud. Sci.
U. 23. A. 54, 1228
25. AUGENFELD,
J., AND FRITZ, I. B. (1970) Can. J. Biochem.
48,
288
K. R. (1965) Biochim.
Biophys.
Acta 98, 652
26. NORUM,
27. WILLuvsoN,
I>. H., BATES, M. W., ANII KIUG~S, H. A. (1968)
Biochem.
J. 108, 353
28. KREBS,
H. A. (1967) 3. Konferenz
Gesellschaft
Biologische
Chemie, p. 129, Springer-Verlag,
Berlin, Heidelberg,
and
New York
J. I)., AND FOSTXR,
D. W. (1969) Biochim.
Biophys.
29. MCGARRY,
Acta 177, 35
WILLIAMSON,

Downloaded from http://www.jbc.org/ by guest on August 25, 2016

Acknow2edgnlclzts-\e
acknowledge with pleasure the expert
technical assistance of Ms. Petra Contreras and Ms. Martha
Cook. We are also iudebted to Dr. I. 13. Fritz and to Dr. Y.
Kawashima
(Otsuka Pharmaceutical
Factory,
Naruto,
ToInsulin
kushima, Japan) for gifts of ( +)-decanoylcarnitine.
assays were generously performed by the laboratory of Dr. Roger
H. Unger.

7.

The Effects of Starvation and Refeeding on Carbohydrate and Lipid Metabolism in

Vivo and in the Perfused Rat Liver : THE RELATIONSHIP BETWEEN FATTY
ACID OXIDATION AND ESTERIFICATION IN THE REGULATION OF
KETOGENESIS REGULATION OF KETOGENESIS
J. Denis McGarry, Jrgen M. Meier and Daniel W. Foster
J. Biol. Chem. 1973, 248:270-278.

Access the most updated version of this article at http://www.jbc.org/content/248/1/270

Alerts:
When this article is cited
When a correction for this article is posted

This article cites 0 references, 0 of which can be accessed free at

http://www.jbc.org/content/248/1/270.full.html#ref-list-1

Downloaded from http://www.jbc.org/ by guest on August 25, 2016

Click here to choose from all of JBC's e-mail alerts