299
Effects of food restriction on the responses of the mammary gland
and adipose tissue to prolactin and growth hormone in the
lactating rat
D J Flint and R G Vernon
Hannah Research Institute, Ayr KA6 5HL, UK
(Requests for offprints should be addressed to D J Flint)
Abstract
Exogenous GH is used extensively in the USA to stimulate
milk production in dairy cattle but its eVectiveness is
reduced in undernourished animals. It has been proposed
that GH increases milk yield by stimulating IGF-I secre-
tion and that this IGF-I-response is nutritionally sensitive
and thus acts as a ‘sensor’ of energy balance. To investigate
this possibility, we placed lactating rats on three planes of
nutrition, ad libitum, 50% or 25% of ad libitum for 48 h.
Subgroups of these animals were treated for 48 h with
bromocriptine, to suppress prolactin secretion, and anti-rat
GH, to neutralize GH action. From 24 to 48 h some of the
treated animals were assessed for their milk yield response
to prolactin or GH.
Food restriction reduced milk yield in control rats by
approximately 50% and was accompanied by a catabolic
state, as judged by lipid mobilization from adipose tissue
and by low concentrations of serum insulin, IGF-I, tri-
iodothyronine and thyroxine, and increased serum non-
esterified fatty acid concentrations. In animals fed ad
libitum, anti-rat GH plus bromocriptine treatment pro-
duced an 80% decrease in milk yield and a dramatic fall in
the activity of acetyl-CoA carboxylase in mammary tissue.
GH was able to stimulate milk yield when given from 24
to 48 h; however, its eVectiveness decreased progress-
ively as food intake was reduced. The milk yield
response to GH was accompanied by an increase in serum
IGF-I concentrations and this response also decreased
Introduction
Growth hormone (GH) stimulates milk production during
lactation and it has been proposed that this is mediated
indirectly via the production of insulin-like growth
factor-I (IGF-I) (see Bauman & Vernon 1993). Use of
exogenous GH to stimulate milk yield is now common in
the USA, and concern has been expressed that this may
‘push’ animals into negative energy balance and run the
risk of ‘metabolic collapse’. When nutrition is inadequate,
however, the eVect of GH on milk output is considerably
Journal of Endocrinology (1998) 156, 299–305 ? 1998 Journal of Endocrinology Ltd Printed in Great Britain
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progressively with reduction of food intake, consistent
with the hypothesis that IGF-I determines the milk yield
response to GH and thus regulates GH action on the
mammary gland in a nutritionally dependent fashion.
However, the milk yield response to prolactin and the
milk yield of control rats decreased in line with food intake
without any changes in serum IGF-I concentrations. This
clearly indicates that factors other than IGF-I are respon-
sible for restricting milk yield. In order to assess other
possible candidates for this role, we monitored serum
glucose, non-esterified fatty acids, insulin triiodo-
thyronine and thyroxine concentrations, but found no
evidence for any simple relationship between these par-
ameters and the milk yield response to prolactin and GH.
Surprisingly we found that the ability of GH or prolactin
to prevent epithelial cell loss in the mammary gland was
completely insensitive to nutrient intake, despite the fact
that IGF-I is considered to be an important survival factor
for mammary epithelial cells.
Finally, we also demonstrated that, at least during
short-term food restriction, the lactating rat is capable of
mobilizing significant amounts of lipid from adipose tissue,
such that it could provide the total output of triglyceride in
milk, which is much greater than has previously been
proposed.
Journal of Endocrinology (1998) 156, 299–305
reduced and, consistent with this, so is the increase in
serum IGF-I concentrations (McGuire et al. 1992, 1995).
It has thus been proposed that IGF-I serves as an indicator
of ‘energy balance’ (McGuire et al. 1995) and thus limits
any potentially harmful eVects of exogenous GH. Consist-
ent with this hypothesis, serum concentrations of IGF-I
are characteristically high when animals are in positive
energy balance and low when they are in negative energy
balance.
In order to test the hypothesis that IGF-I is a sensor of
energy balance for milk production in the lactating rat, we
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300 D J FLINT and R G VERNON · Food restriction and the milk yield response to prolactin and GH
took advantage of a model we have developed in the
lactating rat in which both GH and prolactin (PRL)
stimulate milk secretion (Barber et al. 1992, Flint et al.
1992). Since the eVects of exogenous GH and PRL are
only evident in GH- and PRL-deficient rats, all animals
were first treated with anti-rat (r)GH and bromocriptine.
Whereas the eVects of GH may be mediated via IGF-I,
there is no evidence that the metabolic eVects of PRL also
require IGF-I to serve as a mediator for its actions on the
mammary gland. We therefore examined the influence
of food restriction on the response to GH and PRL
to examine its eVects on milk secretion stimulated via
IGF-I-dependent and -independent mechanisms.
To assess the metabolic status of the animals we
measured several parameters related to adipose tissue
metabolism, since animals in negative energy balance have
to mobilize lipid from adipose tissue whereas, when in
positive energy balance, they store excess energy in
adipose tissue. Adipocyte mean cell volume was measured,
as this is related to loss or gain of lipid, serum non-
esterified fatty acids (NEFAs) were determined as indi-
cators of lipolysis, and acetyl-CoA carboxylase activity was
assayed as an indicator of lipogenesis (Vernon 1980). A
number of other parameters were also measured as possible
signals of energy status, including glucose, insulin and
thyroid hormones.
Materials and Methods
Animals
Female Wistar rats, approximately 250 g body weight,
were mated and, at parturition, litters were adjusted to
10 pups. All animals were allowed free access to water
and food (Labsure irradiated CRM diet, Labsure, Poole,
Dorset, UK) except as described below.
Hormones
Recombinant bovine GH (bGH) was a gift from
Monsanto (St Louis, MO, USA), and ovine (o)PRL-20
was a gift from NIDDK, Bethesda, MD, USA.
Preparation of sheep anti-rGH serum
Sheep were immunized with rGH and the antiserum was
assessed for specificity as previously described (Madon et al.
1986). This antiserum has previously been shown to
suppress body weight gain and serum IGF-I concen-
trations by 90% in rapidly growing rats at the doses used in
this study.
Experimental protocol
Lactating rats were treated, commencing on days 12–14 of
lactation, with a combination of bromocriptine (Sigma
Journal of Endocrinology (1998) 156, 299–305
Chemical Co, Poole, Dorset, UK; 500 µg/injection), to
suppress PRL secretion, and sheep anti-rGH „-globulin
(150 mg/injection), to neutralize rGH. Treatment was
administered twice daily for 2 days by s.c. injection.
During these 2 days, one group of the animals (H) were
allowed ad libitum access to food; another group had their
food intake restricted to 50% of their intake over the
previous 24 h (M) and a third group had their food intake
restricted to 25% of intake over the previous 24 h (L).
During the period 24–48 h, each group was further
subdivided into three groups. One received oPRL
(500 µg/injection), and another received bGH (500 µg/
injection). These doses have previously been shown to
completely prevent the decline in milk yield induced by
bromocriptine or anti-rGH treatment respectively. A third
group received excipient (0·1 M NaHCO3) at 1000 and
1700 h. An additional three groups of animals served as
untreated controls fed at H, M or L levels for the 48 h
period. Dam and litter weights were determined daily, and
food and water intakes were also recorded daily. Milk yield
was determined by the method of Sampson & Jansen
(1984). All animals were killed by cervical dislocation 48 h
after the start of treatment (i.e. 24 h after receiving PRL or
GH replacement therapy). Blood was obtained from the
trunk, and serum was prepared by centrifugation at
2000 g for 10 min and stored at "20 )C until analysed
for insulin (Vernon et al. 1981), IGF-I (Flint & Gardner
1989), tri-iodothyronine (T3) and thyroxine (T4) by RIA
(IDS, Boldon, Tyne and Wear, UK) and for serum glucose
(Bergmeyer et al. 1974) and NEFAs (Itaya & Ui 1965).
The right fourth, fifth and sixth abdominal inguinal
mammary glands were removed, weighed and frozen
in liquid N2 for determination of DNA concentration
(Labarca & Paigen 1980) and total acetyl-CoA carboxylase
activity, i.e. enzyme activity was measured after preincu-
bation with citrate and Mg2+ to activate any enzyme in
the inactive state (Borland et al. 1994). Parametrial adipose
tissue was also removed for determination of mean adi-
pocyte volume and total acetyl-CoA carboxylase activity
(Borland et al. 1994). Statistical analyses were performed
using ANOVA (general linearized model). Where statis-
tically significant eVects of plane of nutrition or hormone
treatment were observed, individual group means were
compared using Student’s t-test.
Results
Mammary gland metabolism
Milk yield was markedly suppressed by treatment for 48 h
with anti-rGH and bromocriptine (Fig. 1a). In ad libitum
fed rats, PRL or GH treatment from 24 to 48 h was able
to stimulate milk production although not back to control
values. This is due to the short treatment period of 24 h
since 2–3 days of PRL treatment can normalize milk
yield, whereas GH is slightly less eVective. The milk yield
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 Food restriction and the milk yield response to prolactin and GH
achieved in response to PRL or GH also decreased
progressively as food intake was reduced, as indeed did the
milk yield of the untreated lactating controls. Changes in
acetyl-CoA carboxylase activity (thought to be the rate-
limiting enzyme for fatty acid synthesis) in mammary
tissue paralleled changes in milk yield, with a progressive
loss of the ability of GH (P<0·01) or PRL (P<0·01) to
increase its concentration as food intake was restricted
(Fig. 1b). Mammary DNA content was reduced by
approximately 30% after treatment with anti-rGH and
bromocriptine (P<0·01). In striking contrast with the
metabolic eVects, both GH and PRL prevented this loss of
DNA, entirely independently of the levels of food intake
(Fig. 1c).
Serum IGF-I concentrations
Serum IGF-I concentrations were lower in ad libitum fed
lactating rats (31·7&3·2 nmol/l; mean&s.e.m., n=5) when
compared with non-lactating rats (63·6&4·3 nmol/l,
n=8; P<0·05). IGF-I concentrations decreased further
(P<0·01) when lactating rats were given anti-rGH and
bromocriptine, presumably because of the absence of
GH activity (Table 1). When GH was administered from
24 to 48 h, it induced a large increase in serum IGF-I
concentrations; however, this eVect of GH decreased
progressively as food intake was restricted (P<0·01). By
contrast, PRL did not influence serum IGF-I concen-
trations significantly.
Adipose tissue metabolism
When food intake was restricted to 50% of ad libitum there
were no significant eVects on the mean adipocyte volume
(Table 2). However, when food intake was restricted to
25% of ad libitum, there were significant decreases
(P<0·01) in adipocyte volume, indicative of major lipid
mobilization in all three groups of rats that were making
significant amounts of milk (i.e. control, PRL- and GH-
replacement groups). In the anti-rGH and bromocriptine-
treated animals, where milk yield was extremely low,
there was no evidence of significant loss of lipid from
adipose tissue with the 50% or 25% diets. Changes in the
activity of acetyl-CoA carboxylase in adipose tissue were
consistent with perceived energy status; thus, in all treat-
ment groups, reducing food intake decreased the activity
of this enzyme (Table 2). Conversely, decreasing milk
production by endocrine manipulation increased the
activity of acetyl-CoA carboxylase in adipose tissue. Thus
the endocrine manipulations used in this study produced
reciprocal changes in enzyme activity in mammary gland
and adipose tissue, whereas restricting food intake
decreased activity in both tissues.
Serum hormones and metabolites
Serum insulin concentrations increased 3-fold in anti-
rGH- and bromocriptine-treated rats compared with
· D J FLINT and R G VERNON 301
control lactating rats (P<0·01) (Table 1), again consistent
with a return to a positive energy balance in these animals.
PRL, but not GH, was able partially to prevent this
increase (P<0·01). Restriction of food intake led to a
progressive decline in serum insulin concentrations
(P<0·01) in all but the control lactating rats, where insulin
concentrations were already very low. Serum NEFA levels
increased in all groups when food intake was restricted
to 50% of ad libitum (1·1&0·1 and 1·7&0·1 mmol/l
(mean&s.e.m., n=20) for ad libitum fed and 50% food
intake respectively; P<0·01) but did not increase further
when food intake was restricted to 25% of ad libitum
(1·6&0·2 mmol/l). Serum T4 levels were reduced in
control lactating rats (31&4 nmol/l (mean&s.e.m., n=5))
compared with non-lactating animals (40&3 nmol/l,
n=5, P<0·05 Student’s t-test). T4 levels were not signifi-
cantly aVected by restricting food intake to 50% of ad
libitum but they did decline when intake was restricted to
25% of ad libitum (P<0·02, ANOVA) in control lactating
rats (22&1 nmol/l) or rats receiving PRL- (26&3 nmol/l)
or GH- (25&1 nmol/l) replacement therapy. By contrast,
T4 levels were increased (P<0·05) in rats given anti-rGH
and bromocriptine (41&2 nmol/l), such that they were
similar to values in non-lactating rats and were not
suppressed even when food intake was reduced to 25% of
ad libitum (42&7 nmol/l). Similar eVects were evident for
T3, with lactating animals exhibiting lower T3 levels than
non-lactating animals (1·15&0·07 vs 1·42&0·10 nmol/l,
P<0·05) and were increased in rats receiving anti-rGH
and bromocriptine treatment (1·61&0·2 nmol/l). Serum
T3 concentrations tended to decline with increasing food
restriction, although this did not achieve statistical signifi-
cance, and neither were T3:T4 ratios influenced signifi-
cantly (results not shown). Lactating rats exhibit a modest
hypoglycaemia compared with non-lactating animals.
Treatment with anti-rGH and bromocriptine, which re-
duced milk yield, produced a small increase in serum
glucose values (P<0·01), whereas PRL- but not GH-
replacement therapy produced a small decrease (P<0·05)
in serum glucose concentrations. The level of food intake,
however, failed to influence serum glucose in any of the
groups (results not shown).
Discussion
A number of studies have examined the eVects of under-
nutrition upon milk production in the rat, although many
have included food restriction during pregnancy (see
Rasmussen (1992) for review). However, three studies
have examined the eVects of food restriction solely during
lactation (Sainz et al. 1986, Taylor et al. 1986, Grigor et al.
1987), and the magnitude of the eVects on milk produc-
tion were very similar to those reported here. We have
extended their findings by demonstrating the ability of
rats, at least in the short term, to continue lactating at milk
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302 D J FLINT and R G VERNON · Food restriction and the milk yield response to prolactin and GH

Figure 1 Milk yield in ml/day (a), acetyl-CoA carboxylase activity (µmol/min per g tissue) in mammary tissue (b)
and DNA content of the mammary gland (c) in control lactating rats, lactating rats treated for 48 h with anti-rGH
plus bromocryptine without or with prolactin or GH from 24 to 48 h. Values are means& S.E.M. for five animals
per group. Results were analysed by ANOVA. Where significant differences were found, individual means were
tested using Student’s t-test. Values not sharing the same superscript differ (P<0·05).

Journal of Endocrinology (1998) 156, 299–305

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 Food restriction and the milk yield response to prolactin and GH · D J FLINT and R G VERNON 303

Table 1 Serum concentrations of IGF-I and insulin in lactating rats treated with anti-rGH and bromocriptine, with or without replacement
therapy with PRL or GH, subjected to three different levels of food intake. Values are means& S.E.M. for five animals per group

Ad libitum 50% food intake 25% food intake

IGF-I (nmol/l) Insulin (nmol/l) IGF-I (nmol/l) Insulin (nmol/l) IGF-I (nmol/l) Insulin (nmol/l)
a a a ac a
Control 31·7&3·2 0·10&0·03 24·4&2·1 0·12&0·04 22·9&1·7 0·10&0·01a
Anti-rGH and bromocriptine 18·3&2·8b 0·30&0·02b 16·1&2·5ab 0·18&0·02ac 15·5&1·9ab 0·12&0·02ac
Anti-rGH, bromocriptine and PRL 19·9&3·5b 0·19&0·03c 18·4&2·8ab 0·12&0·02a 15·3&0·07ab 0·06&0·02d
Anti-rGH, bromocriptine and bGH 70·5&5·5c 0·29&0·02b 54·3&4·1d 0·14&0·01ac 44·9&2·5e 0·10&0·03a

Results were analysed by ANOVA. Where significant differences were found, individual means were tested using Student’s t-test. Values not sharing the same
superscript within a parameter differ (P<0·05).

Table 2 Mean adipocyte volume and total acetyl-CoA carboxylase activity in adipose tissue in lactating rats treated with anti-rGH and
bromocriptine with or without replacement therapy with PRL or GH subjected to three different levels of food intake. Values are
means& S.E.M. for five animals per group

Ad libitum 50% food intake 25% food intake

Acetyl-CoA Acetyl-CoA Acetyl-CoA
carboxylase carboxylase carboxylase
Adipocyte (nmol/min Adipocyte (nmol/min Adipocyte (nmol/min
volume (pl) per g tissue) volume (pl) per g tissue) volume (pl) per g tissue)
Control 346&59a 64&25a 330&34a 32&18c 222&8b 8&4e
Anti-rGH and bromocriptine 381&50a 205&21b 405&57a 124&31d 359&54a 63&4c
Anti-rGH, bromocriptine and PRL 409&38a 203&23b 390&25a 96&17d 251&25b 40&10c
Anti-rGH, bromocriptine and bGH 316&24a 113&29ad 287&44a 71&14ad 249&46b 37&20c

Results were analysed by ANOVA. Where significant differences were found, individual means were tested using Student’s t-test. Values not sharing the same
superscript within a parameter differ (P<0·05).

yields in excess of 40% of normal, even when food intake
was reduced to 25% of ad libitum. During severe food
restriction, adipocyte mean cell volume fell from 346 pl to
222 pl (i.e. by 36%). Based upon a body fat content of 30 g
in our rats (Kanto & Clawson 1980), this would represent
a loss of approximately 11 g of lipid over 2 days. Milk yield
in our rats was 45 ml/day, with a milk fat concentration of
approximately 10% (Flint & Gardner 1994), representing a
milk fat output of 9 g in 2 days. Thus the mobilization of
adipose tissue could in theory provide all of the triglyceride
in milk. This suggests that adipose tissue can make an
important contribution to milk yield in rats, quantitatively
much greater than proposed by Sadurskis et al. (1991).
Clearly, however, such a contribution could not be
sustained for longer than 4–5 days at most.
Although we did not conduct true energy balance
studies, a number of parameters indicated that a severely
catabolic state was induced in all groups that continued
to make significant quantities of milk. In particular, there
was a substantial loss of adipose tissue, as indicated by
the decrease in adipocyte size. Also, body weight was
unchanged in ad libitum fed animals ("2&3g; mean&
s.e.m.) but decreased by 42&3 g and 52&2 g in 50% or
25% ad libitum fed groups respectively (P<0·001 compared
with controls). In addition, serum concentrations of NEFA,
IGF-I, T3 and T 4 as well as the suppression of acetyl-CoA
carboxylase activity in adipose tissue and the degree of lipid
loss from adipose tissue were all consistent with increasing
catabolism. Acetyl-CoA carboxylase is considered to be the
rate-limiting step in lipogenesis and is acutely responsive to
levels of dietary intake (Volpe & Vagelos 1976). In contrast,
food restriction did not produce such a severely catabolic
state in rats that had their milk production inhibited by
treatment with anti-rGH and bromocriptine, since there
was no evidence of a major loss of lipid from adipose tissue
and serum T4 concentrations remained elevated. This
could be explained in part by the fact that rats treated with
anti-rGH and bromocriptine that were given ad libitum
access to food, despite reducing their food intake, con-
tinued to eat significantly more than non-lactating rats
(44&3 g (mean&s.e.m.) compared with non-lactating
values of approximately 20 g) even though they were only
producing very small quantities of milk. Even when food
intake was restricted to 25% ad libitum, i.e. to approxi-
mately 17 g compared with 68 g in lactating animals, this
only represents a little less than non-lactating animals
would normally eat.
GH plays an important role in milk production in both
ruminants (see Bauman & Vernon 1993) and rodents (Flint
et al. 1992), although it has been proposed that this eVect
is indirect, via IGF-I. As serum concentrations of IGF-I
generally reflect anabolic status, i.e. high on high planes of
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304 D J FLINT and R G VERNON · Food restriction and the milk yield response to prolactin and GH
nutrition and vice versa, IGF-I could serve as a monitor of
energy status modulating the response to exogenous GH
(McGuire et al. 1992, 1995). We examined this proposed
role for IGF-I in a rodent model in which the mammary
gland is sensitive to both GH and PRL. By reducing
food intake levels we hoped to produce an increasingly
catabolic state.
A graded reduction in food intake did lead to a graded
reduction in the response to GH in terms of both serum
IGF-I concentrations and milk yield. However, there was,
if anything, a greater eVect of food intake on the milk yield
response to PRL, despite unchanged serum IGF-I con-
centrations. In addition, the decrease in milk production
induced by restricting food intake in control lactating rats
was also achieved without any change in serum IGF-I
concentrations. Although it is possible that nutrient dep-
rivation is sensed in diVerent ways in determining the
response to GH and PRL, it seems more likely that they
use the same process and that this is not simply via IGF-I.
We cannot entirely rule out the possibility that IGF-I
plays some role in maintaining milk production since we
have recently shown that GH and PRL may interact
through the IGF system to maintain mammary cell survival
(Flint & Gardner 1994). Thus we have demonstrated that
PRL significantly inhibits the production of IGF-binding
protein-5 by the mammary gland, which we believe serves
to block IGF-I-mediated mammary cell survival and thus
allows apoptosis of the mammary epithelial cells to occur
(Tonner et al. 1995). It is interesting, however, to note in
the present study that, in animals rendered PRL- and
GH-deficient, there was a 30% loss of DNA over 48 h,
similar to our previous findings (Flint & Gardner 1994),
and that GH or PRL could prevent this completely inde-
pendently of nutritional intake. Further evidence for a
lack of influence of nutrition on mammary epithelial cell
survival was demonstrated in the control lactating animals,
since food deprivation, although dramatically decreasing
milk yield, did not influence mammary DNA content. It
thus seems that GH and PRL influence epithelial cell
survival and metabolic activity through diVerent pathways,
the former being largely independent of energy status with
the latter acutely responsive to it. Unravelling these diVer-
ent pathways is one of the major challenges in signal-
transduction science and this model may be of value in
achieving this goal.
If IGF-I does not serve as an indicator of energy balance
status, what does? A number of possibilities exist, including
insulin, T3, T4, blood glucose concentration (which is
known to influence milk yield because of the considerable
demand for glucose to synthesize milk lactose), NEFAs
(which increase on fasting and which are known to
suppress fatty acid synthesis in the mammary gland (Agius
& Williamson 1980)) and mammary blood flow (which is
also influenced by nutrition). Our data do not support a
role for any of these parameters as determinants of the
response of the mammary gland to GH and PRL since no
Journal of Endocrinology (1998) 156, 299–305
simple relationship was found between their concen-
trations in blood and milk yield response. This does not
exclude the possibility that local concentrations of, for
example, IGF-I and T3 could be important, since GH has
been shown to increase IGF-I mRNA in the mammary
gland (Kleinberg et al. 1990), and the mammary gland also
possesses a thyroxine 5*-deiodinase responsible for gener-
ating T3 within mammary tissue (Jack et al. 1994). Tissue
sensitivity to IGF-I could also be influenced through
nutrition by changes in IGF-I receptors which are present
on mammary epithelial cells (DehoV et al. 1988).
In summary we have shown that lactating rats continue
to produce large quantities of milk for at least 48 h despite
food restriction. This involves a number of adaptive
measures including reduced lipogenic potential and
marked lipid mobilization from adipose tissue and a
suppression of serum T4 concentrations. Serum insulin and
IGF-I concentrations are already decreased in lactation
and, probably as a consequence, these factors did not
decrease further upon feed restriction. The responses to
GH in terms of milk yield and the increase in serum IGF-I
showed a parallel decrease as feed intake was progressively
reduced, consistent with the hypothesis that the milk yield
response to GH is determined by the IGF-I response.
However, the fact that feed restriction also decreased
the milk yield response to PRL, and the milk yield of
untreated control rats, without any changes in serum
IGF-I concentrations, indicates that this hypothesis is
probably too simplistic, at least in the rat.
Acknowledgements
We thank M Gardner, E Finley, H Fawcett and E Tonner
for skilled technical assistance, and Mrs M Knight for
preparation of the manuscript. This study was funded
by the Scottish OYce Agriculture, Environment and
Fisheries Department.
References
Agius L & Williamson DH 1980 Rapid inhibition of lipogenesis in
vivo in lactating rat mammary gland by medium- or long-chain
triacylglycerols and partial reversal by insulin. Biochemical Journal 192
361–364.
Barber MC, Travers MT, Finley E, Flint DJ & Vernon RG 1992
Growth hormone–prolactin interactions in the regulation of
mammary and adipose tissue acetyl-CoA carboxylase activity and
gene expression in lactating rats. Biochemical Journal 285 469–475.
Bauman DE & Vernon RG 1993 EVects of exogenous bovine
somatotropin on lactation. Annual Review of Nutrition 13 437–461.
Bergmeyer HU, Bernt E, Schmidt F & Stork H 1974 Glucose:
determination with hexokinase and glucose 6-phosphate
dehydrogenase. In Methods of Enzymatic Analysis, vol 3, pp
1196–1201. Ed HU Bergmeyer. London: Academic Press Inc.
Borland CA, Barber MC, Travers MT & Vernon RG 1994 Growth
hormone inhibition of lipogenesis in sheep adipose tissue:
requirement for gene transcription. Journal of Endocrinology 142
235–242.
Downloaded from Bioscientifica.com at 03/19/2020 04:30:01PM
via free access
 Food restriction and the milk yield response to prolactin and GH
DehoV MH, Elgin RG, Collier RJ & Clemmons, DR 1988 Both
type I- and type-II insulin-like growth factor receptors increase
during lactogenesis in bovine mammary tissue. Endocrinology 122
33–39.
Flint DJ & Gardner MJ 1989 Inhibition of neonatal rat growth and
circulating concentrations of insulin-like growth factor-I using an
antiserum to rat growth hormone. Journal of Endocrinology 122
79–86.
Flint DJ & Gardner MJ 1994 Evidence that growth hormone
stimulates milk synthesis by direct action on the mammary gland
and that prolactin exerts eVects on milk secretion by maintenance
of mammary deoxyribonucleic acid content and tight junction
status. Endocrinology 135 1119–1124.
Flint DJ, Tonner E, Beattie J & Panton D 1992 Investigation of the
mechanism of action of growth hormone in stimulating lactation in
the rat. Journal of Endocrinology 134 377–383.
Grigor MR, Allan JE, Carrington JM, Carne A, Geursen A, Young
D, Thompson MP, Haynes EB & Coleman RA 1987 EVect of
dietary protein and food restriction on milk production and
composition, maternal tissues and enzymes in lactating rats. Journal
of Nutrition 117 1247–1258.
Itaya K & Ui M 1965 Colorimetric determination of free fatty acids in
biological fluids. Journal of Lipid Research 6 16–20.
Jack LJW, Kahl S, St Germain DL & Capuco AV 1994 Tissue
distribution and regulation of 5*-deiodinase processes in lactating
rats. Journal of Endocrinology 142 205–215.
Kanto U & Clawson AJ 1980 EVect of energy intake during
pregnancy and lactation on body composition in rats. Journal of
Nutrition 110 1829–1839.
Kleinberg DL, Ruan W, Catanese V, Newman CB & Feldman M
1990 Non-lactogenic eVects of growth hormone on growth and
insulin-like growth factor-1 messenger ribonucleic acid of rat
mammary gland. Endocrinology 126 3274–3276.
Labarca C & Paigen K 1980 A simple rapid and sensitive DNA assay
procedure. Analytical Biochemistry 102 344–352.
McGuire MA, Bauman DE, Miller MA & Hartnell GF 1992
Response of somatomedins (IGF-I and IGF-II) in lactating cows to
variations in dietary energy and protein and treatment with
recombinant n-methionyl bovine somatotropin. Journal of Nutrition
122 128–136.
· D J FLINT and R G VERNON 305
McGuire MA, Bauman DE, Dwyer DA & Cohick WS 1995
Nutritional modulation of the somatotropin/insulin-like growth
factor system: response to feed deprivation in lactating cows. Journal
of Nutrition 125 493–502.
Madon RJ, Ensor DM, Knight CH & Flint DJ 1986 EVects of an
antiserum to rat growth hormone on lactation in the rat. Journal of
Endocrinology 111 117–123.
Rasmussen KM 1992 The influence of maternal nutrition on lactation.
Annual Review of Nutrition 12 103–117.
Sainz RD, Calvert CC & Baldwin RL 1986 Relationships among
dietary protein, feed intake and changes in body and tissue
composition of lactating rats. Journal of Nutrition 116 1529–1539.
Sadurskis A, Sohlstrom A, Kabir N & Forsum E 1991 Energy
restriction and the partitioning of energy among the costs of
reproduction in rats in relation to growth of the progeny. Journal of
Nutrition 121 1798–1810.
Sampson DA & Jansen GR 1984 Measurement of milk yield in the
lactating rat from pup weight and weight gain. Journal of Pediatric
Gastroenterology and Nutrition 3 613–617.
Taylor JB, Calvert CC & Baldwin RL 1986 EVects of dietary protein,
fat and restriction on body composition and energy balance in
lactating rats. Journal of Nutrition 116 1519–1528.
Tonner E, Quarrie L, Travers M, Barber M, Logan A, Wilde C &
Flint D 1995 Does IGF-binding protein (IGFBP) present in the
involuting rat mammary gland regulate apoptosis? Progress in Growth
Factor Research 6 409–414.
Vernon RG 1980 Lipid metabolism in the adipose tissue of ruminant
animals. Progress in Lipid Research 19 23–106.
Vernon RG, Clegg RA & Flint DJ 1981 Metabolism of sheep adipose
tissue during pregnancy and lactation. Biochemical Journal 200
307–314.
Volpe JJ & Vagelos PR 1976 Mechanisms and regulation of
biosynthesis of saturated fatty acids. Physiological Reviews 56
339–417.
Received 13 February 1997
Revised manuscript received 6 June 1997
Accepted 1 September 1997
Journal of Endocrinology (1998) 156, 299–305
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