Biomedical subjects

M A Lomax

Publications and source records attributed to M A Lomax.

At least 19 recordsLinked to original sources

PubMed2005

The effect of dietary polyunsaturated fatty acids (PUFA) on infection with the nematodes Ostertagia ostertagi and Cooperia oncophora in calves.

Diet-induced changes in the polyunsaturated fatty acid (PUFA) content of immune cells influences the immune phenotype that develops following infection. The aim of this study was to examine the effect of manipulating dietary PUFA supply on tissue fatty acids composition and immunity to a mixed infection with an abomasal and an intestinal nematode parasite in calves. Calves (n=24) were allocated into two treatment groups and fed 25 g/day of either fish oil (n-3 group) or a binary mixture of palm/rapeseed oil (normal group) as a supplement in milk replacer. Within each treatment group eight calves were infected with 2000 L3 Ostertagia ostertagi and Cooperia oncophora, three times per week for 8 weeks, the remaining calves were pair-fed uninfected controls. Faecal egg counts (FEC) were carried out twice weekly. At slaughter, the whole gut was removed intact for worm counts and tissue samples were taken for fatty acid analysis. Samples of abomasum, duodenum and mid-gut were also collected for immunohistological analysis. FEC were not significantly influenced by oil supplement but tended to remain higher in the palm/rapeseed oil-fed group (normal infected). The number of intestinal immature worms was significantly (p<0.05) higher in the n-3 group. Mucosal mast cell (MMC) and eosinophil numbers were significantly increased (p<0.05) by infection and were significantly lower (p<0.05) in the intestinal tissue of the fish oil supplemented and infected group (n-3 infected group). These results suggest that feeding an n-3 PUFA-rich supplement (fish oil) can influence cellular mediators of immunity to nematode infection. This is the first report of the establishment of patency and the subsequent development of immunity to a mixed infection with O. ostertagi and C. oncophora in calves undergoing early rumen development. The trend in the FEC, MMC and eosinophil numbers in the n-3 group suggests that decreasing the dietary n-6/n-3 PUFA ratio may be a worthwhile immunonutritional strategy for potentiating the immune response to nematode parasite infection in the calf.

Animals↗

PubMed2005

Splanchnic metabolism of nitrogenous compounds and urinary nitrogen excretion in steers fed alfalfa under conditions of increased absorption of ammonia and L-arginine supply across the portal-drained viscera.

Effects of increased ammonia and/or arginine absorption across the portal-drained viscera (PDV) on net splanchnic (PDV and liver) metabolism of nitrogenous compounds and urinary N excretion were investigated in six catheterized Hereford x Angus steers (501 +/- 1 kg BW) fed a 75% alfalfa:25% (as-fed basis) corn-soybean meal diet (0.523 MJ of ME/[kg BW(0.75).d]) every 2 h without (27.0 g of N/kg of dietary DM) and with 20 g of urea/kg of dietary DM (35.7 g of N/kg of dietary DM) in a split-plot design. Net splanchnic flux measurements were obtained immediately before beginning and ending a 72-h mesenteric vein infusion of L-arginine (15 mmol/h). For 3 d before and during arginine infusion, daily urine voided was measured and analyzed for N composition. Feeding urea increased PDV absorption (P < 0.01) and hepatic removal (P < 0.01) of ammonia N, accounting for 80% of increased hepatic urea N output (P < 0.01). Numerical increases in net hepatic removal of AA N could account for the remaining portion of increased hepatic urea N output. Arginine infusion increased hepatic arginine removal (P < 0.01) and hepatic urea N output (P < 0.03) and switched hepatic ornithine flux from net uptake to net output (P < 0.01), but numerical changes in net hepatic removal of ammonia and AA N could not account fully for the increase in hepatic urea N output. Increases in urine N excretion equaled quantities of N fed as urea or infused as arginine. Estimated salivary urea N excretion was not changed by either treatment. Urea cycle regulation occurs via a complex interaction of mechanisms and requires N sources other than ammonia, but the effect of increased ammonia absorption on hepatic catabolism of individual AA in the present study was not significant.

Amino Acids↗

PubMed2005

Splanchnic metabolism of nutrients and hormones in steers fed alfalfa under conditions of increased absorption of ammonia and L-arginine supply across the portal-drained viscera.

Effects of increased ammonia and/or arginine absorption on net splanchnic (portal-drained viscera [PDV] plus liver) metabolism of nonnitrogenous nutrients and hormones in cattle were examined. Six Hereford x Angus steers (501 +/- 1 kg BW) prepared with vascular catheters for measurements of net flux across the splanchnic bed were fed a 75% alfalfa:25% (as-fed basis) corn and soybean meal diet (0.523 MJ of ME/[kg BW(0.75).d]) every 2 h without (27.0 g of N/kg of DM) and with 20 g of urea/kg of DM (35.7 g of N/kg of DM) in a split-plot design. Net flux measurements were made immediately before and after a 72-h mesenteric vein infusion of L-arginine (15 mmol/h). There were no treatment effects on PDV or hepatic O2 consumption. Dietary urea had no effect on splanchnic metabolism of glucose or L-lactate, but arginine infusion decreased net hepatic removal of L-lactate when urea was fed (P < 0.01). Net PDV appearance of n-butyrate was increased by arginine infusion (P < 0.07), and both dietary urea (P < 0.09) and arginine infusion (P < 0.05) increased net hepatic removal of n-butyrate. Dietary urea also increased total splanchnic acetate output (P < 0.06), tended to increase arterial glucagon concentration (P < 0.11), and decreased arterial ST concentration (P < 0.03). Arginine infusion increased arterial concentration (P < 0.07) and net PDV release (P < 0.10) and tended to increase hepatic removal (P < 0.11) of insulin, as well as arterial concentration (P < 0.01) and total splanchnic output (P < 0.01) of glucagon. Despite changes in splanchnic N metabolism, increased ammonia and arginine absorption had little measurable effect on splanchnic metabolism of glucose and other nonnitrogenous components of splanchnic energy metabolism.

3-Hydroxybutyric Acid↗

PubMed2004

An integrative model of amino acid metabolism in the liver of the lactating dairy cow.

The objective of this work was to construct a dynamic model of hepatic amino acid metabolism in the lactating dairy cow that could be parameterized using net flow data from in vivo experiments. The model considers 22 amino acids, ammonia, urea, and 13 energetic metabolites, and was parameterized using a steady-state balance model and two in vivo, net flow experiments conducted with mid-lactation dairy cows. Extracellular flows were derived directly from the observed data. An optimization routine was used to derive nine intracellular flows. The resulting dynamic model was found to be stable across a range of inputs suggesting that it can be perturbed and applied to other physiological states. Although nitrogen was generally in balance, leucine was in slight deficit compared to predicted needs for export protein synthesis, suggesting that an alternative source of leucine (e.g. peptides) was utilized. Simulations of varying glucagon concentrations indicated that an additional 5 mol/d of glucose could be synthesized at the reference substrate concentrations and blood flows. The increased glucose production was supported by increased removal from blood of lactate, glutamate, aspartate, alanine, asparagine, and glutamine. As glucose output increased, ketone body and acetate release increased while CO(2) release declined. The pattern of amino acids appearing in hepatic vein blood was affected by changes in amino acid concentration in portal vein blood, portal blood flow rate and glucagon concentration, with methionine and phenylalanine being the most affected of essential amino acids. Experimental evidence is insufficient to determine whether essential amino acids are affected by varying gluconeogenic demands.

Amino Acids↗

PubMed2004

Effect of plasma insulin and branched-chain amino acids on skeletal muscle protein synthesis in fasted lambs.

The increase in fractional rate of protein synthesis (Ks) in the skeletal muscle of growing rats during the transition from fasted to fed state has been explained by the synergistic action of a rise in plasma insulin and branched-chain amino acids (BCAA). Since growing lambs also exhibit an increase in Ks with level of feed intake, the objective of the present study was to determine if this synergistic relationship between insulin and BCAA also occurs in ruminant animals. Six 30 kg fasted (72 h) lambs (8 months of age) received each of four treatments, which were based on continuous infusion into the jugular vein for 6 h of: (1) saline (155 mmol NaCl/l); (2) a mixture of BCAA (0.778 micromol leucine, 0.640 micromol isoleucine and 0.693 micromol valine/min.kg); (3) 18.7 micromol glucose/min.kg (to induce endogenous insulin secretion); (4) co-infusion of BCAA and glucose. Within each period all animals received the same isotope of phenylalanine (Phe) as follows: (1) L-[1-13C]Phe; (2) L-phenyl-[ring 2H5]-alanine; (3) L-[15N]Phe; (4) L-[ring 2,6-3H]Phe. Blood was sampled serially during infusions to measure plasma concentrations of insulin, glucose and amino acids, and plasma free Phe isotopic activity; biopsies were taken 6 h after the beginning of infusions to determine Ks in m. longissimus dorsi and vastus muscle. Compared with control (saline-infused) lambs, Ks was increased by an average of 40% at the end of glucose infusion, but this effect was not statistically significant in either of the muscles sampled. BCAA infusion, alone or in combination with glucose, also had no significant effect on Ks compared with control sheep. Ks was approximately 60% greater for vastus muscle than for m. longissimus dorsi (P<0.01), regardless of treatment. It is concluded that there are signals other than insulin and BCAA that are responsible for the feed-induced increase in Ks in muscle of growing ruminant animals.

Amino Acids, Branched-Chain↗

PubMed2003

Consequences of exposure to serum, with or without vitamin E supplementation, in terms of the fatty acid content and viability of bovine blastocysts produced in vitro.

To determine whether serum supplementation influenced fatty acid content of bovine blastocysts and whether vitamin E addition to culture medium containing serum could improve development in vitro, cleaved eggs were cultured in synthetic oviduct fluid supplemented with bovine serum albumin (BSA, 0.4% w/v, fraction V) (SVBSA), fetal calf serum (FCS, 10% v/v) (SFCS) or FCS (10% v/v) plus 100 micro M vitamin E (SFCS + E). Blastocyst yields were recorded and fatty acid composition was determined by gas chromatography. Day 7 blastocysts were incubated with [2-(14)C] pyruvate for 3 h and then fixed for cell counts. Yields of good quality blastocysts were greatest from cleaved eggs cultured in serum-free conditions (P < 0.01). In the presence of serum, supplementation with vitamin E increased both total and good quality blastocyst yields (P < 0.01). Presence of serum increased fatty acid content (mean +/- SEM) of blastocysts (SVBSA v. SFCS = 57 +/- 2 v. 74 +/- 2 ng embryo(-1); P < 0.001). In contrast, pyruvate metabolism was greater in blastocysts produced without serum (27 +/- 3 v. 21 +/- 3 picomoles embryo(-1) 3h(-1); P < 0.01) but, on a per cell basis, no differences were detected. Addition of vitamin E to the serum-supplemented formulation did not alter either the fatty acid content (73 +/- 2 ng embryo(-1)) or pyruvate metabolism index (19 +/- 1 pmol embryo(-1) 3h(-1)) of SFCS + E blastocysts. Thus, despite lipid accumulation, supplementary vitamin E improved blastocyst yields in embryos exposed to serum.

Animals↗

PubMed2002

Nutrient-hormone interaction in the ovine liver: methionine supply selectively modulates growth hormone-induced IGF-I gene expression.

This study tested the hypothesis that specific amino acids are responsible for modulating the insulin-like growth factor-I (IGF-I) response to growth hormone (GH) in ovine hepatocytes. Cells were grown in media of defined amino acid composition, based on physiological concentrations (P.C.) of amino acids in sheep plasma. Relative to culture in 5 x P.C., amino acid limitation to 0.2 x P.C. had inhibitory effects on IGF-I RNA expression, peptide release and p70 S6 kinase phosphorylation (P<0.01 in each case). Limitation of methionine levels to 0.2 x P.C. against a background of 5 x P.C. for the other amino acids blocked GH-stimulated IGF-I peptide release and RNA expression, although basal expression was unaffected. In contrast, limitation of the other amino acids present in the culture medium had no effect on basal or GH-stimulated IGF-I expression. Selective methionine limitation to 0.2xP.C. levels had no effect on cellular or secretory protein synthesis rates relative to cells grown in complete 5 x P.C. medium but did cause a partial reduction in p70 S6 kinase phosphorylation, which was also observed when medium was selectively limited for other essential amino acids. The addition of rapamycin (5 ng/ml) to cells grown in 5xP.C. media completely abolished p70 S6 kinase phosphorylation (P<0.001), implicating mTOR in the response of S6 kinase phosphorylation to changing amino acid supply. By contrast, inclusion of rapamycin (100 ng/ml) had no effect on levels of IGF-I gene expression. These results indicate that methionine is the key limiting amino acid involved in the modulation of IGF-I expression in the ovine liver. This nutrient-hormone interaction is a highly selective phenomenon, occurring against a background of modest effects on general protein synthetic control. The partial inhibitory effects of methionine on mTOR activity are not sufficient to account for this selectivity of action.

Analysis of Variance↗

PubMed2000

Insulin stimulates phenylalanine uptake across the hind limb in fed lambs.

Five lambs ( approximately 6 mo of age and 30 kg), with an external iliac artery and vein catheterized and fed to maintain body weight, were used to examine effects of close arterial infusion of insulin and branched-chain amino acids (BCAA) on net phenylalanine (Phe) uptake across the hind limb. Treatments, administered randomly on five consecutive days to each lamb, were 400 min infusions of: i) saline (control); ii) insulin to double iliac artery concentration (low insulin); iii) as ii but to quadruple insulin concentration (high insulin); iv) 30 micromol/min leucine and 22.5 micromol/min each isoleucine and valine (BCAA) and v) co-infusion of ii and iv. Blood was sampled over the last 200 min from the iliac vein and right ventricle of the heart. High insulin caused a slight decrease (-13%, P < 0.05) in systemic glucose concentration, but did not alter systemic insulin concentration. Insulin, at both doses and in combination with BCAA, resulted in 9-fold greater net Phe uptake (P < 0.05) than the control, as did BCAA alone. Because BCAA alone increased net Phe uptake, these may have stimulatory effects directly or may enhance endogenous insulin. Maximum stimulation was achieved with low insulin because there was no increase in net Phe uptake with high insulin or from co-infusion with BCAA. Insulin, at low concentrations, may be important to growth in animals with marginal nutrition.

Amino Acids, Branched-Chain↗

PubMed2000

Effect of feeding level and thyroxine on adipose tissue development and growth in postnatal lambs.

We investigated the influence of exogenous thyroxine (T4) administration in conjunction with level of feeding on adipose tissue and liver growth in postnatal lambs. Pairs of lambs were fed either 100 g (i.e. low fed) or 200 g (i.e. high fed) of milk powder per litre of reconstituted milk replacer over the first month of life. Half of the pairs of lambs were fed a bolus dose of T4 (15 mg (kg body weight)-1) daily until 8 days of age. Perirenal adipose tissue and hepatic tissue were sampled at either 8 or 35 days of age. High fed lambs grew faster, possessed more adipose tissue and had larger livers than low fed lambs at 8 and 35 days of age. T4 administration resulted in a lower thermogenic activity (i.e. GDP binding) in adipose tissue at 8 days of age in low, but not high fed lambs. There was no difference between groups in colonic temperature or oxygen consumption. Between 17 and 35 days of age high fed lambs previously treated with T4 had lower daily milk consumption than untreated siblings, but still attained the same growth rate. Plasma insulin-like growth factor-I concentrations were greater in high than low fed lambs, a relationship that was not influenced by T4 treatment. In adipose tissue, iodothyronine 5' deiodinase activity was not influenced by T4 administration and was greater in high than low fed lambs. Hepatic iodothyronine 5' deiodinase activity was not influenced by T4 administration in low fed lambs, but was reduced by T4 in high fed lambs. In conclusion, T4 administration over the first 8 days of life can accelerate the rate of decline in thermogenic activity of uncoupling protein-1. This effect is not observed when the level of feeding is increased. Following withdrawal of T4 treatment, high fed T4-treated lambs were able to maintain the same growth rate as untreated lambs despite having a lower food intake.

Adipose Tissue↗

PubMed1999

Chronic cold exposure has no effect on brown adipose tissue in newborn lambs born to well-fed ewes.

It has been previously shown in twin-bearing ewes fed only 60% of their metabolizable energy requirements for late pregnancy that chronic cold exposure induced by winter shearing of the ewes results in larger lambs with more brown adipose tissue. This effect appears to be primarily due to prevention of a decline in fetal body and tissue weights between 145 days' gestation and 2 h after birth (i.e. 147 days' gestation) in lambs born to underfed shorn ewes. The present study therefore examined the impact, in ewes that were well fed (i.e. received 100% of their metabolizable energy requirements) during the final month of gestation, of chronic cold exposure induced by winter shearing on lamb birthweight and perirenal adipose tissue composition as measured 2 h after birth. Perirenal adipose tissue was analysed for its thermogenic activity (i.e. GDP binding to mitochondria) and catecholamine content. These observations were combined with similar measurements made in near-term (i.e. 145 days' gestation) fetuses sampled from well-fed unshorn ewes. There was no difference between lambs born to shorn or unshorn ewes with respect to birthweight or perirenal adipose tissue weight and composition. Perirenal adipose tissue weight was higher in lambs born to unshorn ewes than in fetuses. The thermogenic activity of adipose tissue was 2-fold higher in lambs born to unshorn ewes compared with 145-day-old fetuses. Epinephrine was detectable only at very low levels in fetal perirenal adipose tissue, increasing 10-fold after birth, with no difference between lambs born to shorn or unshorn ewes. In newborn lambs, plasma growth hormone concentration was lower and insulin concentration higher in shorn compared with unshorn groups. In conclusion, chronic cold exposure induced by winter shearing had no effect on brown adipose tissue development in lambs born to well-fed ewes.

Adipose Tissue, Brown↗

PubMed1999

Growth hormone and amino acid supply interact synergistically to control insulin-like growth factor-I production and gene expression in cultured ovine hepatocytes.

Many of the anabolic effects of growth hormone (GH) are indirect, occurring through GH-stimulated production of insulin-like growth factor-I (IGF-I) by the liver. As well as being regulated by GH, plasma IGF-I concentrations have been demonstrated to depend upon the level of dietary protein intake, with low protein diets being associated with reduced circulatory IGF-I levels. This inhibitory effect cannot be reversed by GH injection, suggesting that liver sensitivity to GH becomes impaired.To investigate the mechanisms through which protein supply affects GH sensitivity, primary cultures of ovine hepatocytes were grown in defined media, containing various proportions (0.2, 1.0 and 5.0) of jugular amino acid concentrations in fed sheep. Production of IGF-I by these cells was measured after 24 and 48 h in culture by radioimmunoassay. In the first 24-h period basal IGF-I production was the same in all defined media, and GH caused an approximately 2-fold increase in IGF-I release in cells grown in 1.0xor 5.0xamino acid media (P<0.01). Although GH appeared to increase IGF-I release in this period for cells grown in 0.2xamino acid media, this effect was not statistically significant. In the period from 24-48 h in defined media, both basal and GH-stimulated IGF-I production was dependent on amino acid availability (P<0.05 and P<0.001 respectively). Factorial analysis of variance demonstrated a strong positive interaction (P<0.001) between the effects of amino acid availability and GH, such that GH increased IGF-I production by more than 2-fold in cells grown in 5.0xamino acid media (P<0.01) but had no effect on production by cells grown in 1.0xor 0.2xamino acid media. Measurement of steady state concentrations of exon 1-derived IGF-I mRNAs using an RNase protection assay demonstrated that the observed effects on IGF-I peptide secretion were strongly associated with parallel effects at the RNA level. Incorporation of (35)S-methionine into cellular proteins over a 4-h period starting 20 h after transfer to defined culture media was not significantly reduced in 1.0xcompared with 5.0x amino acid media, although rates under both of these conditions were significantly higher than those seen in 0.2xamino acid media (P<0.01). The lack of correspondence between the dose-dependent effects of amino acid supply on cellular protein synthesis and those on basal and GH-stimulated IGF-I production, suggests that amino acid supply modulates IGF-I production through selective mechanisms. Steady state levels of the CCAAT/enhancer-binding protein beta (C/EBPbeta) isoforms, liver-enriched activating protein (LAP) and liver-enriched inhibitory protein (LIP) were determined by Western blotting. When levels of LAP were expressed relative to LIP levels in the same extracts, a significant decrease in the LAP:LIP ratio was observed in response to amino acid limitation (P<0.05). These data strengthen earlier arguments that synergistic interaction between the effects of amino acids and GH on hepatic IGF-I gene expression underlie nutrition-dependent changes in circulating IGF-I titres. The association between these effects and altered levels of C/EBPbeta isoforms suggests that CCAAT/enhancer mediated control of IGF-I gene expression may be involved in this phenomenon.

Amino Acids↗

PubMed1998

An immunohistochemical and in situ hybridisation study of the postnatal development of uncoupling protein-1 and uncoupling protein-1 mRNA in lamb perirenal adipose tissue.

In the lamb, the uncoupling protein-1 (UCP1) content of perirenal adipose tissue at birth is an important factor in heat production by non-shivering thermogenesis and the prevention of hypothermia. This study examines UCP1 gene expression and protein content in perirenal adipose tissue over the first 15 days of life by in situ hybridisation and immunohistochemistry. UCP1 mRNA was detected at birth in 30% of adipocytes, and in approximately 24% of fat cells at 2 days of life. However, by 5 days of age and thereafter UCP1 mRNA was undetectable. Immunoreactive UCP1 was present in all adipocytes at birth and at 2 days of age, and remained detectable in a decreasing proportion of cells until day 10 of life. By 15 days of age no immunoreactive UCP1 was detected and the perirenal adipose tissue had the appearance of white fat. It is concluded that UCP1 gene expression is suppressed in most adipocytes in perirenal adipose tissue of newborn lambs, and gene expression rapidly falls in the remaining adipocytes over the first 5 days of postnatal life. In contrast, immunoreactive UCP1, a characteristic of brown adipose tissue, was present in many adipocytes for up to 10 days of age, suggesting that UCP1 has a long half-life in lambs. All adipocytes in perirenal adipose tissue of newborn lambs appear to be functionally brown, but over the first 2 weeks of postnatal life there is a complete transformation to white adipocytes.

Adipose Tissue↗

PubMed1997

Substrate specificity of the mammary tissue anionic amino acid carrier operating in the cotransport and exchange modes.

The substrate specificity of the rat mammary tissue high affinity, Na+-dependent anionic amino acid transport system has been investigated using explants and the perfused mammary gland. D-Aspartate appears to be transported via the high affinity, Na+-dependent L-glutamate carrier. Thus, D-aspartate transport by rat mammary tissue was Na+-dependent and saturable with respect to extracellular D-aspartate with a Km and Vmax of 32.4 microM and 49.0 nmol/2 min per g of cells respectively. The uptake of D-aspartate by mammary explants was cis-inhibited by L-glutamate and L-aspartate, but not by D-glutamate. L-glutamate uptake by mammary tissue explants was cis-inhibited by beta-glutamate, L-cysteate, L-cysteine sulfinate and dihydrokainate but not by DL-alpha-aminoadipate. In addition, dihydrokainate, but not DL-alpha-aminoadipate inhibited D-aspartate and L-glutamate uptake by the perfused gland. D-Aspartate efflux from mammary tissue explants was trans-accelerated by external L-glutamate in a dose-dependent fashion (50-500 microM). The effect of L-glutamate on D-aspartate efflux was dependent on the presence of extracellular Na+. D-Aspartate, L-aspartate and L-cysteine sulfinate (at 500 microM) also markedly trans-stimulated D-aspartate efflux from mammary tissue explants. In contrast, L-cysteine. D-glutamate, L-leucine, dihydrokainate and DL-alpha-aminoadipate were either weak stimulators of D-aspartate efflux or were without effect. D-Aspartate efflux from the perfused mammary gland was trans-stimulated by L-glutamate but not by D-glutamate and only weakly by L-cysteine (all at 500 microM). It appears that the mammary tissue high affinity anionic amino acid carrier can operate in the exchange mode with a similar substrate specificity to that of the co-transport mode.

Amino Acid Transport System X-AG↗

PubMed1997

Mammary protein synthesis is acutely regulated by the cellular hydration state.

The effect of cell-volume pertubations on mammary tissue protein synthesis has been examined. Cell-swelling, induced by a hyposmotic shock, increased the rate of incorporation of radiolabelled leucine and methionine into trichloroacetic acid precipitable material. The incorporation of radiolabel under both isosmotic and hyposmotic conditions was inhibited by cycloheximide. The increases in mammary protein synthesis as a result of cell-swelling may be attributable to an increase in casein synthesis. Conversely, cell-shrinking, as a consequence of a hyperosmotic challenge, almost abolished mammary protein (casein) synthesis. The finding that cell-volume pertubations had no significant effect on steady-state casein mRNA levels suggests that the regulation, within the time course of the experiments, is at the level of translation. The results strongly suggest that mammary cell volume may be an important cellular signal in the control of mammary protein synthesis in general and casein synthesis in particular.

Animals↗

PubMed1997

Maternal manipulation of brown adipose tissue and liver development in the ovine fetus during late gestation.

We examined the effect of maternal chronic cold exposure, induced by winter-shearing ewes 4 weeks before their predicted lambing date, on brown adipose tissue (BAT) and liver development in lambs. Fetuses were sampled from under-fed (60% of energy requirements for maintenance and pregnancy of an unshorn ewe) shorn or unshorn ewes at 126, 140 and 145 d of gestation. Lambs were sampled from ewes within 2 h of birth. Throughout gestation fetal body, BAT and liver weights were similar in shorn and unshorn groups. The level of GDP binding to mitochondrial uncoupling protein remained low throughout gestation, but increased dramatically after birth. Lambs born to shorn ewes possessed more mitochondrial protein and exhibited a significantly higher total thermogenic activity in BAT. Type I iodothyronine 5' deiodinase (EC 3.8.1.4) activity in BAT peaked at birth, as did hepatic iodothyronine 5' deiodinase activity and was significantly greater in lambs born to under-fed shorn ewes, which exhibited a higher plasma triiodothyronine concentration. Chronic maternal adaptations to prolonged cold exposure appear to enable pregnant ewes to compensate for the negative effects of under-feeding on fetal growth and development.

Adaptation, Physiological↗

PubMed1997

Adipose tissue development during early postnatal life in ewe-reared lambs.

This study examines the precise time course that brown adipose tissue (BAT) takes to adopt the characteristics of white adipose tissue in postnatal lambs. Perirenal adipose tissue was sampled from ewe-reared lambs within 1 h of birth and at 1, 2, 4, 7, 14, 21 and 30 days of age and analysed for the amount of mRNA for uncoupling protein (UCP), the amount and activity of UCP, and protein, mitochondrial protein and lipid content. This was combined with measurements of colonic temperature and jugular venous plasma concentrations of thyroid hormones and insulin-like growth factor-1 (IGF-1). Over the first 4-7 days of age, large quantities of UCP mRNA were associated with a peak in plasma triiodothyronine concentration at 2 days of age followed by a maximal amount and activity of UCP at 4 days and a basal colonic temperature of 39.3 degrees C. Between 7 and 30 days there was a large increase in lipid deposition as the amount and activity of UCP and the amount of UCP mRNA declined to basal values and colonic temperature was maintained at 40 degrees C. A significant positive relationship between perirenal adipose tissue lipid content and plasma IGF-1 concentration was observed throughout the study period. It is concluded that ovine adipose tissue maturation occurs in two distinct phases over the first month of life. The precise time scale of this process could be regulated in part by the lamb's body temperature which determines whether adipose tissue is required for heat production (i.e. BAT) or as an endogenous energy source (i.e. white adipose tissue).

Adipose Tissue↗

PubMed1996

The mechanism of L-glutamate transport by lactating rat mammary tissue.

The transport of L-glutamate by lactating rat mammary gland has been examined using both tissue explants and a perfused mammary preparation. L-Glutamate uptake by mammary tissue explants was predominantly via a Na(+)-dependent pathway: Li+, choline+ and NMDG+ could not substitute for Na+. L-Glutamate efflux from preloaded explants was also influenced by the transmembrane Na(+)-gradient. These results are consistent with (Na(+)-glutamate) cotransport. The Na(+)-dependent system for L-glutamate transport in tissue explants was saturable (Km = 112.5 +/- 19.7 microM; Vmax = 71.3 +/- 10.4 nmol/min per g cells) and selective for anionic amino acids. Thus, D- and L-aspartate were high affinity inhibitors of L-glutamate uptake whereas neutral amino acids were relatively ineffective. D-Aspartate inhibited L-glutamate uptake in a competitive fashion. L-Glutamate uptake by the perfused mammary gland was (a) Na(+)-dependent (b) saturable (Km = 18.1 +/- 4.9 microM; Vmax = 40.3 +/- 3.7 nmol/min per g tissue) and (c) selective for anionic amino acids. The results suggest that the (Na(+)-glutamate) cotransporter is situated in the blood-facing aspect of the mammary epithelium.

Amino Acid Transport System X-AG↗

PubMed1996

Leucine and protein metabolism in the lactating dairy cow mammary gland: responses to supplemental dietary crude protein intake.

Mammary gland protein metabolism, determined by an arteriovenous difference technique, was monitored in four Holstein-Friesian dairy cows in response to supplemental dietary protein (provided as rumen-protected soyabean meal) during late lactation (weeks 24-30). Each cow was offered two isoenergetic diets composed of grass silage (170 g crude protein/kg dry matter) plus either a low (108 g/kg) or medium (151 g/kg) crude protein concentrate in a single crossover design involving two 21 d periods. On day 21, arteriovenous measurements across the mammary gland were made during a 13 h continuous i.v. infusion of [1-(13)C]leucine and with frequent (2 hourly) milk sampling during the final 6 h. Although total milk yield was slightly increased (+1 kg/d) by protein supplementation, milk protein yield was not significantly affected. Whole body protein flux (protein synthesis plus oxidation) was not significantly affected by supplementation. Total mammary gland protein synthesis (milk plus non-milk protein) was also not affected by supplementation but on both diets gland synthesis was always greater (by 20-59 percent) than milk protein output. The fractional oxidation rate of leucine by the mammary gland was significantly increased by protein supplementation (0-047 v. 0-136). Although the enrichment of leucine in secreted milk protein continued to increase, the final value (at 13 h) was 0-94 of the arterial plasma free leucine plateau value (not significantly different), suggesting almost exclusive use of plasma free leucine for milk protein synthesis. Based on current feeding schemes for dairy cattle, a fixed proportion (0-65 0-75) of the additional protein intake (+490 g/d) should have been partitioned into milk protein. Instead, leucine oxidation by the mammary gland was increased. Whether oxidation of other amino acids was also enhanced is unknown but if amino acid oxidation and the 'additional' non-milk protein synthesis occurring in the gland are not crucial to milk synthesis, then by reducing such activities improvements in the efficiency of converting absorbed amino acid into milk protein can be achieved.

Animals↗