Biomedical subjects

S S Donkin

Publications and source records attributed to S S Donkin.

At least 19 recordsLinked to original sources

PubMed2006

Effects of short-term glucagon administration on gluconeogenic enzymes in the liver of midlactation dairy cows.

During lactation, the dairy cow experiences an increased demand for glucose to support milk production. Increased glucose demand can be met through increased capacity for gluconeogenesis, increased supply of glucose precursors, or a combination of both processes. Glucagon, a key hormone in glucose homeostasis, acts to promote gluconeogenesis and increase glucose output from liver. The objective of this study was to determine the effect of short-term administration of glucagon on expression of gluconeogenic enzymes in lactating dairy cattle. Sixteen multiparous Holstein cows were selected from the Purdue University Animal Sciences Dairy Research Center herd. Cows were stratified on the basis of milk production and days in milk and randomly assigned to either a saline or glucagon injection group (n = 8 per group). Cows were injected subcutaneously at -21, -14, -7, and 0 h relative to final glucagon and saline injections with either 3.75 mg of lyophilized bovine glucagon (15 mg/d) dissolved in 60 mL of 0.15 M NaCl (pH 10.25) or 60 mL of 0.15 M NaCl. Liver biopsy samples were obtained 1 wk before injection to establish baseline values and at 3 h after cows received final glucagon and saline injections. Biopsy samples were analyzed for mRNA abundance, enzyme activity, protein abundance, and in vitro measures of gluconeogenesis. Glucagon did not alter pyruvate carboxylase or cytosolic phosphoenolpyruvate carboxykinase (PEPCK) mRNA abundance, enzyme activity, or protein abundance, although there was a tendency for greater mRNA expression with the glucagon treatment (4.69 vs. 6.78, arbitrary units). Glucagon injections did not change mitochondrial PEPCK mRNA expression. Gluconeogenesis from 2.5 mM [2-(14)C]propionate and 2.0 mM [U-(14)C]lactate was similar in liver biopsy samples from glucagon-treated and control cows. There was no effect of glucagon on dry matter intake and milk production. Glucose, nonesterified fatty acids, beta-hydroxybutyrate acid, and insulin were not altered by glucagon. Blood glucagon was elevated, 76.09 vs. 96.14 pg/mL, for cows receiving glucagon injections. The data indicate that 24-h administration of glucagon does not alter cytosolic PEPCK mRNA expression or result in immediate alterations in total PEPCK enzyme activity and gluconeogenic capacity.

3-Hydroxybutyric Acid↗

PubMed2006

Major advances in fundamental dairy cattle nutrition.

Fundamental nutrition seeks to describe the complex biochemical reactions involved in assimilation and processing of nutrients by various tissues and organs, and to quantify nutrient movement (flux) through those processes. Over the last 25 yr, considerable progress has been made in increasing our understanding of metabolism in dairy cattle. Major advances have been made at all levels of biological organization, including the whole animal, organ systems, tissues, cells, and molecules. At the whole-animal level, progress has been made in delineating metabolism during late pregnancy and the transition to lactation, as well as in whole-body use of energy-yielding substrates and amino acids for growth in young calves. An explosion of research using multicatheterization techniques has led to better quantitative descriptions of nutrient use by tissues of the portal-drained viscera (digestive tract, pancreas, and associated adipose tissues) and liver. Isolated tissue preparations have provided important information on the interrelationships among glucose, fatty acid, and amino acid metabolism in liver, adipose tissue, and mammary gland, as well as the regulation of these pathways during different physiological states. Finally, the last 25 yr has witnessed the birth of "molecular biology" approaches to understanding fundamental nutrition. Although measurements of mRNA abundance for proteins of interest already have provided new insights into regulation of metabolism, the next 25 yr will likely see remarkable advances as these techniques continue to be applied to problems of dairy cattle biology. Integration of the "omics" technologies (functional genomics, proteomics, and metabolomics) with measurements of tissue metabolism obtained by other methods is a particularly exciting prospect for the future. The result should be improved animal health and well being, more efficient dairy production, and better models to predict nutritional requirements and provide rations to meet those requirements.

Absorption↗

PubMed2005

Effects of dexamethasone and growth hormone treatment on hepatic gluconeogenic enzymes in calves.

The hypothesis was tested that dexamethasone (DX) and bovine somatotropin (bST) alter expression or activity of gluconeogenic enzymes in neonatal calves. Holstein dairy calves (n = 24) were randomly divided in 4 groups and were treated with saline (control group), with DX at 30 microg/kg body weight per d (CDX), with 500 mg of sustained-release recombinant bST every 14 d (CbST), and with the combination of DX and bST from d 3 through 42 of life (CbSTDX). Plasma glucose and insulin concentrations were elevated throughout the study in CbSTDX, and insulin concentrations were elevated in CDX from d 7 to 28. Treatment with DX and the combination of DX and bST increased plasma glucagon concentrations from d 14 to 42, but decreased plasma cortisol concentrations on d 7 and 14 when compared with control calves. In liver, phosphoenolpyruvate carboxykinase (PEPCK) mRNA levels were reduced in CDX and CbSTDX when compared with control calves or CbST. The activity of PEPCK on d 14 was higher in CbSTDX compared with control calves. Pyruvate carboxylase mRNA levels were decreased on d 7 in CDX and CbSTDX. Pyruvate carboxylase activities on d 14 and 28 were lower in CDX and CbSTDX than in control calves or CbST. These data indicate an age-dependent response to DX for blood metabolites, expression and activities of hepatic PEPCK and pyruvate carboxylase, and for effects of bST, suggesting that glucocorticoid status is important.

Animals↗

PubMed2005

Feed restriction induces pyruvate carboxylase but not phosphoenolpyruvate carboxykinase in dairy cows.

The ability of dairy cattle to adapt to changes in nutrient intake requires appropriately responsive expression of several key genes in liver. Holstein cows were used in 2 experiments to determine the effect of short-term feed restriction on expression of mRNA for gluconeogenic and ureagenic enzymes in liver. In experiment 1, cows were fed a total mixed diet for ad libitum intake for a 5-d period followed by 5 d of 50% of their previous 5-d ad libitum intake followed by 10 d of ad libitum feeding. Liver biopsies and blood samples were obtained on d 5, 10, and 20 of the experiment, the last day of each feeding period. Pyruvate carboxylase (PC) mRNA increased with feed restriction, but phosphoenolpyruvate carboxykinase (PEPCK) was unchanged. Expression of carbamoyl phosphate synthetase (CPS-I), argininosuccinate synthetase, and ornithine transcarbamylase mRNA were not altered by feed restriction; however, CPS-I mRNA expression tended to increase during realimentation. In experiment 2, cows were fed for ad libitum intake for 5 d and then fed 50% of previous intake for 5 d. Liver biopsy samples collected on d 5 and 10 were used for PC mRNA, PEPCK mRNA, and in vitro measure of gluconeogenesis from radiolabelled propionate and lactate. The data indicate expression of genes for key metabolic processes in liver of lactating cows is responsive to feeding level. Expression of PC mRNA is part of the adaptive response to feed intake restriction and is matched by increased capacity for gluconeogenesis from lactate.

Animals↗

PubMed2004

Toll-like receptors 2 and 4, and acute phase cytokine gene expression in dexamethasone and growth hormone treated dairy calves.

Cattle are exposed to growth hormone stimulants and to stressors that cause cortisol release. Both of these hormones affect immune responses which may reduce disease resistance. Toll-like receptors are the pattern recognition molecules of pathogens that are on immune cells. They then orchestrate the induction of the appropriate acute phase cytokines of the early innate response. The objective of this study was to determine changes in toll-like receptors and acute phase cytokines following treatment with a synthetic glucocorticoid (dexamethasone) and growth hormone (GH). Twenty-eight calves were given the control (Cnt), dexamethasone (DEX), GH, or dexamethasone and GH (Both) treatments from 3 until 56 days of age. Blood was collected by jugular venipuncture on days 14, 28, 42, and 56. On day 56, a lung lavage was performed and spleen and thymus tissues collected. Total RNA was extracted from blood leukocytes, lung lavage cells, spleen and thymus cells. Real-time reverse transcriptase-polymerase chain reaction (RT-PCR) was used to quantify interleukin-1 (IL-1), IL-1 receptor antagonist (IL-1Ra), tumor necrosis factor (TNF)-alpha, toll-like receptor 2 (TLR2), and toll-like receptor 4 (TLR4). Blood leukocytes had a time effect for IL-1Ra (P < 0.01), with a trend for a treatment effect (P = 0.07) and had a treatment by time interaction (P < 0.05). IL-1, TNF, and TLR2 and TLR4 were greatest (P < 0.05) for Cnt only at day 14. IL-1 expression of lung lavage cells was greatest (P < 0.05) for calves on the Both treatment compared to the other three treatments. However, IL-1Ra was not different among the treatments. Toll-like receptor 2 expression was enhanced with Both compared to either DEX (P < 0.05) or GH (P < 0.05) and tended to be greater than Cnt expression (P = 0.07). Expression of TLR4 tended to be reduced by Both compared to Cnt (P = 0.06). Tumor necrosis factor-alpha was greatly enhanced by Both compared to the other three treatments (P < 0.05). Spleen cell tended to have different IL-1 expression between GH and Both (P < 0.10). Interleukin-1 receptor antagonist and TLR2 and TLR4 were not different among treatments. However, TNF-alpha expression was enhanced by the DEX treatment alone compared to the GH treatment (P < 0.05), and tended (P < 0.10) to be greater than Cnt expression. None of the gene expressions were different among treatments for thymus cells. Lung lavage cell expression appears to be most susceptible to these hormones while blood leukocyte expression was only slightly affected, and thymus cells were not affected at all. These data demonstrate that TLR2 and TLR4 and acute phase cytokine expression can be altered by stress and growth hormones, which may decrease resistance of those animals to disease.

Animals↗

PubMed2004

Bovine somatotropin increases hepatic phosphoenolpyruvate carboxykinase mRNA in lactating dairy cows.

Somatotropin (ST) increases milk production and through coordinated changes in hepatic glucose synthesis and amino acid metabolism in dairy cows. The objective of this study was to determine the effects of ST on hepatic mRNA expression for phosphoenolpyruvate carboxykinase (PEPCK) and pyruvate carboxylase (PC), enzymes that are critical to the synthesis of glucose in liver and hepatic mRNA expression for carbamylphosphate synthetase I (CPS-I), argininosuccinate synthetase (AS), and ornithine transcarbamylase (OTC), critical enzymes of the urea cycle. Eighteen cows were randomly allocated to 2 treatment groups and received either recombinant bovine ST (Posilac; Monsanto, St. Louis, MO) or saline injections at 14-d intervals during a 42-d period. Expression of mRNA was determined using Northern blot analysis. Nuclei, isolated from liver biopsy samples, were used to determine effects of ST on transcription rate of PEPCK. Milk production was increased with ST (37.3 vs. 35.1+/-0.6 kg/ d). Plasma NEFA was increased with ST (299 vs. 156+/-34 microM). There were no differences in the expression of CPS-I, AS, and OTC mRNA with ST. Expression of PEPCK and IGF-I mRNA were increased with ST but PC mRNA was unchanged. The data indicate increased PEPCK mRNA in cows given ST and indicates a greater capacity for gluconeogenesis from gluconeogenic precursors that form oxaloacetate. The effects of ST to elevate PEPCK mRNA expression require chronic administration and involve increased transcription of the PEPCK gene.

Animals↗

PubMed2003

The response of the hepatic insulin-like growth factor system to growth hormone and dexamethasone in calves.

Glucocorticoids inhibit postnatal growth and yet can stimulate the somatotropic axis around birth. The aim of the present study was to investigate the effects of dexamethasone on the somatotropic axis and on the responses of the insulin-like growth factor (IGF) system to growth hormone treatment in calves. Calves (n=24) were randomly divided into four groups. Group DX was injected with dexamethasone (30 micro g/kg body weight per day), group GH was injected with 500 mg slow-release bovine growth hormone at 14-day intervals, group GHDX was injected with dexamethasone and bovine growth hormone, and group CNTRL (serving as control) was injected with saline from day 3 to day 42 of life. Blood samples were taken on day 3 and blood and liver samples were obtained on days 7, 14, 28 and 42. Body weight increased in the CNTRL and GH groups up to the end of the study and in the DX and GHDX groups up to the fourth week. Dexamethasone treatment decreased (P<0.05) plasma IGF binding protein (IGFBP)-1 on days 7 and 14, but increased (P<0.05) plasma IGFBP-1, decreased (P<0.05) plasma IGF-I and IGFBP-3, and decreased hepatic mRNA for growth hormone receptor (GHR) and IGF-I on day 42. Growth hormone treatment increased (P<0.05) plasma growth hormone concentrations on days 7 and 14, tended to increase (P<0.1) plasma IGF-I concentrations on day 42, and increased (P<0.05) hepatic mRNA levels of GHR on day 14 and IGF-I mRNA levels on days 7 and 14. The combined dexamethasone and growth hormone treatment increased plasma growth hormone concentrations on day 7 and resulted in the highest plasma concentrations of IGF-I and IGFBP-3 (day 7 to day 28) as well as the greatest abundance of hepatic GHR (day 14) and IGF-I (days 7 and 14) mRNA. Plasma IGFBP-1 concentrations in the GHDX group behaved in a similar manner as in the DX group. In conclusion, the response of the somatotropic axis to growth hormone treatment could be greatly enhanced by dexamethasone treatment during the neonatal and early postnatal period, but body weight gain was not improved. Dexamethasone alone inhibited the somatotropic axis and postnatal growth after the first Month of life.

Animals↗

PubMed2003

Effects of feeding silage and grain from glyphosate-tolerant or insect-protected corn hybrids on feed intake, ruminal digestion, and milk production in dairy cattle.

Lactating dairy cows were used to determine effects of feeding glyphosate-tolerant or insect-protected corn hybrids on feed intake, milk production, milk composition, and ruminal digestibility. Corn resistant to European corn borer (Ostrinia nubilalis) infestation (Bt-MON810), or its nontransgenic control (Bt-CON), were planted in alternating fields during two successive years. One-half of each strip was harvested for whole plant corn silage and the remainder was allowed to mature and harvested as grain. Effects of feeding diets containing either Bt-MON810 or Bt-CON grain and silage were determined in two experiments (1 and 2) conducted during successive years. In experiment 3, glyphosate-tolerant Roundup Ready corn (RR-GA21) or its nontransgenic control (RR-CON) corn were grown in alternating fields during one cropping season. Diets contained 42 to 60% corn silage and 20 to 34% corn grain from Bt-MON810, RR-GA21, or the appropriate nontransgenic counterpart; treatments were applied using a switchback design. Cows were fed ad libitum and milked twice daily. There were no differences for nutrient composition between silage sources or between grain sources within an experiment. Data for experiments 1 and 2 indicated similar dry matter intake (DMI), 4% fat-corrected milk (FCM) production, and milk composition between Bt-MON810 and Bt-CON diets. There were no differences for DMI, 4% FCM production, and milk composition between RR-GA21 and RR-CON diets. There was no difference in ruminal degradability, determined separately for corn silage and corn grain, for RR-GA21 or Bt-MON810-hybrids compared with their respective controls. These data demonstrate equivalence of nutritional value and production efficiency for corn containing Bt-MON810 compared with its control and for RR-GA21 corn compared with its control.

Animal Feed↗

PubMed2001

Rumen undegradable protein, rumen-protected choline and mRNA expression for enzymes in gluconeogenesis and ureagenesis in periparturient dairy cows.

The objective of this study was to profile mRNA expression of argininosuccinate synthetase (AS) and ornithine transcarbamylase (OTC), two enzymes that participate in the formation of urea in liver and compare these with changes in mRNA for pyruvate carboxylase (PC) and phosphoenolpyruvate carboxykinase (PEPCK) during the periparturient period in dairy cows. Forty-eight multiparous Holstein cows were fed isoenergetic prepartum diets that contained 10% RDP and either 4.0% RUP or 6.2% RUP and either 0, 6, or 12 g/d of rumen-protected choline (RPC) as CapShure (Balchem Corp., Slate Hill, NY). After calving cows received a common diet and continued RPC as per their prepartum assignments. Liver biopsies were obtained on d -28, -14, 1, 28, and 56 relative to calving, and the abundances of AS, OTC, PC, PEPCK, and 18S mRNA were determined by Northern blot analysis of total RNA. The abundance of OTC mRNA was lowest at calving and was decreased by RPC and 6.2% RUP feeding. Feeding 6.2% RUP did not alter AS, PC, or PEPCK mRNA. The expression of AS mRNA increased and PEPCK mRNA tended to increase from calving to 56 DIM. Pyruvate carboxylase mRNA increased more than twofold at calving. The data indicated adaptation to lactation for gluconeogenic enzymes that is not matched in direction and magnitude by changes in mRNA for urea cycle enzymes. Feeding additional protein, as RUP, failed to induce mRNA for key enzymes in gluconeogenesis or ureagenesis.

Animal Feed↗

PubMed2000

Dietary psyllium increases expression of ileal apical sodium-dependent bile acid transporter mRNA coordinately with dose-responsive changes in bile acid metabolism in rats.

Psyllium (PSY), a type of dietary fiber containing mainly soluble components, has been shown to decrease serum cholesterol concentrations in several species; however, mechanisms involved are not clearly defined. Four groups of 10 rats were fed semipurified diets containing 10% dietary fiber from cellulose and/or PSY for 21 d. Increasing levels of PSY were fed (0,3.33, 6.67 and 10% PSY) with the remaining 10% made up with cellulose. Liver cholesterol, cholesterol 7alpha-hydroxylase (CYP7A) activity and mRNA, 3-hydroxy-3-methylglutaryl CoA reductase (HMGR) mRNA, ileal apical sodium-dependent bile acid transporter (ASBT) mRNA, fecal bile acids and total steroids, and intestinal bile acid content were measured. All variables responded in a dose-dependent manner to PSY in the diet. Total liver cholesterol content was significantly reduced in all groups fed PSY compared to cellulose-fed controls [138(a), 105(b), 105(b) and 93(c) micromol (SEM = 4.2) for 0, 3.33, 6.67 and 10% PSY, respectively]. Activity of CYP7A was significantly greater in all groups fed PSY compared to the cellulose-fed controls [6.36(c), 16.92(b), 15.28(b) and 20.37(a) pmol x min(-1) x mg protein(-1) (SEM = 3.19) for 0, 3.33, 6.67 and 10% PSY, respectively]. These differences in CYP7A activity were similar to differences in CYP7A, HMGR and ASBT mRNA levels. Fecal bile acid and total steroid excretion as well as total intestinal bile acids were significantly greater in rats fed PSY-containing diets compared to 0% PSY-fed rats. These results suggest that the reduction in liver cholesterol involves modulating the size and composition of the bile acid pool via regulation of ileal ASBT, CYP7A and HMGR mRNA levels.

Animals↗

PubMed2000

Impact of dietary protein amount and rumen undegradability on intake, peripartum liver triglyceride, plasma metabolites, and milk production in transition dairy cattle.

Feeding strategies of transition dairy cows contribute to the risk factors associated with metabolic disorders that limit production in the ensuing lactation. To investigate the effects of prepartum dietary crude protein (CP) concentration and amount of rumen-undegradable protein (RUP) on postpartum health and production, 44 multiparous Holstein cows were blocked by expected calving date and assigned to one of four isoenergetic prepartum rations beginning 28 d prior to expected calving date. Prepartum rations were: 12% CP and 26% RUP, 16% CP and 26% RUP, 16% CP and 33% RUP, or 16% CP and 40% RUP on a dry matter basis. All cows were fed the same postpartum diet (18% CP, 40% RUP) from 1 to 56 d in milk (DIM). Prepartum dry matter intake (DMI) was not different among dietary treatments. Mean postpartum intakes (kg/d) were higher through 56 DIM (P<0.05) for cows fed the 12% CP:26% RUP diet prepartum compared with any of the 16% CP diets (21.8 vs. 19.8, 18.6 and 18.6; 12% CP:26% RUP vs. 16% CP:26% RUP, 16% CP:33% RUP and 16% CP:40% RUP). There was a DIM x prepartum diet interaction (P<0.05) with the greatest effect of the 12% CP:26% RUP diet evident during the first 35 DIM. Cows fed the 12% CP:26% diet during the transition period tended to produce more milk (kg/d) (P = 0.08) than did cows fed any of the 16% CP diets (40.8 versus 37.8, 38.7, and 37.4; 12% CP:26% RUP vs. 16% CP:26% RUP, 16% CP:33% RUP, and 16% CP:40% RUP). Additional protein (12 vs. 16% CP) in the prepartum diet tended to decrease milk protein (P = 0.10) and milk fat yield (P = 0.08) but did not alter percent milk fat, percent milk protein, or MUN. Liver triglyceride (TG) expressed as milligrams of TG per microgram of DNA or percentage of dry matter (DM) on d -28, -14, +1, +28, and +56 relative to calving were not significantly different among treatments. Maximal (P<0.05) infiltration of TG in liver was observed on +1 d when expressed as a percentage of DM and on +28 d when expressed as milligrams of TG per microgram of DNA. Plasma glucose, calcium, urea nitrogen, beta-hydroxybutyrate, and nonesterified fatty acids were not different (P<0.05) among treatments. The data indicate carryover effects of prepartum dietary protein on postpartum intake and milk production, pointing to beneficial effects of maintaining dietary protein for dairy cows in late gestation at 12% CP.

Animal Nutritional Physiological Phenomena↗

PubMed2000

Changes in mRNA expression for gluconeogenic enzymes in liver of dairy cattle during the transition to lactation.

The objective of this study was to profile phosphoenolpyruvate carboxykinase (PEPCK) and pyruvate carboxylase (PC) mRNA expression in the liver of dairy cattle during the peripartum transition and determine changes in abundance of these mRNA in response to protein fed during the prepartum period. Thirty-eight multiparous Holstein cows were fed diets containing either 12% crude protein (CP) and 26% rumen undegradable protein (RUP), 16% CP and 26% RUP, 16% CP and 33% RUP, or 16% CP and 40% RUP on a dry-matter basis beginning 28 d before expected calving. After calving, all cows were fed a common diet through 56 d in milk (DIM). Northern analysis of RNA from liver biopsy samples obtained on days -28, -14, +1, +28, and +56 relative to calving indicated that PC and PEPCK mRNA expression were responsive to onset of lactation but not to prepartum protein or RUP concentration. Abundance of PEPCK mRNA was similar at -28, -14, and +1 DIM but was elevated by +28 and +56 DIM relative to precalving levels. Liver PC mRNA abundance was elevated on +1 DIM, remained elevated through 28 DIM, and declined to precalving levels by 56 DIM. The activity of PC enzyme was correlated (r2 = 0.89) with PC mRNA abundance. The data demonstrate increased abundance of PC mRNA during the early transition period followed by increased abundance of PEPCK mRNA during the postpartum period and suggest increased potential metabolism of lactate, pyruvate, and amino acids that contribute to the liver pyruvate pool.

Animals↗

PubMed2000

Impact of dietary rumen undegradable protein and rumen-protected choline on intake, peripartum liver triacylglyceride, plasma metabolites and milk production in transition dairy cows.

The objectives of the present study were to determine the effects of rumen undegradable protein (RUP) level of prepartum diets, the supplementation of a rumen-protected choline product, and their interactions on milk production, feed intake, body weight and condition, blood metabolites, and liver triacylglycerides in dairy cows. Rumen-protected choline (RPC) was fed with two levels of RUP to 48 multiparous Holstein cows in a 3 x 2 factorial arrangement of treatments. Beginning 28 d before expected calving, cows were fed 10% rumen degradable protein, either 0, 6, or 12 g/d of RPC as CapShure (Balchem Corp., Slate Hill, NY) and either 4.0 or 6.2% RUP. After calving and through 120 d of lactation, cows received a common diet and continued RPC as per their prepartum assignment. Prepartum dry matter intake (kg/d) was not affected by RPC or RUP. Postpartum intake decreased when 6.2% RUP was fed prepartum. Milk production to 56 d in milk was decreased when cows were fed 6.2% RUP prepartum. Milk protein (kg/d) decreased when additional RUP was fed prepartum. Cows fed RPC lost more weight during the study period and tended to lose more body condition. Plasma urea nitrogen levels in the prepartum period were reduced for cows fed 4.0% RUP prepartum. Mean liver triacylglyceride determined from samples obtained at -28, -14, +1, +28, and +56 d in milk was not affected by RPC, prepartum RUP, or their combinations. Feeding 12 g of RPC/d in conjunction with 4.0% RUP increased milk production, but feeding RPC with 6.2% RUP prepartum and through 56 d in milk decreased production. The data indicate that 6.2% RUP does not benefit close-up dry cows, and the response to RPC depends the RUP content of the prepartum diet.

Animal Nutritional Physiological Phenomena↗

PubMed1999

Select 3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitors vary in their ability to reduce egg yolk cholesterol levels in laying hens through alteration of hepatic cholesterol biosynthesis and plasma VLDL composition.

The inability to markedly attenuate cholesterol levels in chicken eggs has led to speculation that cholesterol is essential for yolk formation and that egg production would cease when yolk cholesterol deposition was inadequate for embryonic survival. However, this critical level hypothesis remains unproven. Here, we determine the relative responsiveness of laying hens to three select inhibitors of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGR), the rate-limiting enzyme of cholesterol biosynthesis. A control diet, either alone or supplemented with one of two dietary levels (0.03 or 0.06%) of atorvastatin, lovastatin, or simvastatin, was fed to White Leghorn hens for 5 wk. Liver cholesterol concentrations (mg/g tissue) were decreased (P </= 0.05) by each HMGR inhibitor; however, total liver cholesterol (mg) did not differ among treatments. Microsomal hepatic HMGR activities were increased one- to twofold in all HMGR inhibitor-treated groups, while HMGR mRNA levels were unaffected. Diameters of plasma VLDL particles, the main cholesterol-carrying yolk precursor macromolecules, were reduced (P </= 0.05) only in hens fed 0.06% atorvastatin, and the particles contained 38% less total cholesterol (P </= 0.05) than controls. Plasma total cholesterol concentrations were lowered (P </= 0.05) by both doses of atorvastatin (-56, -63%) and simvastatin (-36,-45%). Egg cholesterol contents were maximally reduced by 46% (P </= 0.05), 7% (P > 0.05), and 22% (P </= 0.05) in hens fed the 0.06% level of atorvastatin, lovastatin, and simvastatin, respectively, while overall egg production [-19% (P </= 0.05), +4% (P > 0.05), and -3% (P > 0.05)], was much less affected. We concluded that cholesterol per se may not be an obligatory component for yolk formation in chickens and, as such, may be amenable to further pharmacological manipulation

Animals↗

PubMed1998

Dietary psyllium increases fecal bile acid excretion, total steroid excretion and bile acid biosynthesis in rats.

Psyllium, a source of dietary fiber rich in soluble components results in lower serum cholesterol concentration in several species. Suggested mechanisms for the hypocholesterolemic effect include a greater excretion of fecal bile acids and total steroids, and up-regulation of bile acid biosynthesis. The activity of cholesterol 7alpha-hydroxylase (7alphaOHase), the rate limiting enzyme in bile acid biosynthesis, is higher in rats fed 5% psyllium. Whether this higher activity corresponds to an increase in mRNA levels has not been determined. Four groups of 10 rats were fed a semipurified diet containing 5% cellulose (CEL; control), 5% cellulose plus 1% cholic acid (CCA), 5% cellulose plus 2% cholestyramine (CHY) or 5% psyllium hydrocolloid (PSY) for 3 wk. Liver cholesterol concentration, fecal bile acid and total steroid excretion, 7alphaOHase activity and 7alphaOHase mRNA levels were measured. Liver cholesterol content in rats fed CCA was significantly higher than in all other groups. Rats fed CHY and PSY had significantly lower liver cholesterol content than those fed CEL. Total fecal steroid and bile acid excretions were significantly greater in rats fed CCA, CHY and PSY than in those fed CEL. Activities and mRNA levels of 7alphaOHase in rats fed CHY and PSY were significantly higher than in rats fed CEL or CCA. These data indicate that feeding psyllium to rats increases fecal bile acid and total steroid excretion as well as 7alphaOHase activity and 7alphaOHase mRNA levels.

Animals↗

PubMed1997

Chronic and transitional regulation of gluconeogenesis and glyconeogenesis by insulin and glucagon in neonatal calf hepatocytes.

Milk-fed calves were used as a source of hepatocytes to establish monlayers to test the effects of chronic (41 h) incubation with no hormone, 100 nM insulin, or 100 nM glucagon on gluconeogenesis (glucose retained as glycogen plus glucose released into the medium) from 2.5 mM [2-(14)C]propionate and 2.0 mM [U-14C]lactate (1.0 mM lactate plus 1.0 mM pyruvate) during a subsequent 3-h (acute) incubation. Media for acute incubations contained no hormone, 0 or 10 nM insulin, or 0, 1, 10, or 100 mM glucagon. Chronic glucagon increased gluconeogenesis from propionate and glyconeogenesis from propionate and lactate compared with chronic exposure to medium with no hormone. A chronic glucagon x acute hormone interaction was manifested as an augmented response to acute glucagon on gluconeogenesis from propionate; a similar potentiation was not evident for gluconeogenesis from lactate. The concentration of glucagon required to acutely stimulate gluconeogenesis was increased by prior incubation with glucagon. Acute glucagon decreased the flux of glucose retained as glycogen regardless of chronic hormone treatment. Chronic incubation with insulin diminished the stimulatory effects of glucagon on gluconeogenesis from lactate. Chronic incubation with insulin did not alter the sensitivity or responsiveness at acute glucagon of gluconeogenesis from propionate. The data demonstrate persisting changes that favor increased basal gluconeogenesis from propionate with chronically elevated glucagon coupled to an increased capacity to respond acutely to glucagon and opposing chronic actions of insulin on lactate metabolism. These data suggest that insulin and glucagon target separate pathways that are unique to metabolism of propionate and lactate.

Animals↗

PubMed1997

Effects of fatty acids and hormones on fatty acid metabolism and gluconeogenesis in bovine hepatocytes.

Primary cultures of hepatocytes were used to study the effects of extracellular oleate concentration and hormones on fatty acid metabolism and gluconeogenesis. Rates of oleate uptake and oxidation to acid-soluble products varied linearly as oleate concentrations increased (0.1 to 2 mM), but rates of triglyceride accumulation varied quadratically. Insulin increased the proportion of oleate that was esterified by 22% without affecting the formation of acid-soluble products. Cells incubated with 2 mM [1-(14)C]oleate for 24 h eliminated 9.6% of the labeled intracellular lipid as acid-soluble products in the following 24 h when no oleate was present during depletion and eliminated 7.7% when 2 mM oleate was present. Insulin reduced labeled triglyceride depletion by 49%. Gluconeogenesis from [2-(14)C] propionate was depressed by 24%, and formation of acid-soluble products was increased by 46% in cells infiltrated with lipid because of previous exposure to 2 mM oleate for 45 h. Rates of gluconeogenesis from propionate were reduced 23% when 2 mM oleate was present during the 3-h period that gluconeogenesis was measured, and the effect was not modified by lipid infiltration. Lipid infiltration influenced hepatic function, and insulin regulated hepatic triglyceride concentration.

Animals↗

PubMed1996

Porcine somatotrophin differentially down-regulates expression of the GLUT4 and fatty acid synthase genes in pig adipose tissue.

The present study was conducted to determine whether porcine somatotropin (pST) differentially regulates expression of the GLUT4 and fatty acid synthase (FAS) genes in pig adipose tissue. Three different experiments were conducted in which pigs were treated daily with different doses of pST for different time periods (7 or 14 d and from 60 to 90 kg of body wt). In these experiments, pST significantly and consistently decreased FAS mRNA levels (80%, 66% and 85%, respectively); however, GLUT4 mRNA was not affected by pST in two of the three experiments, and in the one showing an effect (Experiment 2), the decrease was less than observed for FAS (44%). Because of these results, we conducted subsequent experiments to see if the effects of pST on glucose metabolism in cultured pig adipose tissue (48 h) differed when glucose concentrations were changed from 1 to 5 mmol/L. These studies revealed that the antagonistic effect of pST on insulin action was more potent when glucose transport was saturated (5 mmol/L) than when glucose concentration limited glucose entry into the cell (1 mmol/L). In summary, these results suggest that the effects of pST on glucose transport in pig adipocytes are secondary to changes elicited by the hormone on intracellular glucose use for lipogenesis. When considered in the context of the decrease previously observed in glucose transport in pig adipocytes, the findings reported herein suggest that pST acts to decrease GLUT4 protein activity and/or distribution between the plasma membrane and the intracellular pool with little alteration in GLUT4 gene expression or total cell GLUT4 protein.

Actins↗