Stimulation of lipogenesis in bovine adipose tissue by insulin and insulin-like growth factor

The present study was undertaken to determine if insulin and insulin-like growth factor 1 (IGF-1) stimulated lipogenesis in bovine adipose tissue and determine the effects of insulin on lipogenic capacity in adipose tissue cultured for 48 h. In contrast to previous studies, insulin markedly stimulated lipogenesis in short-term (2 h) incubations. The stimulation of lipogenesis by insulin was dependent upon the source of bovine serum albumin used in the buffer. Insulin-like growth factor 1 also stimulated lipogenesis; however, the potency was 80- to 100-fold lower than for insulin. Lipogenic capacity was decreased approximately 75% after 48 h of culture in the absence of insulin. When insulin was present in the culture medium, the reduction in lipogenic capacity was attenuated in a dose-dependent manner. However, insulin alone did not totally maintain lipogenic capacity after 48 h. In contrast, inclusion of hydrocortisone (HC; 50 ng/ml) and insulin (10 ng/ml) in the medium completely prevented the decline in lipogenic capacity of cultured bovine adipose tissue. In summary, these results indicate that bovine adipocytes are quite sensitive to insulin in short-term in vitro incubations and that insulin plays a predominant role in maintenance of lipogenic capacity of bovine adipose tissue during culture. Furthermore, the marked potentiation of insulin's effects of lipogenesis after 48 h of culture by HC suggests that the glucocorticoid is involved in regulation of insulin receptor number and(or) other cellular proteins (e.g., enzymes) which are important for lipogenesis to occur.     

The relationship between insulin-like growth factor-1, growth hormone, thyroid hormones and insulin in chickens selected for growth

The concentrations of circulating insulin-like growth factor I, growth hormone, insulin and thyroid hormones were measured in broilers selected for an increase in growth, broilers in which selection pressure was relaxed and in White Leghorns. Growth hormone levels increased in all lines between 3 and 4 weeks of age followed by a decline to adult levels. The lines with the slowest rate of growth had the highest growth hormone concentrations. Insulin-like growth factor I concentrations increased significantly in all three lines of birds during the 10 weeks of study and was significantly correlated with the increase in body weight. There were no consistent differences in plasma IGF-1 levels between the lines. Thyroxine levels increased consistently throughout the study but the levels of triiodothyronine decreased between 5 and 6 weeks of age in all lines. There were no consistent changes in plasma insulin levels. The highest rate of growth in these animals is accompanied by an increase in growth hormone concentration followed by an increase in plasma IGF-1. However, despite differences in plasma growth hormone, plasma concentrations of IGF-1 are not different between lines and are not related to between line differences in growth rate.     

Effects of ovine growth hormone and other anterior pituitary hormones on lipolysis of rat and ovine adipose tissue in vitro

Ovine growth hormone ( oGH ) was tested for its effects on lipolysis of rat and ovine adipose tissue in vitro. Ovine growth hormone at 1, 5 and 25 micrograms/ml stimulated lipolysis (P less than .05) of chopped rat adipose tissue and isolated rat adipocytes incubated in the presence of 100 mU/ml adenosine deaminase and .2 micrograms/ml dexamethasone, but had no effect on lipolysis of chopped ovine adipose tissue or isolated ovine adipocytes. Isoproterenol, a beta-adrenergic agonist, stimulated lipolysis (P less than .05) of both rat and ovine adipose tissue. Contaminants of the oGH preparation used were examined for lipolytic effects. Thyroid-stimulating hormone (TSH), luteinizing hormone (LH) and adrenocorticotropic hormone (ACTH) content in oGH were measured by radioimmunoassay. When quantities of these hormones contaminating 5 and 25 micrograms oGH were tested for lipolysis in rat adipose tissue, the TSH contamination could account for some (30%) of the lipolysis observed with oGH , while the other hormones had no effect. Also, preincubation of oGH with anti-GH, but not with anti-TSH or anti-LH, removed the principle in oGH responsible for the lipolytic effect on rat adipose tissue.(ABSTRACT TRUNCATED AT 250 WORDS)     

Patterns of gene expression in pig adipose tissue: insulin-like growth factor system proteins, neuropeptide Y (NPY), NPY receptors, neurotrophic factors and other secreted factors

Although cDNA microarray studies have examined gene expression in human and rodent adipose tissue, only one microarray study of adipose tissue from growing pigs has been reported. Total RNA was collected at slaughter from outer subcutaneous adipose tissue (OSQ) and middle subcutaneous adipose tissue (MSQ) from gilts at 90, 150, and 210 d (n=5 age(-1)). Dye labeled cDNA probes were hybridized to custom porcine microarrays (70-mer oligonucleotides). Gene expression of insulin-like growth factor binding proteins (IGFBPs), hormones, growth factors, neuropeptide Y (NPY) receptors (NPYRs) and other receptors in OSQ and MSQ changed little with age in growing pigs. Distinct patterns of relative gene expression were evident within NPYR and IGFBP family members in adipose tissue from growing pigs. Relative gene expression levels of NPY2R, NPY4R and angiopoietin 2 (ANG-2) distinguished OSQ and MSQ depots in growing pigs. We demonstrated, for the first time, the expression of IGFBP-7, IGFBP-5, NPY1R, NPY2R, NPY, connective tissue growth factor (CTGF), brain-derived neurotrophic factor (BDNF) and ciliary neurotrophic factor (CNTF) genes in pig adipose tissue with microarray and RT-PCR assays. Furthermore, adipose tissue CTGF gene expression was upregulated while NPY and NPY2R gene expression were significantly down regulated by age. These studies demonstrate that expression of neuropeptides and neurotrophic factors in pig adipose tissue may be involved in regulation of leptin secretion. Many other regulatory factors were not influenced by age in growing pigs but may be influenced by location or depot.     

IGF-1和IGFBP-3在5个地方猪种血液中含量及其与猪肉质性状相关性分析

为研究胰岛素样生长因子-1 (IGF-1)和胰岛素样生长因子结合蛋白-3 (IGFBP-3)在血液中的含量及其与胴体和肉质表型指标的相关性,采用ELISA技术检测了丫杈猪、乌金猪、成华猪、雅南猪和藏猪血液中IGF-1和IGFBP-3的含量,同时测定了肌纤维直径和眼肌面积.结果显示:藏猪肌纤维直径最小,且显著低于丫杈猪、...     

Immunohistochemical detection of components of the insulin-like growth factor system during skeletal muscle growth in the pig

The insulin-like growth factor (IGF) system plays an important role in postnatal somatic and skeletal muscle growth in pigs. There is little information on the occurrence and distribution of components of the IGF system in postnatal porcine skeletal muscle. IGF-I, IGF receptor 1 (IGF1R) and the IGF-binding proteins IGFBP-1 and -3 in longissimus dorsi and triceps brachii were localized in muscle biopsies from 12 commercially crossbred pigs aged from 28 to 199 days as well as from the sire generation, by immunohistochemistry. Plasma IGF-I concentrations were also determined using radio-immunoassays. Unlike other species, IGF-I was localized in porcine skeletal muscle fibres. Staining intensity correlated with the highest plasma IGF-I levels and phases of intensive muscle growth from the 11th to 22nd week. The pattern of IGF1R immunostaining, which was strong, correlated with that of IGF-I, IGF1R was also localized in endomysial tissues. IGFBP-1 was not detected within muscle fibres, but was found in the endomysium and vessel walls, while IGFBP-3 was localized with IGF-1 and its receptor. Higher magnification revealed that IGF1R, IGFBP-3 and probably IGF-I appeared in the tubular system. Inhibitory as well as stimulating controls of IGFBP-1 and -3 on IGF functions are discussed, which may maintain a balance between autocrine growth promoting activities of IGF-I and IGF1R.     

Measurements of growth hormone and insulin-like growth factor 1 in cats with diabetes mellitus

Serum concentrations of insulin-like growth factor 1 (IGF-1) and growth hormone were measured in 25 cats with untreated diabetes mellitus (11 of which were used for follow-up measurements, one to three, four to eight, nine to 12 and 13 to 16 weeks after their treatment with insulin began), 14 diabetic cats that had previously been treated with insulin, and seven diabetic cats that also had hypersomatotropism, two of which had not previously been treated with insulin; 18 healthy cats were used as controls. In the untreated diabetic cats the concentration of IGF-1 ranged from 13.0 to 433.0 ng/ml (median 170.5 ng/ml), which was significantly lower than the concentrations in the control cats (196.0 to 791.0 ng/ml, median 452.0 ng/ml). Their IGF-1 concentrations increased significantly when they were treated with insulin and after four to eight weeks were not different from those in the control cats. In the diabetic cats that had previously been treated with insulin the IGF-1 concentrations were 33.0 to 476.0 ng/ml (median 316.0 ng/ml), which was significantly lower than the concentrations in the control cats, but significantly higher than in the untreated diabetic cats. The IGF-1 concentrations in the two previously untreated diabetic cats with hypersomatotropism were low and low-normal but increased markedly after treatment with insulin. In the five previously treated cats with hypersomatotropism the concentration of IGF-1 was above the normal range. The concentrations of growth hormone in the treated and untreated diabetic cats without hypersomatotropisms were not significantly different and there was an overlap in its concentrations in the diabetic cats with and without hypersomatotropism.     

Differentiation of adipose tissue and muscle in hypophysectomized pig fetuses

In the present study, fetuses were hypophysectomized (hypox) in utero on d 72 to 74 of gestation with an electrical cauterizing needle. One to six successfully hypox fetuses were removed on d 110 of gestation from each of five gilts. Subcutaneous adipose tissue samples and semitendinosus muscles were obtained from the hypox fetuses and an equal number of control fetuses. Body weights of control fetuses (n = 15; mean +/- SE, 1,195 +/- 33 g) were similar to weights of hypox fetuses (n = 15; 1,179 +/- 67 g). Fat cell size in the middle subcutaneous layer of adipose tissue was increased in hypox fetuses (P less than .01) compared with control fetuses. The number of obvious fat cell clusters (outer layer) in lipid stained sections was reduced (P less than .01) by 50% in hypox fetuses. Histochemical reactions for glucose-6-phosphate dehydrogenase, esterase and lipoprotein lipase (LPL) activities in middle layer cell clusters were considerably enhanced in sections from hypox fetuses compared with sections from controls. Quantitative analysis of percent light transmittance (Zeiss photometer) through LPL-stained cell clusters indicated an increase (P less than .001) in LPL staining in sections from hypox fetuses when compared with sections from control fetuses. Transverse muscle sections (cryostat) from hypox fetuses failed to show normal patterns (as seen in control muscles) of reactions for acid ATPase, malate dehydrogenase (NAD-dependent), NADH-TR and alpha-glycerol phosphate dehydrogenase (without NAD). The number of muscle fibers that were stained for these enzymes was greatly reduced in hypox fetuses compared with control fetuses. The number of lipid positive fibers was also reduced in hypox fetuses compared with control fetuses.(ABSTRACT TRUNCATED AT 250 WORDS)     

The ovarian insulin and insulin-like growth factor system with an emphasis on domestic animals

Insulin and insulin-like growth factors (IGFs) have direct effects on cultured ovarian cells. These effects include stimulation of granulosa cell mitogenesis, granulosa and luteal cell progesterone production, and thecal cell androgen production and appear similar among species. However, species differences exist with regard to insulin and IGF-I effects on granulosa cell estradiol production. In addition to endocrine effects of insulin and IGFs, IGFs are produced by granulosa, thecal, and luteal cells, allowing for an intraovarian autocrine and paracrine system. Granulosa, thecal, and luteal cells contain receptors for insulin and IGFs, and these receptors appear to mediate the effects of insulin and IGFs. Adding to the complexity of the regulatory role of IGFs is the presence of IGF-binding proteins (IGFBPs) within the ovary. These IGFBPs are produced by granulosa, thecal, and luteal cells, and their production is hormonally regulated. Evidence for a coherent mechanism by which insulin, IGFs, and IGFBPs interact and regulate ovarian function in vivo has yet to be found.     

Influence of nutritional deprivation on insulin-like growth factor I, somatotropin, and metabolic hormones in swine

Although growth hormone (GH) is a primary stimulus for the synthesis of insulin-like growth factor I (IGF-I), other factors such as nutritional status, insulin, and thyroid hormones are important modulators of circulating IGF-I levels. Thus, the effects of feed restriction and subsequent refeeding on plasma levels of IGF-I, GH, insulin, and thyroid hormones were studied in swine. Despite an elevation in plasma GH levels after 48 h of feed restriction, circulating IGF-I levels were decreased by 53% (P less than .05). Plasma triiodothyronine (T3) and insulin were lower (P less than .05) within 24 h after the feed restriction began, whereas thyroxine (T4) did not decrease until 48 h after removal of feed. Blood glucose levels remained unchanged throughout the experiment. Refeeding after the 48-h fast was associated with a decline (P less than .05) in circulating GH levels within 2 h, concomitant with an elevation (P less than .05) in plasma insulin and T3. Refeeding fasted pigs was associated with an increase (P less than .05) in plasma IGF-I; however, levels still had not returned to prefast concentrations within 24 h after refeeding. These data indicate that the GH-IGF-I axis becomes uncoupled during nutritional restriction in swine and that inadequate nutrient supply may limit the expression of the anabolic effects of GH.     

Regulation of secondary follicle growth by theca cells and insulin-like growth factor 1

Ovaries contain follicles at various stages of development, including primordial, primary, secondary, antral and Graafian follicles. Although the growth of these follicles is controlled to maintain regular ovulation, the mechanism through which this occurs remains unclear. In our study, we found that the growth rate of cultured secondary follicles separated from mice ovaries differed between follicles. After 4 days of culture, the size of some secondary follicles was markedly increased, while that of others had either slightly increased, remained unchanged or shrunk. We compared the expression levels of growth factors between these secondary follicles and found that the growth rate of cultured secondary follicles correlated with the expression level of insulin-like growth factor 1 (Igf1) mRNA. Igf1 mRNA expression level in secondary follicles containing theca cells was higher than that in secondary follicles without theca cells, and the granulosa cell proliferation around follicles containing theca cells was increased. Furthermore, an IGF1 inhibitor also inhibited the granulosa cell proliferation, and administration of IGF1 to secondary follicles without growth promoted granulosa cell proliferation. These results indicated that the theca cells of secondary follicles induced the expression of IGF1 and promoted the follicle growth.     

Effect of pituitary somatotropin injections on plasma insulin-like growth factor I and somatotropin profiles in growing heifers

Effects of daily injections of pituitary-derived bovine somatotropin (bST) for 6 wk were evaluated in 10 growing heifers and compared to 9 placebo-treated control animals. Bovine somatotropin was injected at 50 micrograms/kg BW each day. Body weight and growth, plasma concentrations of insulin-like growth factor I (IGF-I) and somatotropin (ST) were assessed. To measure plasma concentrations of IGF-I, we validated a RIA in which bovine plasma samples were extracted with acid-ethanol, a method that resulted in greater than 90% recovery of IGF-I. Average daily gain was similar during the first 4 wk of the experiment in both control and bST-treated groups; however, at the end of the experimental period (wk 4 and 6) ADG was greater (P less than .05) in bST-treated heifers (1.24 +/- .21 kg/d vs .75 +/- .25 kg/d). Plasma IGF-I from wk 2 to wk 6 were increased in bST-treated animals (452 +/- 97 ng/ml at wk 2; 683 +/- 106 ng/ml at wk 6) compared with controls (293 +/- 62 ng/ml at wk 2 (P less than .01) and 293 +/- 115 ng/ml at wk 6 (P less than .001). Moreover, ADG over the 6-wk experimental period was correlated with mean IGF-I concentrations determined over the same period (r = .55; P less than .01). As expected, mean plasma ST concentrations were increased in bST-injected animals from wk 1 to 6. Gel chromatographic profiles of bovine plasma exhibit a 150,000 molecular weight ST-dependent binding protein-IGF-I complex and a 30,000 molecular weight non-ST-dependent complex. This study validates a method for measuring IGF-I in cattle, and shows a positive relationship among IGF-I and ADG after ST treatment. No correlation, however, was found between plasma ST and growth performance.     

Stimulation of lipogenesis by insulin in swine adipose tissue: antagonism by porcine growth hormone

The effects of physiological (1, 10 ng/ml) and pharmacological (1,000 ng/ml) concentrations of insulin (INS) and porcine growth hormone (pGH) on lipid metabolism were determined in short-term (2 h) and long-term (26, 50 h) incubations of swine adipose tissue. The short-term effects of three different commercial sources of bovine serum albumin (BSA) on adipose tissue metabolism were also evaluated. Two of the three BSA preparations were found to be unsuitable for inclusion in the short-term incubation buffer because they caused a stimulation of lipid synthesis in adipose tissue and masked the stimulatory effects of insulin. Physiological concentrations of insulin stimulated glucose metabolism in 2-h incubations by 100% in adipose tissue from 80-kg swine. After a 26-h incubation period, INS maintained rates of glucose metabolism at levels comparable to maximally stimulated rates in fresh tissue. Insulin also enhanced glucose metabolism following 50-h incubations; however, rates were less than for 2- or 26-h incubations. Glucose metabolism was also stimulated in adipose tissue from 127-kg swine when incubated for 2 h with INS; however, INS responsiveness declined with increasing body weight. Lipogenesis and glucose oxidation were partially maintained by INS using tissue from the heavier swine. A pharmacological but not physiological concentration of pGH stimulated glucose metabolism in short-term incubations by 50% in adipose tissue from 80-kg swine, and by 10% in adipose tissue from 127-kg swine. Long-term culture of adipose tissue in the presence of pGH had no effect on glucose metabolism. Physiological levels of pGH directly antagonized the stimulation of glucose metabolism by INS in short- and long-term incubations. In summary, these results are the first to establish that swine adipose tissue is quite sensitive to insulin and that pGH directly antagonizes insulin action.     

Effect of fat supplementation on leptin, insulin-like growth factor I, growth hormone, and insulin in cattle

We investigated the effect of fat supplementation on plasma levels of hormones related to metabolism, with special attention to leptin, in cows in early lactation and in feedlot steers. In experiment 1, 34 lactating cows received no fat or else 0.5 or 1.0 kg of partially hydrogenated oil per day in addition to their basal diet from day 20 before the expected calving date to day 70 postpartum. In experiment 2, part of the corn in the basal concentrate was replaced with 0.7 kg of the same oil such that the diets were isocaloric; 18 cows received the fat-substituted diet and 18 a control diet from day 20 before the expected calving date to day 75 postpartum. In experiment 3, calcium salts of fatty acids were added to the basal diet of 14 feedlot steers for 80 d; another 14 steers received a control diet. The basal plasma levels of leptin were higher in the cows than in the steers. Dietary fat supplementation did not affect the leptin levels in the lactating cows but lowered the levels in the feedlot steers despite greater energy intake and body fatness (body weight) in the steers receiving the supplement than in those receiving the control diet. The levels of insulin-like growth factor I and insulin were decreased with dietary fat supplementation in the lactating cows but were unaffected in the steers, suggesting that responses to fat ingestion depend on the physiological state of the animal, including age and sex. Finally, no effects of supplementary fat on the level of growth hormone were demonstrated in any of the models.     

Effects of peroral insulin and glucose on circulating insulin-like growth factor-I, its binding proteins and thyroid hormones in neonatal calves

There is disagreement in the literature about the ability of neonatal calves to absorb perorally administered insulin. This study evaluated the absorption of a bolus of insulin administered alone or with an energy souce and its effects on the circulating insulin-like growth factor system and thyroid hormones in newborn Holstein-Friesian calves. Within 1 h of dosing, mean serum insulin and triiodothyronine (T3) concentrations had increased considerably, whether the insulin was applied alone (n = 4) or together with glucose (n = 4), accompanied by marked hypoglycemia. No significant changes were observed in control calves (n = 4) given the vehicle solution. Increased serum glucose and T3 concentrations with no change in insulinemia occurred in a 4th group of calves given glucose alone. At 32 h of age and after 3 meals of colostrum there were no differences in glycemia, insulinemia, or proteinemia among the 4 groups of calves examined. Mean serum insulin-like growth factor-I (IGF-I) tended to decrease over this period in the control group. The decrease was more pronounced in the insulin-treated group but absent in both groups that received glucose. These differences were associated with equivalent differences in abundance of the 40-45K IGF-binding protein-3 (IGFBP-3); however, lower molecular mass IGFBPs were not affected. The results show that a pharmacological peroral dose of insulin can lead to rapid systemic alterations in the IGF/IGFBP system in neonatal calves that can be modified by simultaneous administration of a small energy supply in the form of glucose.