Effects of recombinant bovine somatotropin (rbST) and season on plasma and milk insulin-like growth factors I (IGF-I) and II (IGF-II) in lactating dairy cows

During two studies, effects of recombinant bovine somatotropin (rbST) on plasma and milk IGF's in cows adapted to summer (S; 12 cows) or winter (W; 12 cows) conditions were evaluated. Each study consisted of on-farm periods (30 days) followed by climatology chamber periods (CC; 30 days). Cows were given daily injections of rbST, Sometribove, USAN (25mg/day; 6 cows each study) or saline (control; 6 cows each study). During on-farm periods, blood and milk (am and pm) samples were collected once weekly. During CC periods, blood samples were collected every 2 days and milk samples (am and pm) were collected daily. Plasma IGF-I and IGF-II were increased in cows treated with rbST. A pronounced seasonal pattern in basal and rbST-stimulated plasma IGF-I but not IGF-II was detected. Higher basal and rbST-stimulated plasma IGF-I concentrations in S occurred despite large decreases in feed intake and energy balance. Milk IGF-I and IGF-II was not affected by rbST treatment or season. Although milk IGF-I and IGF-II concentrations were unaffected by rbST treatment, total IGF-output increased due to increased milk yield. The observed seasonal patterns in plasma IGF-I may be indicative of seasonal differences in the coupling of the somatotropin-IGF axis. In particular, we failed to detect an uncoupling of the somatotropin-IGF-I axis in S despite an induced negative energy balance during thermal stress.     

Effects of exogenous cortisol on the GH/IGF-I/IGFBP network in channel catfish

Glucocorticoids are known to hinder somatic growth in a number of vertebrate species. In order to better understand the mechanisms through which they may act in channel catfish, we examined the effects of feeding cortisol on the growth hormone (GH)/insulin-like growth factor-I (IGF-I)/IGF-binding protein (IGFBP) network. Fish (30.6 ± 3.0 g) were fed once daily for 4 weeks and treatments included: (1) High-cortisol (dietary cortisol provided at 400 mg/kg feed), (2) Low-cortisol (dietary cortisol provided at 200 mg/kg feed), and (3) Control (commercial catfish feed). Fish fed diets with cortisol weighed approximately 50% less than Controls. Feed intake was reduced by approximately 30% in both treatments of cortisol fed fish compared to Controls. A 20-kDa IGFBP was observed in plasma from High- and Low-treated fish while it was not detected in Control fish plasma. High-cortisol treatment increased pituitary GH mRNA expression approximately 10-fold while liver IGF-I mRNA expression was not different between cortisol-treated fish and Controls. Cortisol treatments decreased plasma levels of IGF-I. These data indicate that feeding cortisol for 4 weeks reduces weight gain, feed intake, and plasma levels of IGF-I and induces a 20-kDa IGFBP. One mechanism through which cortisol may impede growth of catfish is through an increase in a low molecular weight IGFBP which may lead to inhibitory effects on the action of IGF-I.     

Effects of fasting on circulating IGF-binding proteins, glucose, and cortisol in channel catfish (Ictalurus punctatus)

The effects of fasting on IGF-binding proteins (IGFBPs), glucose, and cortisol in channel catfish were examined. Fed fish (controls) were compared to 14-, 30-, and 45-day fasted fish and 45-day fasted fish refed for 15 additional days. Body length and weight changes, condition factor (CF), hepatosomatic index (HSI), and plasma glucose and cortisol were assessed to determine growth and metabolic status. Body length and growth rates were inhibited (P<0.05) after 14, 30, and 45 days of fasting. The 14-, 30-, and 45-day fasted fish exhibited hypoglycemia and reduced CF and HSI. Cortisol levels were increased (22.8 +/- 15.2 ng/ml versus 4.7 +/- 3.9 ng/ml) in 30-day fasted fish compared to fed controls (P<0.05). Associated with the increase in cortisol in fasted fish was a concomitant increase in plasma levels of a 20-kDa IGFBP through day 45. A 35- and a 45-kDa IGFBP were also identified but were similar between fed and unfed fish throughout the experiment. At the end of 15 days of refeeding, 20-kDa IGFBP, glucose, and cortisol levels were similar to fed controls. Refeeding also caused an increase in growth rates. These results suggest the existence of a catfish counter part to mammalian IGFBP-1, similar to lower molecular mass IGFBPs reported in other species of fish. These results also suggest that a 20-kDa IGFBP is upregulated during fasting-induced growth inhibition of channel catfish and provide additional evidence of the conserved nature of the IGF-IGFBP-growth axis in fish.     

Effects of recombinant bovine somatotropin on growth and abundance of mRNA for IGF-I and IGF-II in channel catfish (Ictalurus punctatus)

Research was conducted to examine growth rates, circulating concentrations of IGF-I, and mRNA abundance levels of IGF-I and IGF-II in channel catfish (Ictalurus punctatus) given recombinant bovine ST (rbST; Posilac, Monsanto Co., St. Louis MO). In the first study, juvenile catfish (5.5 +/- 0.5 g) were randomly assigned to one of three treatments: 1) sham-injected control (one needle puncture per week); 2) rbST (30 microg x g BW(-1) x wk(-1); Posilac); and 3) nonhandled control (control). At the end of the 6-wk study, the fish were weighed, measured for length, and G:F was determined. Compared with sham and control treatments, rbST-treated fish had 48% greater final BW, 14% greater total length, and 52% greater G:F (P < 0.001). In the second study, juvenile catfish (41.1 +/- 1.5 g) were assigned randomly to one of two treatments: 1) sham or 2) rbST. Eight fish per treatment were sampled on d 0, 1, 2, 7, 14, and 21 for blood, muscle, and liver. Relative expression of IGF-I and IGF-II mRNA was determined by real-time PCR and plasma concentrations of IGF-I were measured using a validated fluoroimmunoassay. Circulating concentrations of IGF-I were increased (37.9 +/- 5.5 vs. 22.0 +/- 6.6 ng/mL; P < 0.05) in rbST-injected fish compared with sham-injected controls by d 14. Liver IGF-I and IGF-II mRNA was increased 4.3-and 14.4-fold, respectively, by d 1 in rbST-injected fish compared with controls (P < 0.05); however, abundance of liver IGF-I and IGF-II mRNA did not differ from controls on d 0, 2, 7, 14, and 21. Abundance of muscle IGF-I and IGF-II mRNA did not differ in rbST-injected fish compared with controls throughout the study. Results of the first study demonstrated that rbST improves growth performance of channel catfish. Results of the second study showed that the growth-promoting effects of rbST were not mediated by the expression of IGF-I or IGF-II mRNA in the muscle. Instead, the results suggest that rbST promotes growth by stimulating plasma IGF-I release, possibly through its direct effect on the liver or on local tissues to synthesize IGF-I. The changes in mRNA abundance and plasma concentrations of IGF-I support the role of IGF-I in growth regulation of channel catfish.     

半胱胺和大豆黄酮对东北细毛羊部分组织IGF-I mRNA表达量的影响

为了探讨半胱胺和大豆黄酮对东北细毛羊肝脏、肌肉和皮肤组织中IGF-I mRNA表达量的影响,选取体重27～28 kg,5～6月龄健康的东北细毛羊育成羊21只,随机分成7组,每组3个重复,每个重复1只,采用实时荧光定量PCR法检测肝脏、肌肉和皮肤组织中IGF-I mRNA相对表达量.结果表明:(1)半胱胺和大豆黄酮均能显著提高东北细毛羊的生长性能(P<0.05),其中以7.5 mg/kg BW半胱胺组和7.5 mg/kg BW半胱胺+3 mg/kg BW大豆黄酮组效果较佳;(2)半胱胺和大豆黄酮对东北细毛羊肝脏和肌肉中IGF-I mRNA表达量有显著的提高作用(P<0.05),但各试验组之间差异不显著(P>0.05);(3)对皮肤中表达量的影响:对照组与添加3和6 mg/kg BW大豆黄酮组和7.5 mg/kg BW半胱胺+3 mg/kg BW大豆黄酮组之间差异显著(P<0.05),与添加7.5和15 mg/kg BW半胱胺组和15 mg/kg BW半胱胺+6 mg/kg BW大豆黄酬组之间差异不显著(P>0.05).由结果可知,半胱胺和大豆黄酮均能提高肝脏和肌肉中IGF-I mRNA的表达量,但协同应用并没有促使IGF-I mRNA表达量在肝脏、肌肉和皮肤组织中的明显提高,综合分析以添加3 mg/kg BW大豆黄唰效果最佳.     

Establishment of a time-resolved fluoroimmunoassay for measuring plasma insulin-like growth factor I (IGF-I) in fish: effect of fasting on plasma concentrations and tissue mRNA expression of IGF-I and growth hormone (GH) in channel catfish (Ictalurus punctatus)

A time-resolved fluoroimmunoassay (TR-FIA) was established and validated that allows for the determination of plasma concentrations of insulin-like growth factor I (IGF-I) in three domestically cultured fishes: channel catfish (Ictalurus punctatus), hybrid striped bass (Morone chrysopsxM. saxatilis), and rainbow trout (Oncorhynchus mykiss). Sensitivity of the assay was 0.20 ng/ml. Intra- and inter-assay coefficients of variation (CV) were <7 and <12%, respectively. Serial dilutions of plasma from each species were parallel to the standard curve. Recovery of IGF-I from spiked plasma samples was >90% for all three species of fishes. The IGF-I TR-FIA was biologically validated via its use to determine the effect of fasting on circulating IGF-I levels in channel catfish. Fasting-induced changes in plasma growth hormone (GH), hepatic IGF-I mRNA expression, and pituitary GH mRNA expression were also determined. Fasted channel catfish lost 5.6 and 15.6% body mass after 2 and 4 weeks of fasting, respectively. Plasma IGF-I concentrations were depressed (P<0.05) relative to fed controls following 2 and 4 weeks of fasting. Plasma GH concentrations were not different (P>0.05) in fasted fish after 2 weeks, but significantly increased (P<0.05) by 4 weeks of fasting. Hepatic IGF-I mRNA expression after 2 and 4 weeks of fasting was reduced (P<0.05) relative to fed controls. Pituitary GH mRNA expression was similar (P>0.05) between 2-week-fasted catfish and fed controls, but was increased (P<0.05) in 4-week-fasted catfish. The IGF-I TR-FIA was sensitive, accurate, and precise for all three species of fishes, and provided a low-cost, and non-radioisotopic method for quantifying plasma IGF-I levels in fed and fasted channel catfish.     

溶菌酶和抗生素对肉鸡组织IGF-Ⅰ mRNA的影响

选择144只1日龄艾维因肉鸡分为4组,对照组饲喂基础日粮,抗生素组饲喂添加抗生素(有效成分为杆菌肽锌和硫酸粘杆菌素)的基础日粮,2个溶菌酶组分别饲喂基础日粮添加溶菌酶150mg/kg和300mg/kg的日粮,每组6个重复,每个重复6只,于42日龄取样,采用RT-PCR方法测定肉鸡肝脏和胸肌IGF-ⅠmRNA的相对表达水平。试验结果表明:溶菌酶组较对照组可将肝脏IGF-ⅠmRNA水平分别提高12.5%和10.4%,但与对照组及抗生素组相比差异不显著(P0.05);各处理对胸肌IGF-ⅠmRNA水平有提高的趋势,但无显著差异(P0.05)。     

Effects of short- and long-term fasting on plasma and stomach ghrelin,and the growth hormone/insulin-like growth factor I axis in the tilapia, Oreochromis mossambicus

Ghrelin is a highly conserved peptide hormone secreted by the stomach, which is involved in the regulation of food intake and energy expenditure. Ghrelin stimulates growth hormone (GH) release, and increases appetite in a variety of mammalian and non-mammalian vertebrates, including several fish species. Studies were conducted to investigate the effect of feeding and fasting on plasma and stomach ghrelin, and the growth hormone/insulin-like growth factor I (IGF-I) axis in the Mozambique tilapia, a euryhaline teleost. No postprandial changes in plasma and stomach ghrelin levels or stomach ghrelin mRNA levels were observed. Plasma levels of GH, IGF-I and glucose all increased postprandially which agrees with the anabolic roles of these factors. Fasting for 4 and 8 d did not affect ghrelin levels in plasma or stomach. Plasma GH was elevated significantly after 4 and 8 d of fasting, while plasma IGF-I levels were reduced. Plasma ghrelin levels were elevated significantly after 2 and 4 wk of fasting, but no change was detected in stomach ghrelin mRNA levels. Four weeks of fasting did not affect plasma GH levels, although plasma IGF-I and glucose were reduced significantly, indicating that GH resistance exists during a prolonged nutrient deficit (catabolic state). These results indicate that ghrelin may not be acting as a meal-initiated signal in tilapia, although it may be acting as a long-term indicator of negative energy balance.     

Immunolocalization of insulin-like growth factor-I (IGF-I) and its receptors (IGF-IR) in the equine epididymis

Insulin-like growth factor plays a paracrine/autocrine role in regulating testicular function in the stallion, but its presence in the equine epididymis remains unknown. The aim of this study was to test the hypothesis that insulin-like growth factor-I (IGF-I) and IGF-I receptor (IGF-IR) are localized in the caput, corpus, and cauda of the epididymis in an age-dependent manner. Immediately after castration, epididymal tissue was fixed, paraffin-embedded, and processed for immunohistochemistry (IHC). Western blot was also performed using equine epididymal extracts to verify the specificity of the antibodies against IGF-I and IGF-IR. Immunolabeling of IGF-I was observed in the cytoplasm of principal and basal cells in the caput, corpus, and cauda at the pre-pubertal (3–7 months), pubertal (12–18 months), post-pubertal (2–4 years), and adult stages (4.5–8 years). Immunolabeling of IGF-IR was observed in the cytoplasm of principal cells in all regions of the epididymis in each age group. Immunolabeling of IGF-IR was also detected in the cytoplasm of basal cells from animals of all ages. Bands observed by Western blot corresponded to the molecular weights of IGF-I and IGF-IR, ~23 kDa and 95 kDa, respectively. These results suggest that IGF-I might function as an autocrine and/or paracrine factor during the development, maintenance and/or secretions of the stallion epididymis.     

日粮色氨酸水平对生长猪的氮利用效率、血浆类胰岛素生长因子-I、生长激素及胰岛素的影响

本试验旨在研究日粮色氨酸水平对生长猪的氮利用率以及对血浆类胰岛素生长因子-I(IGF-I)、生长激素(GH)、胰岛素浓度的影响。试验以4头大白去势公猪作为试验动物,以色氨酸含量分别为1.2、1.7、2.2和2.7g/kg的4种日粮为试验处理,按照4×4拉丁方进行试验设计。结果表明,色氨酸水平对干物质消化率、有机物消化率和氮消化率没有显著性影响(P>0.05)。氮沉积和氮的生物学利用率随着日粮色氨酸水平由1.2提高到1.7、2.2和2.7g/kg而显著提高(P<0.05),但是试验处理1.7、2.2和2.7g/kg之间的氮沉积和氮的生物学价值没有显著差异(P>0.05)。不同处理之间的血浆IGF-I、GH和胰岛素浓度没有显著差异(P>0.05)。日粮色氨酸水平和血浆IGF-I、GH或胰岛素浓度之间以及氮沉积和血浆IGF-I、GH或胰岛素浓度之间均无显著的相关关系(P>0.05)。综合以上结果,日粮色氨酸水平对生长猪的氮沉积具有显著影响,而对血浆IGF-I、GH和胰岛素没有显著影响。     

Evaluation of serum insulin-like growth factor binding proteins (IGFBP) in Angus cattle divergently selected for serum IGF-I concentration

Postweaning serum insulin-like growth factor-I (IGF-I) concentrations and serum IGF binding proteins (IGFBP) were investigated in 68 (1992 Fall-born) and 84 (1999 Fall-born) Angus cattle selected for either high or low serum IGF-I concentrations since 1989. Relative serum levels of IGFBP were determined by [¹²⁵I]IGF-I Western ligand blotting. IGFBP species of 38–42, 34, 30, and 24 kDa were identified. The 34 kDa species was identified as IGFBP-2 by immunoblot analysis. No significant line effects were observed for any of the IGFBP. In both 1992 and 1999, heifers had higher IGFBP-2 levels than bulls (P<0.0005). In 1992 calves, relative levels of the 38–42 and 24 kDa species were significantly correlated with serum IGF-I concentration. In 1999 calves, none of the IGFBP were correlated with serum IGF-I, although IGFBP-2 was negatively correlated with several measures of body weight. No significant line effects were observed for growth or serum IGF-I traits in 1992 calves. However, 1999 high line calves had higher serum IGF-I concentrations and body weights than low line calves (P<0.05). In both 1992 and 1999 calves, bulls had higher serum IGF-I concentrations and body weights than heifers (P<0.05). Thus, while selection for high versus low serum IGF-I concentrations has resulted in divergence between the selection lines and also in changes in body weights, it has not resulted in changes in serum IGFBP levels. Furthermore, circulating IGFBP-2 appears to be higher in heifers than in bulls, and also appears to be negatively correlated with body weights.     

Effects of EGF and IGF-I on in vitro Maturation and Cleavage of Ovine Oocytes

The study investigated the effects of epidermal growth factor(EGF) and insulin-like growth factor 1(IGF-I),alone or together,on the in vitro maturation and cleavage of ovine oocytes,aimed to optimize the in vitro maturation conditions for ovine oocytes.The results showed that the maturation and cleavage rates were 71.2% and 45.5% respectively when the medium was supplemented with 50 ng/mL EGF alone,which was significantly higher than other EGF supplemented groups (0,10,20,30,and 40 ng/mL) (P<0.05).The highest maturation and cleavage rates were 72.9% and 45.7% when the EGF concentration reached 100 ng/mL.The maturation and cleavage rates were 70.7% and 58.5% with 40 ng/mL IGF-I supplemented,which were significantly higher than other treatments (0,10,20,60,80,and 100 ng/mL) (P<0.05).The lowest maturation and cleavage rates were 38.8% and 20.0% when the IGF-I concentration reached 100 ng/mL (P<0.05).When 50 ng/mL EGF and 40 ng/mL IGF-I were used concomitantly,the maturation and cleavage rates were 85.6% and 61.0% respectively,which were significantly higher than the treatments with EGF or IGF-I alone (P<0.05).     

Potential role of G-protein-coupled receptor 30 (GPR30) in estradiol-17beta-stimulated IGF-I mRNA expression in bovine satellite cell cultures

Androgenic and estrogenic steroids enhance muscle growth in animals and humans. Estradiol-17beta (E2) and trenbolone acetate (TBA) (a synthetic testosterone analog) increased IGF-I mRNA expression in bovine muscle satellite cell (BSC) cultures. The goal of this study was to evaluate the mechanisms responsible for this increase by evaluating the effects of ICI 182 780 (an E2 receptor antagonist), flutamide (an androgen receptor inhibitor), G1 (a GPR30 agonist), and BSA-conjugated E2 on E2 and/or TBA-stimulated IGF-I mRNA expression in BSC cultures. Flutamide completely suppressed TBA-stimulated IGF-I mRNA expression in BSC cultures. ICI 182 780 did not suppress E2-stimulated IGF-I mRNA expression and 100nM ICI 182 780 enhanced (93%, p<0.05) IGF-I mRNA levels in BSC cultures. G1 (100nM) stimulated IGF-I mRNA expression (100%, p<0.05) but had no effect on proliferation in BSC cultures. E2-BSA, which cannot cross the cell membrane, stimulated IGF-I mRNA expression (approximately 100%, p<0.05) in BSC but even at extremely high concentrations had no effect on proliferation. In summary, our data indicate the E2-stimulation of proliferation and E2-stimulation of IGF-I mRNA expression in BSC cultures occur via different mechanisms. Our previous results showing that ICI 182 780 inhibited BSC proliferation and results of the current study showing lack of response to E2-BSA or G1 suggest that E2-stimulated proliferation in BSC cultures is mediated through classical estrogen receptors. Stimulation by ICI 182 780, G1 and E2-BSA suggests the E2-stimulated IGF-I mRNA expression in BSC cultures is mediated through the GPR30 receptor.     

壳聚糖对仔猪生长性能及血清中类胰岛素生长因子-Ⅰ和生长激素水平的影响

本文旨在研究日粮中添加不同剂量壳聚糖对仔猪生产性能及血清中类胰岛素生长因子-I(IGF-I)和生长激素(GH)水平的影响。试验选择60日龄大白×长白×VI系杂交仔猪96头,随机分为4个日粮处理组,每个处理3个重复,每个重复8头,进行为期21 d的饲养试验。4种日粮分别是在基础日粮中添加0、0 .01 %、0 .05 %、0.1 %的壳聚糖配制而成。结果表明,壳聚糖对仔猪生长性能的影响呈二次剂量依赖关系,添加0 .01 %~0 .05 %壳聚糖能够提高仔猪平均日增重、平均日采食量及增重耗料比,并可提高仔猪血清中IGF-Ι和GH含量。而添加更高水平时生长性能却有下降趋势。因此,在本试验条件下初步得出,0 .01 %~0 .05 %为壳聚糖在仔猪日粮中的适宜添加量。     

IGF-I,GHR and UCP mRNA expression in the liver and muscle of high- and low-feed-efficiency laying Japanese quail at different environmental temperatures

In this study, we analyzed insulin-like growth factor I (IGF-I), growth hormone receptor (GHR) and uncoupling protein (UCP) mRNA expression in the muscle and liver of high- (0.23 g/g) and low- (0.17 g/g) feed-efficiency (FE) Japanese quail at three different air temperatures: comfortable (25 °C), heat stress (38 °C) for 12 h or cold stress (10 °C) for 12 h. Total RNA was extracted from the liver and breast muscle of each quail, and cDNA was amplified using specific primers for the target genes. Expression was analyzed using quantitative real-time PCR (qRT-PCR). IGF-I mRNA expression was higher in the livers of high-FE quail than in the livers of low-FE quail under both heat and cold stress conditions. High-FE birds also showed higher GHR mRNA expression independent of temperature. UCP mRNA expression in the liver was lower in high-FE birds and higher under heat stress compared with the other conditions. IGF-I mRNA expression was higher in the muscle of high-FE quail under the three conditions tested, and UCP mRNA expression was higher under cold stress. Our results suggest that air temperature affects the expression of genes related to growth and mitochondrial energy production, and quail with different feed efficiencies respond differently to environmental stimuli.     

IGF-I与IGFBP-1基因对京海黄鸡生长性状的遗传效应分析

旨在对IGF-I和IGFBP-1基因部分SNPs与鸡生长性状进行关联分析。试验以京海黄鸡为材料,采用PCR-SSCP方法检测2个候选基因的单核苷酸多态性(SNPs),采用一般线性模型(GLM)分析基因型与生长性状的遗传效应。结果表明,IGF-I基因外显子3序列的60bp处有A→G的点突变,在京海黄鸡中检测到AA、AB、BB 3种基因型,A等位基因频率为0.613,B等位基因的频率为0.387;IGFBP-1基因外显子2序列的21bp处有A→T的点突变,104bp处有T→C的突变,在京海黄鸡中检测到CC、CD和DD 3种基因型,C等位基因频率为0.558,D等位基因频率为0.442。IGF-I基因BB基因型个体4周龄体质量显著高于AA和AB型个体(P<0.05);IGFBP-1基因DD基因型个体8周龄体质量显著高于CC基因型个体(P<0.05),4、12和16周龄体质量差异极显著(P<0.01)。因此,推测这些SNPs对京海黄鸡生长性状具有一定的影响,应用于鸡育种过程中的标记辅助选择可以加快育种进程。     

饲粮中添加DL-肉碱复合物和复合胆汁酸对斑点叉尾鮰生长性能的影响

本试验研究了在斑点叉尾鮰饲粮中添加DL-肉碱复合物和复合胆汁酸对其生长性能的影响.采用单因子三重复试验设计,以135尾平均体重(30.6±0.2)g的斑点叉尾鮰随机分为3组.对照组饲喂基础饲粮,DL-肉碱复合物组饲喂在基础饲粮中分别添加DL-肉碱复合物200 mg/kg的饲粮,DL肉碱复合物-复合胆汁酸组饲喂在基础饲粮中添加DL-肉碱复合物200 mg/kg+复合胆汁酸37.5 mg/kg的饲粮.试验期为8周.结果表明:与对照组相比,在饲粮中添加DL-肉碱复合物能够显著提高饲料效率17.28%(P<0.05),显著降低饲料系数15.43%(P<0.05),提高特定生长率12.50%;补充添加DL-肉碱复合物+复合胆汁酸后特定生长率和饲料转化效率较DL-肉碱复合物单独添加组分别降低4.27%和5.76%,差异不显著.     

The effect of exogenous insulin-like growth factor-I (IGF-I) on the reproductive performance of female rats, and on serum concentrations of endogenous IGF-I and IGF-I binding proteins.

The effect of exogenous IGF-I on the reproductive performance of female rats was examined by infusing either recombinant human IGF-I (400 micrograms/d; n = 19) or vehicle (n = 18) over a four-day period (the time of one reproductive cycle) beginning on the day following estrus. The females were exposed to male rats one day after the infusions had commenced, and were euthanized 15 d later. There was no treatment effect on serum progesterone levels at this time or on the number of fetuses. Furthermore, the number of corpora lutea were not different between the IGF-I and vehicle infused groups (15.8 vs. 14.8; P = 0.09). Total serum IGF-I concentrations, as determined with a polyclonal antiserum based RIA, were increased approximately three-fold in samples obtained 20 hr after commencing the IGF-I infusion. These samples were also analyzed for IGF-I with a monoclonal antibody based RIA previously shown to detect human, but not rat, IGF-I. By subtraction, the concentration of endogenous rat IGF-I was found to be approximately 60% higher in IGF-I-infused rats than in control rats. This increase was likely due to a reduced clearance rate of IGF-I from the circulation, caused by a marked induction of 42-46 kDa and 30-34 kDa IGF-I binding proteins observed in these samples with a ligand blot technique. The binding protein induction indicates that the infused IGF-I was bioactive. This induction may have attenuated the effects of IGF-I on ovarian function.     

Effects of colostrum feeding and dexamethasone treatment on mRNA levels of insulin-like growth factors (IGF)-I and -II, IGF binding proteins-2 and -3, and on receptors for growth hormone, IGF-I, IGF-II, and insulin in the gastrointestinal tract of neonatal calves

The somatotropic axis regulates growth of the gastrointestinal tract (GIT). In addition, colostrum feeding and glucocorticoids affect maturation of the GIT around birth in mammals. We have measured mRNA levels of members of the somatotropic axis to test the hypothesis that colostrum intake and dexamethasone treatment affect respective gene expression in the GIT. Calves were fed either colostrum or an isoenergetic milk-based formula, and in each feeding group, half of the calves were treated with dexamethasone (DEXA; 30 microg/kg body weight per day). Individual parameters of the somatotropic axis differed (P < 0.05) among different GIT sections and formula feeding increased (P < 0.05) mRNA levels of individual parameters at various sites of the GIT. Effects of DEXA on the somatotropic axis in the GIT partly depended on different feeding. In colostrum-fed calves, DEXA decreased (P < 0.05) mRNA levels of IGF-I (esophagus, fundus, duodenum, and ileum), IGF-II (fundus), IGFBP-2 (fundus), IGFBP-3 (fundus), IGF1R (esophagus, ileum, and colon), IGF2R (fundus), GHR (fundus), and InsR (esophagus, fundus), but in formula-fed calves DEXA increased mRNA levels of IGF-I (esophagus, rumen, jejunum, and colon). Furthermore, DEXA increased (P < 0.05) mRNA levels of IGF-II (pylorus), IGFBP-3 (duodenum), IGF2R (pylorus), and GHR (ileum), but decreased mRNA levels of IGFBP-2 (ileum), and IGF1R (fundus). Whereas formula feeding had stimulating effects, effects of DEXA treatment on the gene expression of parameters of the somatotropic axis varied among GIT sites and partly depended on feeding.     

Characterization and endocrine regulation of proliferation and differentiation of primary cultured preadipocytes from gilthead sea bream (Sparus aurata)

A preadipocyte primary cell culture was established to gain knowledge about adipose tissue development in gilthead sea bream (Sparus aurata), one of the most extensively produced marine aquaculture species in the Mediterranean. The preadipocytes obtained from the stromal-vascular cell fraction of adipose tissue proliferated in culture, reaching confluence around day 8. At that time, the addition of an adipogenic medium promoted differentiation of the cells into mature adipocytes, which showed an enlarged cytoplasm filled with lipid droplets. First, cell proliferation and differentiation were analyzed under control and adipogenic conditions during culture development. Next, the effects of insulin, GH, and IGF-I on cell proliferation were evaluated at day 8. All peptides significantly stimulated proliferation of the cells after 48 h of incubation (P < 0.002 for GH and IGF-I and P < 0.05 for insulin), despite no differences were observed between the different doses tested. Subsequently, the effects of insulin and IGF-I maintaining differentiation when added to growth medium were studied at day 11, after 3 d of induction with adipogenic medium. The results showed that IGF-I is more potent than insulin enhancing differentiation (P < 0.01 for IGF-I compared with the control). In summary, a primary culture of gilthead sea bream preadipocytes has been characterized and the effects of several regulators of growth and development have been evaluated. This cellular system can be a good model to study the process of adipogenesis in fish, which may help improve the quality of the product in aquaculture.