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.     

Effect of growth hormone administration to mature miniature Brahman cattle treated with or without insulin on circulating concentrations of insulin-like growth factor-I and other metabolic hormones and metabolites

Previously, we determined that a primary cause of proportional stunted growth in a line of Brahman cattle was related to an apparent refractoriness in metabolic response to GH in young animals. The objective of this study was to determine the effect of administration of GH, insulin (INS), and GH plus INS to mature miniature Brahman cows (n = 6; 9.7 ± 2.06 y; 391 ± 48.6 kg) and bulls (n = 8; 9.4 ± 2.00 y; 441 ± 54.0 kg) on circulating concentrations of metabolic hormones and metabolites, primarily IGF-I and IGF-I binding proteins. We hypothesized that IGF-I secretion could be enhanced by concomitant administration of exogenous GH and INS, and neither alone would be effective. Animals were allotted to a modified crossover design that included four treatments: control (CON), GH, INS, and GH + INS. At the start of the study, one-half of the cattle were administered GH (Posilac; 14-d slow release) and the other one-half served as CON for 7 d. Beginning on day 8, and for 7 d, INS (Novolin L) was administered (0.125 IU/kg BW) twice daily (7:00 AM and 7:00 PM) to all animals; hence, the INS and GH + INS treatments. Cattle were rested for 14 d and then were switched to the reciprocal crossover treatments. Blood samples were collected at 12-hour intervals during the study. Compared with CON, GH treatment increased (P < 0.01) mean plasma concentrations of GH (11.1 vs 15.7 ± 0.94 ng/mL), INS (0.48 vs 1.00 ± 0.081 ng/mL), IGF-I (191.3 vs 319.3 ± 29.59 ng/mL), and glucose (73.9 vs 83.4 ± 2.12 mg/dL) but decreased (P < 0.05) plasma urea nitrogen (14.2 vs 11.5 ± 0.75 mg/dL). Compared with INS, GH + INS treatment increased (P < 0.05) mean plasma concentration of INS (0.71 vs 0.96 ± 0.081 ng/mL), IGF-I (228.7 vs 392.3 ± 29.74 ng/mL), and glucose (48.1 vs 66.7 ± 2.12 mg/dL), decreased (P < 0.01) plasma urea nitrogen (13.6 vs 10.4 ± 0.76 mg/dL), and did not affect GH (13.5 vs 12.7 ± 0.95 ng/mL). In the miniature Brahman model, both the GH and GH + INS treatments dramatically increased circulating concentrations of IGF-I in mature cattle, suggesting that this line of Brahman cattle is capable of responding to bioactive GH.     

Effect of dietary supplementation of chitosan and galacto-mannan-oligosaccharide on serum parameters and the insulin-like growth factor-I mRNA expression in early-weaned piglets

The study was to determine effects of dietary supplementation of chitosan (COS) and galacto-mannan-oligosaccharids (GMOS) on some serum biochemical indices, serum growth hormone (GH) and insulin-like growth factor-I (IGF-I) levels, and hepatic and long gissimus muscle IGF-I mRNA expression in early-weaned piglets. Twenty six Duroc × Landrace × Yorkshire piglets at the age of 15 days were used. The piglets had access to creep feed during the suckling. Six piglets were sacrificed for sampling at the beginning of the study. The other 20 piglets were individually housed in metabolic cages and randomly allotted to four corn and soybean meal-based diets including the control group, the antibiotic group with 110 mg lincomycin/kg diet, the COS group containing 0.025% COS, and the GMOS group with 0.20% GMOS, respectively, in a 2-week feeding experiment. Blood urea nitrogen (BUN) level was reduced whereas serum total protein concentration was increased (P < 0.05) in responses to the COS and GMOS supplementation. Dietary supplementation of COS and GMOS also increased (P < 0.05) the serum GH and IGF-I levels along with enhanced hepatic and the muscle IGF-I mRNA abundance. Dietary supplementation of oligosaccharides such as COS and GMOS may improve growth and feed conversion efficiency by increasing plasma GH and IGF-I levels, in the early-weaned piglets.     

Immunohistochemical expression of progesterone receptors, growth hormone and insulin growth factor-I in feline fibroadenomatous change

The immunohistochemical expression, tissue-specific and cell-specific distribution patterns of progesterone receptors (PR), growth hormone (GH) and insulin growth factor-I (IGF-I) have been studied in 22 cases of feline fibroadenomatous change (FFAC). PR and GH were detected in all cases and were distributed homogeneously throughout the lesion, while IGF-I was detected in 77% of the cases at the site of ductal budding. The simultaneous expression of PR, GH and IGF-I was detected in epithelial cells in 14 of 22 cases while PR and GH expression only was detected in epithelial cells in 11 cases. Cases that expressed GH and IGF-I without PR expression in the stroma were the most numerous. Double immunohistochemical staining showed the co-localisation of PR and GH in a subset of ductal epithelial cells located between basal/myoepithelial and luminal cells (probably undifferentiated stem cells). These results suggest that ligand-activated progesterone receptors may induce the local synthesis of GH which in turn may exert its proliferative action directly and also indirectly through the production of other growth factors, such as IGF-I, in an autocrine/paracrine manner.     

Molecular cloning of insulin-like growth factor-I complimentary DNA and promoter region in the domestic duck (Anas platyrhynchos)*

Complementary DNA (cDNA) and the flanking region of insulin‐like growth factor‐I (IGF‐I) of domesticated duck were cloned. The nucleotide sequence analysis of the cDNA showed seven and eight bases different, respectively, from chicken and turkey IGF‐I cDNA within the coding region. The amino acid sequence of prepro IGF‐I differed by one and two amino acids from those observed in chicken and turkey, respectively. However, no amino acid substitution was observed in the mature IGF‐I region. Sequence analysis of the promoter region and exon 1 of the duck IGF‐I revealed a high degree of similarity to that of the chicken IGF‐I gene. These results suggest that the mechanisms which regulate expression of the IGF‐I gene may be widely conserved in avian species.     

Effects of hypothalamic dopamine on growth hormone‐releasing hormone‐induced growth hormone secretion and thyrotropin‐releasing hormone‐induced prolactin secretion in goats

The aim of the present study was to clarify the effects of hypothalamic dopamine (DA) on the secretion of growth hormone (GH) in goats. The GH‐releasing response to an intravenous (i.v.) injection of GH‐releasing hormone (GHRH, 0.25 μg/kg body weight (BW)) was examined after treatments to augment central DA using carbidopa (carbi, 1 mg/kg BW) and L‐dopa (1 mg/kg BW) in male and female goats under a 16‐h photoperiod (16 h light, 8 h dark) condition. GHRH significantly and rapidly stimulated the release of GH after its i.v. administration to goats (P < 0.05). The carbi and L‐dopa treatments completely suppressed GH‐releasing responses to GHRH in both male and female goats (P < 0.05). The prolactin (PRL)‐releasing response to an i.v. injection of thyrotropin‐releasing hormone (TRH, 1 μg/kg BW) was additionally examined in male goats in this study to confirm modifications to central DA concentrations. The treatments with carbi and L‐dopa significantly reduced TRH‐induced PRL release in goats (P < 0.05). These results demonstrated that hypothalamic DA was involved in the regulatory mechanisms of GH, as well as PRL secretion in goats.     

胰岛素样生长因子及其受体mRNAs在绵羊生殖系统中的表达和分布

为检测胰岛素样生长因子（IGFs）及其受体（IGFR）mRNAs在绵羊发情周期早期卵巢、子宫和输卵管中的表达,探讨绵羊胚胎早期发育过程中其发育环境——生殖道中生长因子的表达、分泌及其作用,取绵羊发情周期早期卵巢、子宫和输卵管,经固定、切片、免疫染色,观察IGFs mRNAs的表达和分布情况。同时用RT-PCR技术研究了各组织中IGF-Ⅰ、IGF-Ⅱ、IGF-ⅠR、IGF-ⅡR mRNAs的表达情况。结果表明,IGFs mRNAs在绵羊发情周期早期的卵巢、子宫和输卵管中都有表达,4种因子表达模式相似：在卵巢中,IGFs主要定位于卵泡颗粒细胞,间质细胞亦有少量表达。在输卵管中,上皮细胞免疫染色呈阳性;在子宫中,腺细胞及上皮细胞的阳性信号强于固有层。RT-PCR检测表明IGFs mRNAs在3种组织中均有表达。     

绵羊生长激素(oGH)的分子克隆与序列测定

本文旨在新疆绵羊生长激素(oGH)基因的克隆。从阿勒泰×中国美利奴杂交羊脑垂体细胞中提取总RNA,分离得到具翻译活性的mRNA。用RTPCR方法扩增出编码oGH的基因,长度为671bp。将扩增产物经琼脂糖凝胶电泳后,回收纯化。采用PCR克隆试剂盒将扩增产物连接至pTAdv质粒上,经PstI和XbaI双酶切鉴定正反插入后,筛选正向插入阳性克隆,并进行序列分析。结果表明克隆到的oGH基因序列与国外报道的序列有4个碱基差异,但所推导的氨基酸序列完全一致。oGH基因的开放阅读框架共含654个核苷酸,编码217个氨基酸,其中信号肽26个氨基酸,编码碱基为1～78(78bp);成熟肽191个氨基酸,编码碱基为79～651(573bp);终止密码子TAG(652～654bp)。其蛋白质的氨基酸序列与牛、人和鱼的序列同源性分别为99.08%、26.7%和5.9%。     

日粮精粗比对育肥藏羊瘤胃组织形态及微生物菌群的影响

本试验旨在研究日粮精粗比对育肥藏羊瘤胃组织形态及微生物菌群的影响.选择210只早期断奶藏羔羊,随机分成7组,每组30只,分别对其饲喂精粗比为20:80(A)、30:70(B)、40:60(C)、50:50(D)、60:40(E)、70:30(F)和80:20(G)的基础日粮,预饲期10 d,试验期90 d.试验结束后,...     

The effect of dietary energy source on serum concentration of insulin-like growth factor-I growth hormone,insulin, glucose,and fat metabolites in weanling horses

Feeding diets high in soluble carbohydrates to growing horses has been implicated in the development of orthopedic diseases; as a result, substitution of dietary fat for soluble carbohydrates has received attention. Because IGF-I is integral to growth and cartilage development and because it is influenced by nutrition, we evaluated the effect of dietary fat substitution on metabolic endpoints and circulating GH and IGF-I in growing horses. Twelve Quarter Horse weanlings, four female and eight male, 151 to 226 d old, were blocked by sex and age and assigned to two treatment groups. Group one (CARB; n = six) was fed a concentrate containing 2.21% fat and 33.9% starch; group two (FAT; n = six) was fed a concentrate containing 10.3% fat and 24.0% starch. Both concentrates contained 3.0 Mcal/kg of DE and 16% CP. Brome hay also was fed. Diets were fed at 0800 and 1600 for 60 d. On d 0, 30, and 60, blood samples were obtained via a jugular catheter from 1 h before until 5 h after the morning feeding. Serum was analyzed for glucose, insulin, GH, IGF-I, NEFA, and total cholesterol (CHOL). Neither ADG (0.85 +/- 0.04 and 0.84 +/- 0.04 kg) nor concentrate DMI (4.04 +/- 0.12 and 4.03 +/- 0.12 kg/d) differed between treatments. There were consistent increases in glucose and insulin in response to feeding on d 0, 30, and 60 for both groups. On d 30, the glucose response to feeding was less (P = 0.07) over time in FAT vs. CARB; however, there were no significant treatment x time effects on d 0 or 60. On d 60, the insulin response to feeding was less (P < 0.05) over time in FAT compared with CARB; however, there was no treatment x time effect on d 0 or 30. Serum CHOL concentrations did not differ between groups on d 0. Horses in the FAT group had increased CHOL concentrations on d 30 and 60 compared with CARB (P < 0.01). Although treatment x time interactions were noted for GH on d 30 and 60 (P < 0.05), only transient and inconsistent differences in the secretory profiles between CARB and FAT treatments were evident at those sampling times. Serum NEFA and IGF-I did not differ between treatments on d 0, 30, or 60. These results suggest that dietary energy source, at least at the level used in this study, did not affect foal growth performance or serum IGF-I and NEFA concentrations. Fat substitution increased serum CHOL and variably affected serum GH, glucose, and insulin concentrations in response to feeding.     

Effect of betaine on growth hormone pulsatile secretion and serum metabolites in finishing pigs

An experiment was conducted to investigate the effect of dietary betaine (0%, 0.125%) on growth performance, growth hormone (GH) pulsatile secretion and serum metabolites in crossbred finishing pigs. Three replications of eight pigs (four barrows and four gilts) were used for each treatment, and blood samples for the determination of GH pulsatile secretion were collected every 15 min for 3 h. The results showed that betaine supplementation resulted in 5.45% (p<0.05) increase in average daily gain, whereas average daily feed intake and feed to gain ratio were not affected. Serum basal GH level, mean GH level and GH pulse amplitude were elevated by 41.79% (p<0.01), 48.98% (p<0.01) and 35.05% (p<0.05), respectively, with betaine treatment, but GH pulse frequency and pulse duration remained unchanged (p>0.05). Serum urea nitrogen concentration in pigs fed betaine was 21.58% lower than that of controls (p<0.01), whereas total protein level was significantly increased with betaine supplementation (p<0.05). The study suggests that betaine could promote growth by enhancing GH secretion in finishing pigs.     

Effect of chromium nanocomposite supplementation on growth hormone pulsatile secretion and mRNA expression in finishing pigs

The study was conducted to investigate the effect of chromium nanocomposite (CrNano) on growth hormone (GH) pulsatile secretion and pituitary GH mRNA expression in finishing pigs. Fifty‐four crossbred pigs (65.57 ± 1.05 kg initial weight) were randomly allotted to one of the three treatments, with three replicate pens per treatment and six pigs per pen. Pigs were offered one of the four diets including a control diet or a control diet supplemented with 200 μg/kg chromium from either chromium chloride (CrCl₃) or CrNano for 35 days. During the trial, all pigs were given free access to feed and water. After completion of the feeding trial, blood samples were taken via auriculares at 15 min intervals for 3 h and three pigs from each treatment were slaughtered to collect pituitary to determine GH mRNA level. The results of GH dynamic secretion showed that supplemental CrNano increased the mean level, the lowest value, peak value and peak duration of GH significantly, while there was no significant effect on peak amplitude. Pituitary mRNA expression of GH was improved significantly in pigs fed diet containing Cr from CrNano. These results indicated that CrNano increased GH mRNA expression and secretion in finishing pigs.     

三价铬对肥育猪生长激素分泌及垂体mRNA表达的影响

将96头体质量约65 kg的杜长大三元杂交猪,按体质量相近、公母各半的原则随机分成4组,每组3个重复.试验猪1组设为对照,饲喂基础饲粮,其余3组为试验组,分别饲喂在基础饲粮基础上添加来源于氯化铬、吡啶羧酸铬,纳米铬的含200 μg/kg铬试验饲粮.试验猪充分饲喂,自由饮水,试验为期40 d.饲养试验结束前1 d,从每组中选择6头猪,间隔15 min颈静脉采血1次,连续3 h,制备血清用于生长激素动态分泌模式研究.饲养试验结束后,按体质量相近原则从每组中选择8头猪进行屠宰,测定背膘厚和背最长肌面积;并各采取3头猪的脑垂体,RT-PCR法测定生长激素mRNA水平.结果表明,饲粮中添加200μg/kg三价纳米铬使肥育猪日增重提高了6.31%(P<0.05),料重比降低了4.61%(P<0.05),背膘厚降低了24.32%(P<0.05),背最长肌面积提高了20.22%(P<0.05).生长激素动态模式分析结果显示,纳米铬组试验猪生长激素总体水平、最低值、峰值和峰持续时间分别提高了42.62%(P<0.05)、87.94%(P<0.05)、26.60%(P<0.05)和17.19%(P<0.05),吡啶羧酸铬组试验猪生长激素总体水平、峰值分别提高了36.58%(P<0.05)和27.18%(P<0.05).垂体生长激素基因RT-PCR分析结果显示,纳米铬组试验猪垂体生长激素mRNA水平提高了27.63%(P<0.05).研究结果提示,三价纳米铬可提高肥育猪垂体生长激素mRNA水平,促进机体生长激素分泌,从而促进生长和改善胴体特性.     

Changes in concentrations of plasma metabolites and insulin-like growth factor-1 after feeding in Japanese Black and Japanese Shorthorn cattle

The aim was to characterize the changes of plasma component concentrations after feeding in Japanese Black (B) and Japanese Shorthorn (S) cattle. Eight females, five B and three S, with a mean bodyweight (standard deviation) of 282.0 (20.3) kg were selected and supplied with only water for 28 h after feeding. Blood was taken from the jugular vein at 4‐h intervals for 32 h after feeding. Metabolite (cholesterol, CL; glucose, GLU; nonesterified fatty acid, NEFA; triacylglycerol, TG; total protein, TP) and insulin‐like growth factor‐1 (IGF‐1) concentrations in plasma were measured. In both breeds GLU tended to decrease linearly with a similar slope. Differences between the initial mean NEFA at 4 h after feeding (Start) and that at 24 h after the start (End) were significant in both breeds, but S increased significantly more than B at and after 8 h from the Start. The TG in S tended to change more than in B, and CL and IGF‐1 in both breeds changed continuously at similar levels. The TP in B tended to decrease more slowly and be lower than S. The change in NEFA, especially, suggests that nutritional status sensitively reflects genetic background.     

The effect of lighting conditions on the rhythmicity of growth hormone secretion in Holstein steers

Growth hormone (GH) secretion regularity and the effects of lighting condition and GH‐releasing hormone (GHRH) on GH release were determined in steers. First, steers were kept under 12:12 L : D conditions (light: 06.00–18.00 hours). The animals were then subjected to a 1‐h advancement in lighting on/off conditions (05.00 and 17.00 hours, respectively). Blood was sampled for 24 h at 1‐h interval on the seventh day of each condition. Second, GHRH was injected intravenously (IV) at 12.00 and 00.00 hours under 12:12 L : D and blood was sampled at 15‐min interval for 4‐h (1 h before and 3 h after the injection). Plasma GH concentrations were measured by a radioimmunoassay. Periodicity of GH secretory profile was calculated by power spectrum analysis using the maximum entropy method. Plasma GH concentrations showed a characteristic pattern consisting of four distinct peaks. Mean periodicity of GH secretory profile was 5.7 h, and it was not altered by any change in lighting conditions. IV injection of GHRH increased GH secretion during the day and night. The increase in GH secretory volume after GHRH injection during the night was equal to that during the day. The present results suggest that GH secreted from the anterior pituitary have regularity in steers.     

日粮中添加不同水平紫苏籽对湖羊生长性能、瘤胃发酵及养分表观消化率的影响

本试验旨在研究日粮中添加不同水平紫苏籽(Perilla frutescens seed,PFS)对湖羊生长性能、血清生化指标、养分表观消化率及瘤胃发酵的影响。选取2月龄、平均初始体重为(23.02±1.36) kg的湖羊公羔60只,依据日粮中PFS含量随机分成为:对照组(Control)、5%紫苏籽组(5%PFS)、10%紫苏籽组(10%PFS)和15%紫苏籽组(15%PFS)。预试期14 d,正试期70 d。饲喂试验结束前21 d,从各组随机挑选4只羊进行7 d的消化代谢试验。试验结束后称重,计算平均日增重(ADG),并采集瘤胃液和血液分别用气相色谱法、比色法、全自动生化分析仪和酶联免疫分析法测定瘤胃液挥发性脂肪酸(VFA)、氨态氮(NH₃-N)、血清中总蛋白(TP)、尿素氮(UN)、葡萄糖(Glu)和血清中生长激素(GH)、胰岛素样生长因子-1(IGF-1)等。结果表明,日粮中PFS水平对湖羊生长性能(干物质采食量、平均日增重和料重比)、血清生化指标(TP、UN、Glu、GH和IGF-1)、瘤胃pH和瘤胃液乙酸、丙酸及总挥发性脂肪酸(TVFA)等含量的影响不显著(P>0.05),但随着PFS添加水平的升高,乙酸、丁酸和TVFA的含量呈降低趋势,丙酸呈上升趋势;与对照组相比,15% PFS组中NH₃-N和乙酸/丙酸极显著降低(P<0.01);添加PFS不影响日粮中粗脂肪(EE)和酸性洗涤纤维(ADF)的表观消化率(P>0.05),但干物质(DM)、有机物(OM)和中性洗涤纤维(NDF)的表观消化率随着PFS添加水平升高而下降,且15% PFS 组的DM和OM的表观消化率均极显著低于对照组(P<0.01);与对照组相比,10% PFS组的粗蛋白(CP)表观消化率极显著提高(P<0.01),而15% PFS组的极显著降低(P<0.01)。综上所述,湖羊日粮中添加PFS水平为10%时,饲喂效果较好。     

谷胱甘肽对肉羊生长性能、屠宰性能及肉品质的影响

选择东北细毛羊×德国肉用美利奴的杂交一代肉羊12只,分成对照组、试验Ⅰ组和Ⅱ组,每组4只,研究谷胱甘肽对肉羊生长性能、屠宰性能及肉品质的影响。试验期60 d。结果表明:与对照组相比,谷胱甘肽显著提高了肉羊的日增重(P<0.05),试验Ⅰ组、Ⅱ组分别提高了14.6%和11.4%;降低了肉羊的料重比(P<0.05),试验Ⅰ组、Ⅱ组分别降低了11.0%和8.1%。试验Ⅰ组的净肉率和GR值显著高于对照组(P<0.05),Ⅱ组的宰前活重显著高于对照组(P<0.05);试验Ⅰ组肉的剪切力显著低于对照组(P<0.05);试验Ⅰ组、Ⅱ组的滴水损失显著低于对照组(P<0.05),而熟肉率显著高于对照组(P<0.05),且两试验组间差异不显著(P>0.05);各试验组宰后45 min内肉的pH没有显著差异(P>0.05),但试验Ⅰ组24 h的pH极显著地高于其他2组(P<0.01)。     

Effects of growth hormone secretagogues on the release of adenohypophyseal hormones in young and old healthy dogs

The effects of three growth hormone secretagogues (GHSs), ghrelin, growth hormone-releasing peptide-6 (GHRP-6), and growth hormone-releasing hormone (GHRH), on the release of adenohypophyseal hormones, growth hormone (GH), adrenocorticotropic hormone (ACTH), thyroid-stimulating hormone (TSH), luteinising hormone (LH), prolactin (PRL) and on cortisol were investigated in young and old healthy Beagle dogs. Ghrelin proved to be the most potent GHS in young dogs, whereas in old dogs GHRH administration was associated with the highest plasma GH concentrations. The mean plasma GH response after administration of ghrelin was significantly lower in the old dogs compared with the young dogs. The mean plasma GH concentration after GHRH and GHRP-6 administration was lower in the old dogs compared with the young dogs, but this difference did not reach statistical significance. In both age groups, the GHSs were specific for GH release as they did not cause significant elevations in the plasma concentrations of ACTH, cortisol, TSH, LH, and PRL. It is concluded that in young dogs, ghrelin is a more powerful stimulator of GH release than either GHRH or GHRP-6. Ageing is associated with a decrease in GH-releasing capacity of ghrelin, whereas this decline is considerably lower for GHRH or GHRP-6.