Effect of photoperiod on hepatic growth hormone receptor 1A expression in steer calves

Photoperiod manipulation, specifically a long-day photoperiod (LDPP), increases milk production in lactating cattle. We have previously reported that the galactopoietic effect of LDPP is associated with an increase in circulating IGF-I, which seems to occur independently of changes in concentrations of GH, IGFBP-2, and IGFBP-3. This study tested the hypothesis that LDPP increases the expression of GH receptor (GHR) 1A messenger RNA (mRNA) in the liver. Two groups of Holstein steer calves (98 +/- 4 d old) were maintained indoors and exposed to LDPP (16-h light: 8-h dark; n = 6) or short-day photoperiod (SDPP; 8-h light: 16-h dark; n = 6) for 60 d. Calves were individually fed a grain- and alfalfa-based diet. Jugular blood samples were collected weekly and via cannula at 15-min intervals for a 4-h period on d 1, 26, and 55 of the study to monitor pulsatile hormone secretion. Serum was harvested and assayed for IGF-I, prolactin (PRL), and GH using RIA. Liver biopsies were obtained at 3-wk intervals to quantify changes in hepatic IGF-I and GHR 1A mRNA using real-time PCR. Steer BW increased during the study but did not differ between treatments. No differences in ADG or total DMI were observed. Relative to SDPP, calves on LDPP had higher (P < 0.05) serum IGF-I concentrations. Concentrations of PRL increased (P < 0.01) in calves exposed to LDPP compared with calves exposed to SDPP. Differences (P < 0.05) in pulsatile GH secretion were also detected. Hepatic IGF-I and GHR 1A mRNA were positively correlated with circulating IGF-I concentrations, and although both increased with time, they were not affected by photoperiod treatment. These results confirm that LDPP increases circulating concentrations of IGF-I, but this occurs independently of changes in IGF-I synthesis and GHR 1A mRNA expression in the liver. Therefore, our hypothesis that LDPP increases the expression of GHR 1A mRNA in the bovine liver is rejected.     

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.     

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.     

Predicting growth in angus bulls: the use of GHRH challenge, insulin-like growth factor-I, and insulin-like growth factor binding proteins

Plasma IGF-I, IGF binding protein-2 (IGFBP-2), and IGFBP-3 were quantified in growing Angus bulls (n = 56) to determine their relationship with postweaning growth and carcass ultrasound measurements. In addition, GH response to GHRH challenge (area-under-the-curve GH [AUC-GH) was determined for each bull as part of a previous study. Blood was collected by jugular venipuncture at the start of a 140-d postweaning growth performance test and at 28 d intervals for plasma IGF-I determination by RIA. Plasma IGFBP-2 and -3 content was measured at the start of the study, on d 70, and d 140 by Western ligand blotting. Individual weights and hip heights were measured every 28 d during the study and carcass longissimus muscle area, intramuscular fat percentage, and carcass backfat were estimated by ultrasound on d 140. Greater plasma IGF-I at the start of the performance test was associated with reduced postweaning ADG and increased longissimus area. Throughout the performance test period, the correlations between plasma IGF-I and hip height were consistently positive, ranging from 0.10 to 0.38, but the correlations between ADG and IGF-I varied from -0.32 to 0.31. Age-adjusted d-1 plasma IGFBP-2 was related to ADG during the performance test, explaining nearly 30% of the variation in ADG. A model combining weaning age, IGFBP-2, and AUC-GH showed a strong relationship with ADG (R2 = 0.40). Plasma IGFBP-2 and -3 were not related to carcass characteristics, and IGFBP-3 was not related to growth rates. This study provides additional evidence for the variable relationship between plasma IGF-I and growth rates in cattle. A significant positive relationship between plasma IGFBP-2, AUC-GH, and postweaning ADG warrants further investigation.     

Effects of L-carnitine on fetal growth and the IGF system in pigs

The effects of L-carnitine on porcine fetal growth traits and the IGF system were determined. Fourth-parity sows were fed a gestation diet with either a 50-g top dress containing 0 (control, n = 6) or 100 mg of L-carnitine (n = 6). At midgestation, fetuses were removed for growth measurements, and porcine embryonic myoblasts (PEM) were isolated from semitendinosus. Real-time quantitative PCR was used to measure growth factor messenger RNA (mRNA) levels in the uterus, placenta, muscle, hepatic tissue, and cultured PEM. A treatment x day interaction (P = 0.02) was observed for maternal circulating total carnitine. Sows fed L-carnitine had a greater (P = 0.01) concentration of total carnitine at d 57 than control sows. Circulating IGF-I was not affected (P = 0.55) by treatment. Supplementing sows with L-carnitine resulted in larger (P = 0.02) litters (15.5 vs. 10.8 fetuses) without affecting litter weight (P = 0.07; 1,449.6 vs. 989.4 g) or individual fetal weight (P = 0.88) compared with controls. No treatment effect was found for muscle IGF-I (P = 0.36), IGF-II (P = 0.51), IGFBP-3 (P = 0.70), or IGFBP-5 (P = 0.51) mRNA abundance. The abundance of IGF-I (P = 0.72), IGF-II (P = 0.34), and IGFBP-3 (P = 0.99) in hepatic tissue was not influenced by treatment. Uterine IGF-I (P = 0.46), IGF-II (P = 0.40), IGFBP-3 (P = 0.29), and IGFBP-5 (P = 0.35) mRNA abundance did not differ between treatments. Placental IGF-I (P = 0.30), IGF-II (P = 0.18), IGFBP-3 (P = 0.94), and IGFBP-5 (P = 0.42) mRNA abundance did not differ between treatments. There was an effect of side of the uterus for IGF-I (P = 0.04) and IGF-II (P = 0.007) mRNA abundance; IGF-I mRNA abundance was greater in the left uterine horn than in the right uterine horn (0.14 and 0.07 relative units, respectively). Placental IGF-II mRNA abundance was greater (P = 0.007) in the left than in the right uterine horn (483.5 and 219.59, respectively). The abundance of IGFBP-3 was not affected by uterine horns in either uterine (P = 0.66) or placental (P = 0.13) tissue. There was no treatment difference for IGF-I (P = 0.31) or IGFBP-5 (P = 0.13) in PEM. The PEM isolated from sows fed L-carnitine had decreased IGF-II (P = 0.02), IGFBP-3 (P = 0.03), and myogenin (P = 0.04; 61, 59, and 67%, respectively) mRNA abundance compared with controls. These data suggest that L-carnitine supplemented to gestating sows altered the IGF system and may affect fetal growth and development.     

Antepartal insulin-like growth factor concentrations indicating differences in the metabolic adaptive capacity of dairy cows

Cows with different Insulin-like Growth Factor-I (IGF-I) concentrations showed comparable expression levels of hepatic growth hormone receptor (GHR). Suppressor of cytokine signaling 2 (SOCS2), could be responsible for additional inhibition of the GHR signal cascade. The aims were to monitor cows with high or low antepartal IGF-I concentrations (IGF-I^high or IGF-I^low), evaluate the interrelationships of endocrine endpoints, and measure hepatic SOCS2 expression. Dairy cows (n = 20) were selected (240 to 254 days after artificial insemination (AI)). Blood samples were drawn daily (day -17 until calving) and IGF-I, GH, insulin, thyroid hormones, estradiol, and progesterone concentrations were measured. Liver biopsies were taken (day 264 ± 1 after AI and postpartum) to measure mRNA expression (IGF-I, IGFBP-2, IGFBP-3, IGFBP-4, acid labile subunit (ALS), SOCS2, deiodinase1, GHR1A). IGF-I concentrations in the two groups were different (p < 0.0001). However, GH concentrations and GHR1A mRNA expression were comparable (p > 0.05). Thyroxine levels and ALS expression were higher in the IGF-I^high cows compared to IGF-I^low cows. Estradiol concentration tended to be greater in the IGF-I^low group (p = 0.06). It was hypothesized that low IGF-I levels are associated with enhanced SOCS2 expression although this could not be decisively confirmed by the present study.     

Follicular Dominance in Cattle Is Associated With Divergent Patterns of Ovarian Gene Expression for Insulin-Like Growth Factor (IGF)-I,IGF-II,and IGF Binding Protein-2 in Dominant and Subordinate Follicles

A decrease in insulin-like growth factor (IGF) binding protein (BP) amount occurs within the follicular fluid of dominant ovarian follicles. At the same time, concentrations of follicular fluid IGF-I do not change. The mRNA for IGF-I, IGF-II, IGFBP-2, and IGFBP-3 in dominant and subordinate follicles were measured to determine if changes in IGF or IGFBP gene expression are associated with follicular dominance. Heifers were ovariectomized during a follicular wave, either during early-dominance (emerging dominant follicle, 9 mm diameter) or mid-dominance (established dominant follicle, 14–16 mm diameter). Follicles were classified as either dominant (DF), subordinate (SF), or not-recruited (NRF; small antral follicles). mRNA was localized by in situ hybridization and measured by image analyses. The IGF-I mRNA (granulosa cells) was greatest in DF and increased in DF, SF, and NRF from early- to mid-dominance. Likewise, IGF-II mRNA (theca cells) was greatest in DF compared with SF or NRF. The IGFBP-2 mRNA (granulosa cells), however, was nearly undetectable in DF, whereas adjacent SF expressed abundant IGFBP-2 mRNA. The NRF were not uniform in their IGFBP-2 expression because only 5 of 13 NRF had IGFBP-2 mRNA. The IGFBP-3 mRNA (granulosa cells) was found only in two NRF, suggesting that local synthesis is not a predominant source of follicular fluid IGFBP-3. These data show that changes in gene expression for IGFBP-2 are opposite to those for IGF-I or IGF-II. Increased IGF-I and IGF-II mRNA and decreased IGFBP-2 mRNA within the DF may be one mechanism leading to follicular dominance. The opposite pattern of IGFBP-2 gene expression in SF and some NRF may lead to follicular atresia.     

Insulin-like growth factor (IGF)-I and IGF binding proteins-2, -3, -4, -5 in porcine corpora lutea during the estrous cycle; evidence for inhibitory actions of IGFBP-3

In this study we measured protein concentrations of insulin-like growth factor (IGF)-I and IGF binding proteins (IGFBPs) 2-5 in porcine corpora lutea (CLs) throughout the estrous cycle (Experiment 1), and examined the effects of IGFBP-3 and IGFBP-3 antibody (AB) on luteal progesterone (P4) secretion in vitro (Experiment 2). For Experiment 1, (CLs) and serum were collected on days (D) 4, 7, 10, 13, 15 and 16 of the estrous cycle (n = 5 animals per day). IGF-I was extracted from CLs and sera, and measured by radioimmunoassay (RIA). IGFBPs were measured in CLs by ligand blots. For Experiment 2, CLs (from Experiment 1) were enzyme dissociated and luteal cells cultured (24 h) in Medium 199 (M199) containing (0-500 ng/ml) IGFBP-3 (+/-IGF-I; 100 ng/ml), or (0-10 microg/ml) IGFBP-3 AB. P4 in media was measured by RIA. In Experiment 1, luteal IGF-I concentrations (ng/g tissue) were maximal on day 4 and gradually decreased thereafter. Serum IGF-I concentrations (ng/ml) were highest on days 4 and 7, compared with days 10-15. Peak levels of luteal IGFBP-3 were also seen on days 4 and 7 of the cycle. Luteal IGFBP-2 concentrations showed a tendency to increase on day 16 (P < 0.05 versus day 10), but no significant changes in IGFBP-4 or -5 were seen. In Experiment 2, IGFBP-3 (w IGF) inhibited the steroidogenic actions of IGF-I, but had no significant actions alone (IGFBP-3 w/o IGF). Finally, IGFBP-3 AB stimulated P4 secretion on days 4 and 7, but not on days 10-16. We conclude that IGFBP-3 inhibits IGF-I actions in the porcine CL.     

Peripheral circulating insulin-like growth factor-I and -II in cattle

Interrelationships between circulating concentrations of the insulin-like growth factors (IGF-I and IGF-II) were investigated in 235 blood samples taken from 145 healthy beef or dairy calves, bulls and cows of different breeds and ages. Autoradiography of Western ligand blots indicated different IGF binding protein (IGFBP) profiles between sera from different categories of cattle. Each IGF radioimmunoassay was validated by determining the effects of IGFBPs, ligand and contraligand, as well as serial dilution and comparison with results obtained after molecular sieve chromatography in acid. In female cattle mean values for IGF-I varied from 5.1 nmol/l in postparturient Holstein cows to 18.5-20.5 nmol/l in growing beef heifers, while mean IGF-II concentrations ranged from 30.0 nmol/l in the cows to 14.7-15.7 nmol/l in the beef heifer calves. In male cattle mean serum IGF-I ranged widely from 8.2 nmol/l in 1-day-old Holstein calves to 67.4 nmol/l in 16-month-old Simmental-type bulls. Mean IGF-II concentrations decreased from 22.9 nmol/l in 1-day-old Holstein bull calves to 11.9 nmol/l in 12-month-old beef bulls. Thus, total molar IGF concentrations were fairly stable in female cattle (24.7-35.1 nmol/l) but extended from 27.3 nmol/l to 81.8 nmol/l in the male cattle. The tendency for a reciprocal relationship between serum concentrations of these growth factors was most obvious in the periparturient cows.     

Reduction in channel catfish hepatic growth hormone receptor expression in response to food deprivation and exogenous cortisol

The objective of this study was to assess the effects of food deprivation and exogenous cortisol administration on somatic growth of channel catfish, Ictalurus punctatus, and examine the resultant changes in circulating insulin-like growth factor-I (IGF-I) concentrations and growth hormone receptor (GHR) gene expression. Integral to this objective, we report the isolation, sequence, and characterization of channel catfish GHR. Sequence analysis and characterization results indicate sequence identity and tissue distribution similar to GHRs in other teleost fish and several functional characteristics conserved in known vertebrate GHRs. The effects of food deprivation and dietary exogenous cortisol administration were assessed as part of a 4-week study. Growth was significantly reduced after 4 weeks in cortisol-fed fish compared to fed-control fish, and fasting resulted in weight loss. At the end of the 4-week study, both IGF-I plasma concentrations and hepatic GHR mRNA abundance were significantly reduced in fasted and cortisol-fed catfish. Levels of hepatic GHR mRNA were positively correlated to circulating IGF-I levels. These results suggest that a reduction in hepatic GHR gene expression might serve as a mechanism for the reduction of circulating IGF-I and growth in channel catfish during periods of food deprivation and stress.     

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.     

A novel phenotype for Lardon dwarfism in miniature Bos indicus cattle suggests that the expression of growth hormone receptor 1A in liver is required for normal growth

Mutations within the growth hormone receptor (GHR) gene that lead to an inactivated or truncated GHR protein cause abnormal growth and small adult size in a variety of species (Laron dwarfism). We studied a line of miniature Bos indicus cattle that have phenotypic (small mature size) and endocrine (increased blood growth hormone and decreased blood insulin-like growth factor-I concentrations) similarities to Laron dwarfs. Liver mRNA from miniature and control cattle was used to amplify a cDNA within the coding region of the GHR. The miniature cattle had GHR mRNA size (determined by Northern blot) and cDNA sequence that were similar to control cattle and, therefore, were unlike most Laron dwarf genotypes in which the GHR gene is mutated. Amounts of mRNA from liver as well as muscle (superficial neck and longissimus) were analyzed by ribonuclease protection assay for IGF-I, total GHR, GHR 1A (inducible, liver-specific GHR mRNA), and GHR 1B (constitutive GHR mRNA). Four control and five miniature bulls were tested. As expected, liver IGF-I mRNA was decreased in the miniature cattle (approximately 12% of control; P < 0.01). The amount of the total GHR as well as GHR 1A mRNA were also decreased in liver (17% and 19% of control, respectively; P < 0.01). Other GHR mRNA, including GHR 1B mRNA, were similar for miniature and control cattle. In muscle, there was a tendency (P < 0.10) for decreased IGF-I mRNA and increased GHR mRNA in miniature compared with control cattle. In summary, a novel phenotype for Laron dwarfism in Bos indicus cattle was associated with underexpression of GHR 1A mRNA, but not other GHR mRNA variants in liver. In addition to decreased GHR 1A mRNA, the miniature cattle had decreased liver IGF-I mRNA. Full expression of GHR 1A in liver, therefore, may be required for full liver IGF-I expression and normal growth.     

Exogenous somatotropin alters IGF axis in porcine endometrium and placenta

The aim of this study was to examine whether exogenous somatotropin (ST) can alter the insulin-like growth factor (IGF) axis in the porcine epitheliochorial placenta. Crossbred gilts were injected either 6 mg of recombinant porcine ST or vehicle from days 10 to 27 after artificial insemination (term day 116). Control and ST-treated gilts were euthanized on day 28 (8 control/5 treated), day 37 (4 control/6 treated), and day 62 (4 control/6 treated) of gestation. Endometrium and placental tissue samples were collected and subjected to mRNA analyses. In control gilts, somatotropin receptor (STR) and IGF-I mRNA abundance in the endometrium decreased with gestation. Conversely, the amounts of IGF-II mRNA and of IGF binding protein (BP)-2 and -3 mRNA, which were analyzed in endometrium and placental chorion, increased with gestation. The endometrium contained less IGF-II mRNA but more IGFBP-2 and-3 mRNA than the placental chorion. In response to pST treatment, the amounts of endometrial STR and IGF-I mRNA were lower at days 28 and 37, but higher at day 62 of gestation. The content of IGF-II mRNA was higher in the endometrium of pST-treated than control gilts on day 37. The amount of IGFBP-2 mRNA was increased on day 37 in endometrium and placenta of pST-treated gilts, whereas no changes in IGFBP-3 mRNA were observed. The IGF-II/IGFBP-2 ratio was higher in the placenta in response to pST on day 28 of gestation. Results show that pST treatment of pregnant gilts during early gestation alters IGF axis in maternal and fetal placental tissues and suggest pST may exert an effect on fetal growth by altering the relative amount of IGFBPs and IGFs at the fetal-maternal interface.     

Insulin administration suppresses an increase in insulin-like growth factor binding protein-2 gene expression stimulated by fasting in the chicken

1. We examined the changes in plasma IGF-I concentration and tissue IGFBP-2 gene expression of young fasted chickens refed a commercial diet or administered bovine insulin intravenously. 2. Plasma IGF-I concentration was decreased by fasting for 2 d. Although plasma IGF-I concentration was increased by refeeding, it didn't recover to the level of chickens fed a commercial diet ad libitum. 3. Insulin administration lowered plasma IGF-I concentration compared to other groups. 4. Hepatic IGFBP-2 mRNA was increased by fasting for 2 d and decreased by refeeding for 6 h. Insulin administration also decreased hepatic IGFBP-2 gene expression stimulated by fasting to the level of refed chickens. 5. IGFBP-2 mRNA in the gizzard was increased by fasting for 2 d and tended to decrease after refeeding for 6 h. Insulin administration decreased gizzard IGFBP-2 gene expression to less than that in refed chickens. 6. There was no between-treatment difference in IGFBP-2 mRNA in the brain and kidney. 7. These results suggest that the changes in IGFBP-2 gene expression in the liver and gizzard by fasting and refeeding might be partly regulated by the alteration in plasma insulin concentration.     

Serum from heifer calves treated with bovine growth hormone affects the rate of proliferation of C2C12 myogenic cells dependent on the plane of nutrition: the role of insulin-like growth factor-I and IGF-binding proteins-2 and -3

The present in vitro experiments were carried out in order to study whether variations in the bovine growth hormone (bGH)/insulin-like growth factor (IGF)-I axis induced by plane of nutrition and bGH treatment of heifer calves caused variations in serum-induced proliferation of C2C12 myoblasts. Serum was obtained from two groups each of six heifer calves (195 +/- 8 kg) before (d -1) and after treatment with 15 mg/day of bGH for 6 days (d 6) fed either a low (GHL) or a high plane (GHH) of nutrition. Preceding the experiment all 12 heifer calves were fed at the low plane of nutrition. At d 6, serum concentrations of insulin and IGF-I were increased while that of IGF-binding proteins (IGFBP)-2 was decreased in GHH, but unchanged in GHL calves. Serum-induced proliferation of C2C12 myoblasts, was elevated at d 6 by GHH treatment. Especially human IGFBP-3 but also bovine IGFBP-2 added to cell cultures inhibited the rate of proliferation of C2C12 myoblasts stimulated by human IGF-I. The present results showed that GH treatment causes changes in the GH/IGF axis, which leads to changes in serum-induced growth of C2C12 muscle cells dependent on the plane of nutrition that mimic in vivo effects of GH treatment, which indicate an endocrine contribution of the IGF system. However, drawbacks of this suggestion are discussed.     

Insulin-like growth factors and IGF-binding proteins in bovine seminal plasma.

Insulin-like growth factor-I (IGF-I) is an important factor for germ cell development and maturation of spermatozoa. Actions of IGFs are modulated by IGF-binding proteins (IGFBPs) that may, depending on their concentration and site of expression, inhibit or enhance effects of IGF-I. We characterized IGFs and IGFBPs in seminal plasma from bulls routinely used for artificial insemination (AI) and from bulls producing poor-quality semen (low mass and individual motility of spermatozoa). IGFs were measured by specific radioimmunoassay in 22 samples of seminal plasma from nine different AI bulls with high (> 76.8%), average (72.8-73.4%), or low (< 69.5%) nonreturn rate (NRR). IGF-I and IGF-II levels were 144 +/- 9 ng/ml (mean +/- SE; range, 79-238 ng/ml) and 144 +/- 10 ng/ml (range, 55-221 ng/ml), respectively, and did not correlate with NRRs. IGF-I concentrations in seminal plasma from bulls producing poor-quality semen (n = 10) were significantly (P < 0.05) greater (194 +/- 26 ng/ml; range, 94-370 ng/ml), whereas IGF-II levels were significantly (P < 0.05) lower (93 +/- 17 ng/ml; range, 38-183 ng/ml) than in AI bulls. Ligand blot analysis of seminal plasma for IGFBPs revealed the presence of a 38-/45-kDa doublet band and a 30-kDa IGFBP. These IGFBPs were identified as IGFBP-3 and IGFBP-5, respectively, by immunoprecipitation using specific antibodies. In addition, a low amount of IGFBP-4 was detected in bovine seminal plasma by immunoprecipitation. There was a marked difference in the activity of IGFBPs between individual bulls, with a relatively small within-bull variance. The differences in IGFBP activities did not correlate with the fertilization capacity of the bulls in vivo or in vitro nor with immunoreactive IGF-I and IGF-II levels in seminal plasma. Our results demonstrate the presence of IGFBPs in bovine seminal plasma. In contrast to human seminal plasma, high activity of IGFBP-3 was detected in seminal plasma of some bulls, suggesting species-specific regulation of IGFBP activity by proteases.     

Feed restriction in young bulls alters the onset of puberty in relationship with plasma insulin-like growth factor-I (IGF-I) and IGF-binding proteins

The objectives of this study were to evaluate the effect of feed restriction and re-alimentation on the onset of puberty and IGF status in peripubertal male calves and to compare the radioimmunoassay (RIA) and western ligand blotting (WLB) methods for bovine IGFBP-2. Twelve prepubertal 290 d-old Belgian Blue bulls (mean weight: +/- 290 kg) were randomly assigned in three groups: a control group (NG; n = 4) receiving a classic fattening diet to induce "normal" growth (1.48 kg/d), a feed restricted group (RG; n = 4) to obtain reduced growth (0.50 kg/d) and, a severely restricted group (SG; n = 4) to nearly stop growth (0.08 kg/d). The feed restriction period was maintained over a period of 114 d. After the period of differential feeding, all animals received the control feed regime over a period of 100 d. Blood samples were collected at fortnightly intervals. Circulating IGF-I was measured by RIA whereas plasma IGFBPs was evaluated by WLB; IGFBP-2 was additionally quantified by RIA procedure. At the beginning of the trial, IGF-I levels were low (<100 ng/ml) and similar in the three groups in accordance with prepubertal status. In the NG group, a progressive rise in IGF-I was observed from Day 42 to Day 142 whereas in the RG and SG groups, IGF-I levels did not change until the experimental restriction period ended. The delay of the rise in plasma IGF-I was longer for the SG group, IGF-I remained low until 2 wk after the end of the period of restricted feeding. Surprisingly, although differences were detected for IGF-I levels between the three groups, the IGFBP-2 and -3 data, evaluated by WLB could only discriminate between NG and SG group and not between NG and RG. However, by using a RIA method, an IGFBP-2 decrease was observed in the NG group coincident with increasing IGF-I levels. For both RG and SG groups, IGFBP-2 levels remained high throughout the feed restriction period whereas plasma IGFBP-2 levels declined upon feeding in both groups. During this feed restriction period, IGFBP-2 was significantly lower in NG than in RG or SG groups. Moreover, SG group animals had higher levels in plasma IGFBP-2 than RG animals. In conclusion, puberty is characterized by developmental changes in plasma IGF-I and IGFBPs that were altered by feed restriction. Moreover, RIA evaluation of plasma IGFBP-2 is able to better reflect group differences than WLB.     

Effects of fasting on IGF-I,IGF-II,and IGF-binding protein mRNA concentrations in channel catfish (Ictalurus punctatus)

The effects of fasting on insulin-like growth factor (IGF)-I, IGF-II, and IGF-binding protein (IGFBPs) mRNA in channel catfish were examined. Fed control fish (Fed) were compared to fish that had been fasted for 30 d followed by 15 d of additional feeding (Restricted). Sequence alignment and similarity to orthologous proteins in other vertebrates provided structural evidence that the 3 catfish sequences identified in the present research were IGFBP-1, -2, and -3. Prolonged fasting (30 d) reduced body weight approximately 60% (P < 0.001) and decreased IGF-I mRNA in the liver and muscle (P < 0.01). Fifteen days of re-feeding restored concentrations of hepatic and muscle IGF-I mRNA. Liver IGF-II mRNA was not affected by fasting but was increased 2.2-fold after 15 d of re-feeding (P < 0.05). Abundance of muscle IGF-II mRNA was similar between the fed control group and the restricted group throughout the experimental period. Fasting also increased liver IGFBP-1 mRNA (P < 0.05) and decreased IGFBP-3 mRNA (P < 0.01), whereas abundance of IGFBP-2 mRNA was not significantly affected. Interestingly, re-feeding for 15 d did not restore concentrations of IGFBP-1 and IGFBP-3 mRNA relative to fed control concentrations. The IGF results suggest that IGF-I and IGF-II are differently regulated by nutritional status and probably have a differential effect in promoting muscle growth during recovery from fasting. Similar to mammals, IGFBP-1 mRNA in catfish is increased during catabolism, whereas IGFBP-3 mRNA is decreased during inhibited somatic growth. The IGFBP results provide additional evidence of the conserved nature of the IGF-IGFBP-growth axis in catfish.