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.     

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.     

Refeeding increases hepatic insulin-like growth factor-I (IGF-I) gene expression and plasma IGF-I concentration in fasted chicks

1. We examined the influence of refeeding after 2 d of fasting on plasma insulin-like growth factor-I (IGF-I) concentration and hepatic IGF-I gene expression in chickens at 6 weeks of age. 2. Hepatic IGF-I mRNA was measured by ribonuclease protection assay and plasma IGF-I concentration was determined by radioimmunoassay. 3. Plasma IGF-I concentration decreased following fasting, increased to the level of fed controls after 2 h of refeeding but then fell back to the level of fasted chickens after 6 h of refeeding. 4. Fasting reduced hepatic IGF-I mRNA concentrations to less than half of those in the fed controls. Refeeding increased IGF-I mRNA sharply at 2 h after refeeding, but by 6 h after refeeding they had taller back again to levels significantly lower than at 2 h. 5. A significant correlation between plasma IGF-I concentration and hepatic IGF-I gene expression was found, suggesting that when chicks are refed after 2 d of fasting, the short-term increase in plasma IGF-I concentration may be partly regulated by the alteration in hepatic IGF-I mRNA.     

The role of endocrine insulin-like growth factor-I (IGF-I) in female bovine reproduction

Insulin-like growth factor-I (IGF-I) plays a pivotal role in cattle fertility, acting as a monitoring signal that allows reproductive events to occur when nutritional conditions for successful reproduction are reached. However, endocrine IGF-I is not a predictor of reproductive events, but rather an indirect estimator of the suitability of the animal to achieve the reproductive event in question. Although measuring circulating IGF-I concentrations might not have any clinical application in the cattle industry, endocrine IGF-I screening will continue to be important for the study of interactions between nutrition and reproduction. In addition, endocrine IGF-I screening could be used as an ancillary test for the selection of cattle for high reproductive potential, especially in herds of high genetic merit for milk production, in which a decline in fertility has been identified.     

Course of epidermal growth factor (EGF) and insulin-like growth factor I (IGF-I) in mammary secretions of the goat during end-pregnancy and early lactation

The epidermal growth factor (EGF) plays a crucial role in mammogenesis in many species. In ruminants, studies are limited, as EGF does not occur in peripheral plasma and specific analytical systems do not exist. Therefore a heterologous radioimmunoassay based on rhEGF was set up to monitor EGF in mammary gland secretions from goats during end-pregnancy and early lactation. IGF-I was measured with an established radioimmunoassay. Samples were collected from 13 goats for 25 days ante-partum and 25 days post-partum. Mammary gland secretions were obtained ante-partum by removing a small amount of the udder secretions (control half) or milking (stimulated half). Post-partum normal milk samples were collected. Blood samples were drawn by jugular venipuncture for the same period. EGF was found to occur in different molecular weight forms in the mammary glands. For routine measurements these proteins were extracted with acetone and not further separated. IGF-I and EGF concentrations in mammary secretions and similarly IGF-I in blood were high ante-partum and decreased slightly towards birth. IGF-I but not EGF is found in the peripheral plasma. Whereas IGF-I concentrations in blood were quite constant post-partum, IGF-I and EGF dropped in mammary secretions close to the detection limits. The decrease was more pronounced in the stimulated half than in the control half. The data support a synergistic role for EGF and IGF-I for mammogenesis. Both factors are further influenced by the milking stimulus and thus the functional state of the udder.     

Evaluation of interference by insulin-like growth factor I (IGF-I) binding proteins in a radioimmunoassay for IGF-I in serum from dairy cows.

Insulin-like growth factor I (IGF-I) circulates in serum bound to a number of different binding proteins (BPs). With antibodies currently available, BPs must be dissociated and inactivated or removed from serum prior to measurement of IGF-I by radioimmunoassay (RIA). Serum samples which spanned a 13-fold range in IGF-I concentration were obtained from lactating dairy cows and used to develop conditions for assay of IGF-I with minimal interference from BPs. Removal of BPs from serum by acid-ethanol extraction resulted in interference in the RIA. Therefore, serum was incubated with 0.1 M glycyl-glycine HCl to inactivate BPs as suggested by Underwood et al. Time, temperature and pH were optimum when serum was incubated for 48 hr at 37 C, pH 3.7. Binding protein inactivation was evaluated by ability of glycyl-glycine incubated serum to reassociate with 125I-IGF-I. In addition, BPs isolated by gel filtration of glycyl-glycine incubated serum were tested for interference in the RIA. The concentration of IGF-I in serum where inactivated BPs were removed by acid gel filtration was compared to corresponding glycyl-glycine incubated serum. There was a 1:1 relationship which intersected at zero indicating that total IGF-I could be measured. Therefore, incubation of serum with glycyl-glycine is a reliable method for measuring total IGF-I in serum from dairy cows.     

Effect of feed restriction on serum somatotropin, insulin-like growth factor-I-(IGF-I) and IGF binding proteins in cyclic heifers actively immunized against growth hormone releasing factor

Feed restriction often increases serum somatotropin (ST) and decreases insulin-like growth factor-I (IGF-I) in ruminants; however, the mechanisms responsible for this change in ST and IGF-I are not well defined. We investigated the effects of feed restriction on serum ST, IGF-I, IGF binding proteins (IGFBP), insulin and nonesterified fatty acids (NEFA) in cyclic Angus and Charolais heifers (n=15) previously immunized against growth hormone releasing factor (GRFi) or human serum albumin (HSAi). Cows were fed a concentrate diet ad libitum (AL) or were restricted to 2 kg cotton seed hulls (R) for 4 d. Each heifer received each dietary treatment in a single reversal design. As anticipated, GRFi decreased ST, IGF-I and insulin (P<.05). In addition, GRFi decreased serum IGFBP-3 (P<.01), but increased IGFBP-2 (P<.01). Feed restriction resulted in an increase in serum ST in HSAi, but not in GRFi heifers. Regardless of immunization treatment, feed restriction decreased serum IGF-I and insulin, and increased NEFA (P<.01). In conclusion, the increase in serum ST levels observed during feed restriction was blocked by active immunization against GRF. However, feed restriction resulted in decreased serum IGF-I in GRFi heifers in spite of initial low levels of IGF-I (due to GRFi). Although GRFi decreased levels of IGFBP-3 and increased levels of IGFBP-2, feed restriction for 4 d did not alter serum IGFBP.     

Recombinant bovine somatotropin (rbST) increases size and proportion of fast-glycolytic muscle fibers in semitendinosus muscle of creep-fed steers

The objective of this study was to determine the effect of recombinant bovine somatotropin (rbST) on muscle fiber histology and histochemistry in creep-fed beef steers. Crossbred steer calves were assigned to one of two treatment groups: control (sham-injected; n = 12) or rbST-injected (0.09 mg x kg(-1) x d(-1); n = 12). Calves were injected every 14 d starting at d 28 of age until weaning at 205 d of age. Biopsies of the semitendinosus muscle were performed on d 100, and slaughter samples of semitendinosus muscle were collected for muscle fiber analyses on d 206. The rbST-treated calves had larger (P = 0.045) fast-twitch-glycolytic (FG) fibers [2,564 +/- 10 vs 2,351 +/- 11 microm2 cross-sectional area, respectively] than controls. No differences (P = 0.36) between rbST-treated and control steers in cross-sectional area were detected for slow-twitch-oxidative (SO) [1,192 +/- 20 vs 1,148 +/- 22 microm2, respectively] or fast-twitch-oxidative-glycolytic (FOG) fibers [1,484 +/- 35 vs 1,403 +/- 38 microm2, respectively]. The percentage distribution for FOG fibers was greater for control calves than for the rbST-treated calves (38.4 vs 34.9 +/- 0.1%, respectively; P = 0.014), whereas the percentage distribution for FG fibers was greater in the rbST-treated calves than for control calves (53.5 vs 48.4 +/- 0.2%, respectively; P = 0.03). The percentage distribution for SO fibers tended to be greater for the control calves than for the rbST-treated calves (13.1 vs 11.7 +/- 0.1%, respectively; P = 0.07). The percentage of FG fibers increased with age (45.4 vs 56.6 +/- 0.8%, respectively; P = 0.001), whereas the percentage distribution of SO (14.3 vs 10.5 +/- 0.5%, respectively) and FOG fibers (40.3 vs 32.9 +/- 0.7%, respectively) decreased (P = 0.001) from d 100 to d 206. The increased longissimus muscle area and dissectable lean tissue in rbST-treated calves are associated with a greater percentage of FG fibers, which possess larger cross-sectional areas than the other fibers.     

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.     

The influence of level of feeding on growth and serum insulin-like growth factor I and insulin-like growth factor-binding proteins in growing beef cattle supplemented with somatotropin

The objective of this study was to determine the effects of level of feeding on growth, feed efficiency (gain:feed; G:F), body composition (BC), and serum concentrations of somatotropin (ST), IGF-I, and IGF-binding proteins (BP) in growing beef cattle supplemented with bovine (b) ST. In each of two consecutive years, 40 growing beef cattle were blocked by weight (average BW: yr 1 = 316 kg, yr 2 = 305 kg) and used in a 2 x 2 factorial arrangement with main effects of bST (0 or 33 microg x kg BW(-1) x d(-1)) and level of feed intake (ad libitum [AL] or 0.75 AL). Relative to uninjected cattle, treatment with bST increased ADG 9.6% (1.14 vs 1.25 kg/d; P < 0.05) and increased G:F 8.1% (12.3 vs 13.3 gain [g]:feed [kg]; P < 0.05), whereas ADG in AL animals was 39% greater than that in 0.75 AL animals (1.39 vs 1.00 kg/d; P < 0.05). There was a tendency (P = 0.10) for a bST x level of feeding interaction, such that the increase in ADG with bST was greater in AL cattle than in 0.75 AL cattle (10.6 vs 7.8%; P = 0.10). Serum concentrations of ST were greater in 0.75 AL cattle than in AL cattle (13.0 vs 8.6 ng/mL; P < 0.05) and in bST-treated cattle than in uninjected cattle (16.3 vs 5.2 ng/mL; P < 0.05). Due to a bST x level of feeding interaction (P < 0.01), the magnitude of the increase in serum ST to exogenous bST was greater (P < 0.01) in 0.75 AL cattle than in AL cattle. Relative to uninjected cattle, treatment with bST increased (P < 0.05) serum concentrations of IGF-I and IGFBP-3 and reduced (P < 0.05) concentrations of IGFBP-2. Similarly, AL cattle had greater (P < 0.05) serum concentrations of IGF-I and IGFBP-3 and reduced (P < 0.05) IGFBP-2 compared with 0.75 AL cattle. In summary, treatment with bST increased growth rate and G:F and stimulated serum IGF-I and IGFBP-3 while reducing IGFBP-2. Feeding at 0.75 ad libitum intake reduced the magnitude of response for each of these variables. Thus, limit-feeding may reduce the effect of exogenous bST on growth rate by blunting bST-induced increases in IGF-I and IGFBP-3 and bST-induced decreases in IGFBP-2.     

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.     

Effect of active immunization against growth hormone releasing factor on concentrations of somatotropin and insulin-like growth factor I in lactating beef cows.

Two experiments were conducted to determine the effects of immunoneutralization of growth hormone-releasing factor [GRF(1-29)-NH2] on concentrations of somatotropin (ST) and insulin-like growth factor I (IGF-I) in lactating beef cows. In Experiment 1, multiparous Hereford cows were immunized against 2 mg GRF(1-29)-(Gly)4-Cys-NH2 conjugated to human serum albumin (GRFi, n = 3) or 2 mg human serum albumin (HSAi, n = 3) at 52 +/- 1 d prior to parturition. Boosters (1 mg) were administered on days 12, 40 and 114 postpartum (pp). Serum samples were collected at 15-min intervals for 5 hr on days 18, 46 and 120 pp, followed by administration (IV) of an opioid agonist (FK33-824; 10 micrograms/kg) and an antagonist (naloxone; .5 mg/kg) at hours 5 and 7, respectively. A GRF-analog ([desamino-Tyr1, D-Ala2, Ala15] GRF (1-29)-NH2; 3.5 micrograms/kg) and arginine (.5 g/kg) were administered at hour 10 on days 47 and 121, respectively. Percentage binding of [125I]GRF (1:100 dilution of serum) 28 d after primary immunization was greater in GRFi (14.3 +/- 4.9) than in HSAi (.7 +/- .3) cows. Binding increased to 29.3 +/- 6.5% after first booster in GRFi cows. Episodic release of ST was abolished by immunization against GRF; concentration and frequency of release of ST were lower (P less than .05) in GRFi than in HSAi cows on all days pp. Concentrations of IGF-I were lower in GRFi than in HSAi cows throughout lactation. Serum ST failed to increase following FK33-824 or arginine in GRFi; however, ST increased after both compounds in HSAi cows. Concentrations of ST following GRF-analog were greater (P less than .05) in HSAi than in GRFi cows. Experiment 2 was conducted to determine if a lower dose of antigen and a single booster would be sufficient to lower ST and IGF-I in lactating cows. Multiparous Hereford and Angus cows were assigned to GRFi (n = 6) or HSAi (n = 6). Primary (1.2 mg) and booster (.5 mg) immunizations were administered -14 and 8 d from calving, respectively. Cows were restricted to 60% of recommended intake of energy during lactation in order to elevate concentrations of ST. Serum samples were collected at 15-min intervals for 6 hr on days 26, 50, 73, 90 and 109 pp. Two of six GRFi cows had binding less than 10% (1:1,000 dilution of serum) and were omitted from further analyses.(ABSTRACT TRUNCATED AT 400 WORDS)     

Moderate doses of porcine somatotropin do not increase plasma insulin-like growth factor-I (IGF-I) or IGF binding protein-3.

The growth rate of the young pig is generally much less than its potential and may be constrained by endocrine status as well as by nutrient intake. The aim of this study was to determine whether porcine somatotropin (pST) could increase growth in the nursing pig. Fourteen sows nursing litters of 6 (n = 7) or 12 (n = 7) piglets were utilized to establish a high and low plane of nutrition for sucking pigs. On Day 4 of lactation, the median two male pigs from each litter were randomly allocated to one of two doses of pST (0 or 60 micrograms/kg/d) until weaning on Day 31. Pigs were bled on Days 4, 13, 22, and 31 of lactation and the plasma was analyzed for insulin-like growth factor (IGF)-I, IGF-II, and IGF binding protein-3 (IGFBP-3). Pigs were weaned into conventional accommodation and further weighed on Days 63, 91, and 119. Pigs from litters of 6 grew more quickly and weighed 2.2 kg (P = 0.01) and 3.5 kg (P = 0.04) more than pigs from litters of 12 at 31 and 63 d of age, respectively. There was no effect of pST on preweaning growth of sucking pigs (261 vs. 258 g/d, P = 0.68), although growth rate increased in the final 3 d before weaning at 31 d (241 vs. 294 g/d, P = 0.01). IGFBP-3 was greater (1.09 vs. 0.78 micrograms/ml, P < 0.001), whereas IGF-I tended to be greater (206 vs. 176 ng/ml, P = 0.14), in pigs from the small litters. There was no effect of pST on plasma IGF-I (182 vs. 195 ng/ml, P = 0.454) or IGFBP-3 (0.93 vs. 0.94 microgram/ml, P = 0.85) concentrations. Plasma IGF-I and IGFBP-3 were highly correlated with the growth rate of nursing pigs (R = 0.638 and 0.756, respectively). There were no effects of pST (340 vs. 328 ng/ml, P = 0.48) or litter size (336 vs. 333 ng/ml, P = 0.88) on IGF-II. In conclusion, pST had no little or no effect on growth performance or plasma IGF-I, IGF-II, or IGFBP-3 in sucking pigs on either a high or low plane of nutrition.     

Temporal pattern of insulin-like growth factor-I response to exogenous bovine somatotropin in lactating cows

The effect of exogenous bovine somatotropin (bST) treatment on the temporal pattern of insulin-like growth factor-I (IGF-I) in serum of four multiparous Holstein cows was examined. Cows (190 +/- 24 days postpartum) were treated with daily subcutaneous injections of recombinant bST (40 mg) or excipient for 12-day periods in a crossover experimental design. During excipient treatment, concentrations of IGF-I in serum were relatively constant throughout the day and averaged 70 ng/ml. Following the first bST injection, serum IGF-I began increasing after a lag of 5 to 7 hr and progressively increased over the first 2 days of treatment. Serum IGF-I levels were approximately 2-fold greater than control values at the end of day 1 of bST treatment, with a 3-fold elevation observed at the end of day 2. Concentrations of IGF-I in serum plateaued by day 3 of bST treatment. Serum concentrations of IGF-I did not follow the oscillating pattern of bST in serum resulting from daily bST injections. Milk yield (3.5% fat-corrected) plateaued after 6 days of bST treatment and was increased 61% (+15.3 kg). Both IGF-I and milk yield remained essentially constant across days for the remainder of treatment. Following cessation of treatment, serum IGF-I and milk yield gradually declined, returning to control values after approximately 4 days. The temporal pattern of circulating concentrations of IGF-I is consistent with a role for IGF-I in mediating a portion of the effects of exogenous bST in lactating cows.     

Localization of insulin-like growth factor I (IGF-I) in the chicken liver after fasting and refeeding: demonstration by using antigen retrieval immunohistochemistry

Immunohistochemical localization of insulin-like growth factor-I (IGF-I) was investigated in the liver of fasted and refed chickens by using an antigen retrieval method. The present study is the first one showing the localization of IGF-I in the chicken liver. Immunoreactivity for IGF-I was detected on the paraffin sections of livers from the fed and refed chickens after the treatment with the antigen retrieval agent. A moderate number of cells showing IGF-I immunoreactivity were scattered in the parenchyma of the liver from fed chickens. These cells were relatively large and polygonal in shape and seemed to be hepatocytes. Reaction products were observed as a granular structure in the cytoplasm of IGF-I-immunoreactive hepatocytes. The number of immunoreactive hepatocytes was increased in the liver from refed chickens compared with fed chickens. Diffuse reaction products as well as granular ones were observed throughout the cytoplasm of IGF-I-immunoreactive hepatocytes of livers from refed chickens. There are, however, no regular patterns of the distribution of immunoreactive hepatocytes in the parenchyma of both fed and refed chickens. In the liver of the fasted chickens, clear immunoreactivity for the peptide was not observed. These data show that IGF-I is located in the chicken hepatocytes and influenced by the nutriture.     

Influence of gonadotropins on insulin- and insulin-like growth factor-I (IGF-I)-induced steroid production by bovine granulosa cells

To determine the effect of gonadotropins on insulin- and insulin-like growth factor (IGF-I)-induced bovine granulosa cell functions, granulosa cells from bovine ovarian follicles were cultured for 2 days in the presence of 10% fetal calf serum (FCS), and then cultured for an additional 2 days in serum-free medium with added hormones. In the presence of 0 or 1 ng/mL of insulin or IGF-I, FSH had little or no effect (P>0.05) on estradiol production by granulosa cells from both small (1–5 mm) and large (≥8 mm) follicles. However, in the presence of ≥3 ng/mL of insulin, FSH increased (P<0.05) estradiol production by granulosa cells from small and large follicles such that the estimated dose (ED₅₀) of insulin necessary to stimulate 50% of the maximum estradiol production was decreased by 2- to 3-fold from 22 to 28 ng/mL in the absence of FSH to 7–14 ng/mL in the presence of FSH. Similarly, in the presence of ≥3 ng/mL of IGF-I, FSH increased (P<0.05) estradiol production by granulosa cells from small and large follicles such that the ED₅₀ of IGF-I for estradiol production was decreased by 4- to 5-fold from 25 to 36 ng/mL in the absence of FSH to 5–6 ng/mL in the presence of FSH. In the presence of FSH, the maximal effect of insulin on estradiol production was much greater than that of IGF-I (137- versus 12-fold increase) and were not additive; when combined, 100 ng/mL of IGF-I completely blocked the stimulatory effect of 100 ng/mL of insulin. In the absence of FSH, the maximal effect of insulin and IGF-I on estradiol production was similar. Concomitant treatment with 30 ng/mL of LH reduced (P<0.05) insulin-stimulated estradiol production by 52% on day 1 and 19% on day 2 of treatment. Insulin, IGF-I and FSH also increased (P<0.05) granulosa cell numbers and progesterone production but their maximal effects were less (i.e., <4-fold increase) than their effects on estradiol production. In conclusion, insulin and IGF-I synergize with FSH to directly regulate ovarian follicular function in cattle, particularly granulosa cell aromatase activity.     

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.     

Effect of feed intake on antimicrobially induced increases in porcine serum insulin-like growth factor I

This study was conducted to determine whether an antimicrobially induced (ASP-250) increase in serum IGF-I was the result of differences in feed intake. Serum IGF-I concentrations were measured in crossbred pigs that were fed a control diet or a diet supplemented with ASP-250 either for ad libitum consumption or limited to 85% of the control pigs' consumption. The pigs that consumed either diet ad libitum, control or ASP-250, consumed similar quantities of feed. The ASP-250 ad libitum-intake pigs had serum IGF-I concentrations that were greater (P<.01) than those of their ad libitum-intake control littermates. Similarly, the ASP-250 limit-fed pigs had serum IGF-I concentrations that were greater (P<.01) than those of the controls. Although the serum IGF-I concentrations of pigs fed the ASP-250-supplemented diet for ad libitum intake were greater than the serum IGF-I concentrations of the pigs limit-fed the ASP-250-supplemented diet, the differences were not significant (P<.08). The ASP-250-fed pigs had higher serum IGF binding protein (BP)-3 concentrations than did their control littermates (P<.003). A time course of antimicrobially induced alterations in serum IGF-I concentrations revealed that the effect of increased serum IGF-I levels in ASP-250-supplemented pigs (P<.02) was observed within 4 d and was maintained throughout the 4-wk study. These findings show that feed intake is not responsible for the increase in serum IGF-I observed with ASP-250 supplementation. Additionally, the antimicrobially induced increase in serum IGF-I concentrations occurs within a few days after initiation of the treatment.     

Origin of plasma insulin-like growth factor-I (IGF-I) during estrus in goats

The objective of this study was to clarify the origin of the increase in plasma insulin-like growth factor-I (IGF-I) during estrus in goats. Focusing on the uterus, the effect of estradiol-17 beta (E2) on the secretion of IGF-I was examined using ovariectomized and hysterectomized animals. A single 5 microg/kg BW of E2 was injected intramuscularly into ovariectomized and hysterectomized goats for 3 consecutive days, and plasma IGF-I concentrations in the two groups were compared. The concentrations of IGF-I rose after the treatments in both groups. The concentrations were significantly higher from 3 to 8 days after the treatment than before the treatment in ovariectomized goats (P<0.05), and from 1 to 3 days after the treatment than before in hysterectomized goats (P<0.05). Thus higher concentrations of plasma IGF-I tended to last longer in ovariectomized than hysterectomized goats. The area under the IGF-I response curve for the 8-day period after the first injection of E2 tended to be greater in ovariectomized than in hysterectomized goats. The results show that E2 increases plasma IGF-I concentrations in goats, and suggest that E2-stimulated IGF-I in plasma may originate mainly from the uterus.