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.     

Growth hormone treatment of breeding bulls used for artificial insemination improves fertilization rates

To evaluate new therapeutical concepts for male subfertility, we tested the effects of exogenous recombinant bovine growth hormone (rbGH) on various endocrine and metabolic parameters both in blood and in seminal plasma of bulls. Sperm quality was assessed morphometrically and by monitoring the number of successful artificial inseminations (AIs) defined as non-return rates (NRR). Aliquots of 450 semen samples were used from each bull and each experimental period (4 wk before, 14 weeks during and 6 wk after treatment). Six out of ten sires (average age 8.4 years) were treated every two weeks with 640-mg depot formulated rbGH (Eli Lilly). Four bulls received vehicle only. Blood plasma bGH, IGF-I, insulin and glucose concentrations were increased with rbGH treatment. In seminal plasma there was no effect of rbGH treatment on fructose and citrate or on testosterone concentrations. With one exception, rbGH-treated bulls had greater IGFBP-3 concentrations in seminal plasma. Motility of spermatozoa after freezing and thawing was increased compared with pretreatment rates. Most interestingly, the number of successful AIs was increased by an average of 6.0% NRR when ejaculates from rbGH-treated bulls were used.     

Passage of exogenous L-thyroxine and triiodothyronine into bovine ejaculate

The metabolic effects of thyroxine (T4) and triiodothyronine (T3) on spermatozoa metabolism and male anatomy have been demonstrated. The metabolic effects of T3 and T4 could affect the physiologic characteristics of the spermatozoa. There are little data on the passage of T4 and T3 into the ejaculate from blood. The passage of exogenous T4 and T3 from the blood into semen was measured after T4 (45 mg) or T3 (37.5 mg) was injected IV into 8 bulls. Blood and electroejaculate were obtained simultaneously at 20, 40, 60, 120, and 180 minutes and 24 hours after bulls were injected to determine T3 and T4 concentrations compared with base-line values. Blood T3 and T4 concentrations were increased (P less than 0.05) at 20 minutes after bulls were injected (1.1 +/- 0.25 to 598 +/- 76.3 ng/ml and from 66 +/- 5 to 1,318 +/- 105 ng/ml, respectively). Seminal concentrations of T4 were unchanged until 120 minutes after bulls were injected, when they increased (P less than 0.05) from less than 1.2 ng/ml to 4.7 +/- 1.9 ng/ml. However, seminal concentrations of T3 were increased (P less than 0.05) from less than 0.1 ng/ml to 0.5 +/- 0.2 ng/ml at 20 minutes and to 12.5 +/- 2.9 ng/ml at 120 minutes after bulls were injected. It was concluded that exogenous thyroid hormones passed into the ejaculate from blood, with T3 passing faster than T4.     

Plasma testosterone and LH responses to LHRH in double-muscled bulls treated with trenbolone acetate and zeranol

Twenty-four double-muscled Belgian White Blue bulls were assigned, according to body weight, to two groups with and without treatment with anabolic agents. Implants containing 140 mg trenbolone and 36 mg zeranol (Forplix) were inserted sc on the upper face of the ear flap. Plasma concentrations of testosterone and luteinizing hormone (LH) were determined at d 0, 30 and 60 of the experimental period. Mean testosterone levels at d 0 for treated and controls were, respectively, 2.1 and 1.7 ng/ml during the 10-h sampling period. At d 30 and 60, testosterone levels were strongly depressed in implanted bulls (.2 ng/ml) as compared with 2.5 and 1.7 ng/ml in control bulls (P less than .001 at d 30 and P less than .01 at d 60). Average plasma LH concentrations were identical in the two groups at d 0 and 60 (1.1 and 1.5 ng/ml, respectively), but showed a slight decrease at d 30 in the treated group (P less than .10). The pulsatile character of LH and testosterone profiles was abolished by the Forplix treatment. Luteinizing hormone-releasing hormone (LHRH) injection at d 0 was followed in both groups by an immediate and sharp increase in plasma LH concentrations. The LH response reached a maximal value between 20 to 40 min postinjection and then declined rapidly. On the contrary, Forplix treatment strongly reduced LH and testosterone responses to LHRH stimulation in treated animals. Average daily gain and feed to gain ratios were 1.087 +/- .127 and 7.52 +/- .32 kg, respectively, for the control bulls and 1.335 +/- .092 and 6.24 +/- .24 kg for the Forplix-treated bulls, thus clearly showing a beneficial effect of Forplix treatment.     

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.     

Effect of active immunization against growth hormone-releasing factor on growth and onset of puberty in beef heifers.

Angus and Charolais heifers (195 +/- 7 kg) were actively immunized against growth hormone-releasing factor (GRF) to evaluate the effect on concentrations of somatotropin (ST), insulin-like growth factor I (IGF-I), insulin (INS), growth, and onset of puberty. Primary immunizations were given at 184 +/- 7 d of age (d 0 of experiment) by injecting (s.c.) 1.5 mg of GRF-(1-29)-Gly-Gly-Cys-NH2 conjugated to 1.5 mg of human serum albumin (GRFi, n = 22) or 1.5 mg of human serum albumin (HSAi, n = 21). Booster immunizations of .5 mg of antigen were given on d 62, 92, 153, and 251. Antibody binding (percentage at 1:2,000 dilution) to [125I]GRF on d 69 was greater (P less than .01) in GRFi (53.7 +/- 4.5) than in HSAi (10.1 +/- .6) heifers. Serum concentration (ng/ml) and frequency (peaks/5 h) of ST release, respectively, on d 78 were lower (P less than .01) in GRFi than in HSAi heifers (3.3 +/- .1 vs 5.6 +/- .2 and .9 +/- .3 vs 2.3 +/- .2). Serum IGF-I (ng/ml) was lower (P less than .01) in GRFi than in HSAi heifers on d 69 (41 +/- 5 vs 112 +/- 4). Serum INS (microU/ml) on d 78 was lower (P less than .05) in GRFi (2.2 +/- .1) than in HSAi (3.8 +/- .2) heifers. Feed intake, ADG, and feed efficiency were lower (P less than .05) in GRFi than in HSAi heifers. Hip height was lower (P less than .01) and fat thickness was greater (P less than .05) in GRFi than in HSAi heifers by d 132 and 167, respectively. Percentage of heifers attaining puberty (progesterone greater than 1 ng/ml for two consecutive weeks) by d 209 and 379 (12.9 and 18.5 mo of age), respectively, was lower (P less than .05) in GRFi (40.9 and 45.5) than in HSAi (81.0 and 100). In conclusion, growing heifers were successively immunized against GRF. Active immunization against GRF resulted in decreased serum concentration of ST, IGF-I, and INS. In addition, GRF immunization led to lowered feed intake, ADG, and feed efficiency, increased fat depth, and delayed onset of puberty in heifers. We propose that ST and IGF-I are important metabolic mediators involved in the initiation of puberty in heifers.     

Quantitation of bovine macrophage colony-stimulating factor in bovine serum by ELISA

We established an enzyme-linked immunosorbent assay (ELISA) system for the quantitation of bovine macrophage colony-stimulating factor (M-CSF) and used it to measure the serum M-CSF levels in bovine fetuses and calves. The average serum M-CSF level was 2.7+/-1.5 ng/ml in 39 calves under 100 days old, and 1.8+/-0.8 ng/ml in 15 cattle between 101 and 418 days old. Fetal sera samples (n = 6) prepared from cattle between 150 and 280 days of gestational age had a higher average level of M-CSF (8.8+/-1.4 ng/ml). Alteration in serum M-CSF levels in each individual calf was also measured. The serum levels of M-CSF in calves at 0-1 day after birth ranged from 0.52 to 7.3 ng/ml. During the period 113-125 days after birth, serum levels were around 1.4+/-0.39 ng/ml. Although serum M-CSF levels generally decreased as the age of calves advanced, differences among individuals, especially among newborn calves, were observed.     

Active immunization of pigs against growth hormone-releasing factor: effect on concentrations of growth hormone and insulin-like growth factor 1

Cyclic gilts (96 +/- 1 kg) were used to determine the effect of active immunization against growth hormone-releasing factor GRF(1-29)-NH2 on concentrations of growth hormone (GH) and insulin-like growth factor 1 (IGF-1). Gilts were immunized against GRF conjugated to human serum albumin (GRF-HSA, n = 5) or HSA alone at 180 d of age (wk 0). Booster doses were administered at wk 9 and 13. Seven days after the second booster (wk 14), blood samples were collected at 15-min intervals for 6 h before feeding and 30, 60, 120, 180 and 240 min after feeding. Eight days after the second booster, all gilts were administered a GRF analog, [desNH2Tyr1,Ala15]-GRF(1-29)-NH2, followed by an opioid agonist, FK33-824. Blood samples were collected at 15-min intervals from -30 to 240 min after injection. Immunization against GRF-HSA resulted in antibody titers, expressed as dilution required to bind 50% of [125I]GRF, ranging from 1:11,000 to 1:60,000 (wk 11 and 14); binding was not detectable or was less than 50% at 1:100 in HSA gilts (P less than .05). Episodic release of GH was abolished by immunization against GRF-HSA (P less than .05). Mean GH was decreased (P less than .07), but basal GH concentrations were not altered (P greater than .15) by immunization against GRF-HSA. Serum concentrations of IGF-1 were similar at wk 0, but concentrations were lower in GRF-HSA than in HSA gilts (P less than .05) at wk 14.(ABSTRACT TRUNCATED AT 250 WORDS)     

Temporal patterns of growth hormone in blood and seminal plasma of mature dairy bulls

This study was conducted to determine if growth hormone (GH) concentration in bovine seminal plasma would be proportional to but less variable than blood plasma GH. The relationship between GH in blood and seminal plasma was also examined critically. Blood samples were collected at 15-min intervals for 5.75 h, while semen was collected at 30-min intervals over the same time period. Average seminal plasma GH concentrations were 3.2 times higher (P less than .05) than blood plasma GH concentrations (40.4 +/- 15.8 ng/ml vs 12.6 +/- 1.2 ng/ml, respectively). The within animal correlation between blood and seminal plasma was consistently low and nonsignificant (P greater than .05). Overall blood plasma GH and seminal plasma GH concentrations were weakly correlated (r = .418; P greater than .05) among bulls. A predictable relationship between blood and seminal plasma GH concentration does not exist under the conditions of this study.     

Insulin binding to bovine mammary membranes: comparison of microsomes versus smooth membranes

The study was undertaken to determine if membrane preparations of bovine mammary tissue bound insulin. If binding occurred, it was also the intent to compare binding kinetics between microsomes and smooth membranes. Insulin binding to bovine mammary membranes attained equilibrium, was saturable and was specific for insulin. Additional studies showed binding to be pH sensitive and maximal at 10 mM calcium. Binding affinity of insulin to microsomes and smooth membranes was similar, with the exception that smooth membranes bound 1.8 times more insulin per unit of membrane protein than microsomes. Two different methods were used to generate data for kinetic analysis of the insulin-receptor interaction in microsomes. Competitive binding assays (.6 ng [125I]insulin plus 0 to 100 ng insulin) indicated the presence of two binding sites with dissociation constants (Kd) of .32 and 15.8 nM. Direct titration of microsomes with [125I]insulin (.02 to 10 ng/ml) revealed two binding sites with Kd of .017 and .31 nM. The affinity of the second binding site measured by the competitive binding assay method (Kd of 15.8 nM) is low and therefore may not be of physiological importance for insulin action. Insulin appears to bind to two high-affinity receptor sites in bovine mammary microsomes with Kds of .017 nM and .32 nM. These findings show that bovine mammary tissue contains receptors for insulin. In addition, isolation of smooth membranes from microsomes enriches the number of insulin receptors per unit of membrane protein without altering their binding characteristics.     

Neonatal hemiorchidectomy of bulls alters plasma growth hormone levels and advances onset of pubertal testosterone secretion

Bovine GH and testosterone profiles were determined in plasma collected at 20 min intervals during 3 hr bleeding periods on day 25 of life and every 15 days thereafter in six intact (I) Holstein bull calves and in six others which had been hemiorchidectomized (HO) at 10 days of age. In I bulls average plasma GH concentrations varied between 7.9 and 14.5 ng/ml (P greater than 0.05) until 130 days of age, after which the GH level gradually rose (P = 0.007) to a maximum of 19.4 ng/ml on day 205 of life. Episodic release of GH was apparent in 55 day-old and older I bulls and in HO bulls of all ages. Plasma GH concentrations in HO bulls were higher than in I bulls 15 and 30 days after surgery (P = 0.07), at which times the levels in HO bulls averaged 19.6 and 22.5 ng/ml and in I bulls 10.3 and 10.2 ng/ml, respectively. Plasma GH in HO bulls again exceeded that of I bulls at ages of 130-190 days (P = 0.04). Plasma testosterone was virtually nondetectable before 130 days of age in I bulls but thereafter exhibited the typical episodic pattern. In HO bulls, plasma testosterone concentrations began to rise 15 to 30 days before those in I bulls, resulting in an age X treatment interaction (P less than 0.0001). Furthermore, average testosterone levels were higher (P = 0.07) in HO than I bulls at 235 and 250 days of age.(ABSTRACT TRUNCATED AT 250 WORDS)     

Insulin and insulin-like growth factor-I stimulate DNA synthesis in bovine mammary tissue in vitro

Effects of insulin and insulin-like growth factor I (IGF-I) on [3H]thymidine incorporation, in vitro, by mammary tissue slices obtained from prepartum and lactating cows were investigated. Both insulin and IGF-I induced up to a 10-fold increase in [3H]thymidine incorporation in the mammary slices cultured in serum-free media. The effect of insulin-stimulated [3H]thymidine incorporation occurred at a threshold of greater than 1.75 pmol/ml and appeared to reach maximum at greater than 8.8 nmol/ml. The response to IGF-I occurred at greater than 6.5 pmol/ml and reached the equivalent of maximal insulin-stimulated incorporation at 39 pmol/ml. No synergistic or additive effects were observed between these two factors. The in vitro response took 3 to 4 d to reach maximum and was inhibited by cytarabine. Mammary tissue obtained from lactating cows incorporated more [3H]thymidine per microgram DNA in response to insulin (175 pmol/ml) than mammary tissue from pregnant cows. Culture of mammary tissue slices with growth hormone, cortisol, prolactin, or triiodothyronine showed no stimulation of [3H]thymidine incorporation over control. Autoradiography of the cultured lactating tissue showed incorporation of [3H]thymidine by 51, 24 and 29% of the ductal epithelial, secretory alveolar epithelial and myoepithelial cells, respectively. All alveolar epithelial cells that incorporated [3H]thymidine contained secretory products. Among nonsecretory cells, 25 and 28% of the fibroblasts and white blood cells, respectively, were labeled. Insulin-like growth factor I, but not bovine somatotropin, stimulated [3H]thymidine uptake into DNA in lactating bovine mammary tissue. Thus, our data support the concept that bovine somatotropin acts through IGF-I to increase DNA synthesis in mammary cells.     

Serum hormone and metabolite concentrations in fasted young bulls and steers.

The effect of dietary energy restriction on serum insulin, insulin-like growth factor I (IGF-I), growth hormone, (GH), cortisol, plasma urea nitrogen (PUN) and nonesterified fatty acid (NEFA) concentrations was examined. Angus bulls and steers (10 mo) were allotted to two groups of 12 animals and assigned a treatment order. In a switchback design, animals in order 1 were fed a high grain diet, then fasted, while order 2 animals were fasted, and then fed. Animals were allowed 60 hr to acclimate between treatments. Serum and plasma were obtained at 20 min intervals and 60 min, respectively, for 6 hr after feeding and for the last 6 hr of a 30 hr fast. Serum was assayed for insulin, IGF-I, GH, and cortisol (total and free). Plasma was assayed for PUN and NEFA. Mean insulin (ng/ml) differed between fed (.95 +/- .08) and fasted (.26 +/- .08) animals (P less than 01). Both mean total and free cortisol (ng/ml) were lower in fed (11.48 +/- .99) (1.06 +/- .12) than in fasted (17.10 +/- .93) (1.62 +/- .12) animals, respectively (P less than .01). Animals in order 1 differed in mean IGF-I (ng/ml) between fed (199.0 +/- 8.0) and fasted (116.5 +/- 7.2) treatments (P less than .01). Mean IGF-I for animals in order 2 was 146.7 +/- 7.2 in fed and 213.9 +/- 7.2 in fasted animals (P less than .01). Mean GH did not differ between treatments. Mean PUN and NEFA were higher in fasted than in fed animals (P less than .01). Except for % free cortisol (P less than .05), the hormones did not differ between bulls and steers.(ABSTRACT TRUNCATED AT 250 WORDS)     

Bovine lactoferrin binds to insulin-like growth factor-binding protein-3

Insulin-like growth factor (IGF)-binding protein-3 (IGFBP-3) has been shown to have IGF independent actions that appear to be mediated by specific IGFBP-3 binding proteins located on cell membranes. We show here using Western ligand blotting, a number of mammary membrane proteins that bind ¹²⁵I-labeled rhIGFBP-3. Immunoprecipitation studies demonstrated that the >70 kDa protein was identified from bovine mammary microsomes as bovine lactoferrin (bLf). In addition to being a secretory protein, Lf is tightly associated with cellular membranes. Labeled rhIGFBP-3 was shown to bind to commercially purchased and processed apo- or holo-human or bLf, but not bovine transferrin (bTf). Binding of [¹²⁵I]rhIGFBP-3 to other positively charged proteins was not detected nor was binding to rhIGFBP-5 or other mammary-secreted IGFBPs observed. Reciprocal specific binding of [¹²⁵I]bLf to rhIGFBP-3 was shown, but [¹²⁵I]bTf did not show binding to rhIGFBP-3. While [¹²⁵I]rhIGF-II does not bind to bLf, unlabeled rhIGF-II was shown to compete with [¹²⁵I]bLf for rhIGFBP-3 binding. More detailed analysis by dot blot showed that Lf competes (ED₅₀=3 μg/ml) or displaces (ED₅₀=1 mg/ml) bound [¹²⁵I]rhIGF-II from dot blotted rhIGFBP-3. In vitro studies with a bovine primary mammary epithelial cell culture showed that all-trans-retinoic acid stimulates the appearance of bovine IGFBP-3 and bLf in the conditioned media and that [¹²⁵I]rhIGFBP-3 could be utilized to detect conditioned media bLf. These findings reveal a novel role for bLf, binding to IGFBP-3 and perhaps disassociating IGFBP-3:IGF when in high concentration.     

Effect of human growth hormone-releasing factor on bone and mineral metabolism in growing pigs

Little information is available on the effects of growth hormone (GH) and growth hormone-releasing factor (GRF and GHRH) treatment on bone metabolism in pigs. Thus, tibial bending moments and ash contents were studied in 12, 6-wk-old pigs weighing 13 +/- .2 kg. Six pigs (GRF group) were injected s.c. twice daily with 75 micrograms GRF (hGRF [1-29] NH2)/kg BW for 52 d and six remained untreated (control group, C). Average daily gain was slightly (5%; P less than .10) increased in treated pigs. At slaughter, plasma measurements related to calcium homeostasis, such as concentrations of Ca, inorganic P, and vitamin D metabolites (25-OH and 1,25-(OH)2 vitamin D3), were not changed by GRF injection. At slaughter, plasma GH levels were 3.3 times greater in treated (11.3 +/- 3 ng/ml) than in untreated pigs (3.4 +/- .5 ng/ml, P less than .02), whereas those of insulin-like growth factor I were increased by approximately 38%. No difference was observed between the two groups at slaughter in tibial weight, density, bending moment, ash relative to bone volume (29 +/- 1 vs 30 +/- 2 g/100 cm3, GRF vs C), total ash content, or ash relative to dry matter in cortical or medullary bone. Our GRF treatment did not affect bone and mineral metabolism in young, growing pigs.     

Serum insulin-like growth factor I profiles in beef heifers with single and twin pregnancies

Serum samples and BW were obtained from 2-yr-old beef heifers, pregnancy with either single (SF, n = 12) or twin (TF, n = 7) fetuses, at 7-d intervals from d 190 of gestation until calving. Serum insulin-like growth factor I (IGF-I) concentrations of SF heifers gradually declined from d 190 (69.9 +/- 1.0 ng/ml) to d 263 (55.6 +/- .8 ng/ml), then exhibited a slight increase by d 277 (63.4 +/- 1.1 ng/ml). Serum IGF-I concentrations of TF heifers essentially paralleled, yet were lower (P less than .05) than, concentrations in SF heifers for all days tested except d 197 and 205. The SF heifers pregnant with heifer fetuses (n = 6) had higher IGF-I concentrations (P less than .1) than heifers pregnant with bull fetuses (n = 6) for all days tested except d 214 and 235. Instantaneous absolute growth rate (IGR) of SF heifers declined from 1.485 kg/d at d 190 to .257 kg/d by d 277. Rate of decline in IGR of TF heifers was much greater (P less than .0001). Correlations between serum IGF-I concentrations and IGR for SF and TF heifers were .79 (P less than .001) and .59 (P less than .05), respectively. These data suggest that number and sex of fetus influence maternal concentrations of IGF-I and that the combined growth rate of the dam and conceptus during gestation is related to serum IGF-I concentration.     

The effects of bovine recombinant growth hormone administration on insulin-like growth factor-I and the haemopoietic system in thoroughbred geldings

The effect of intramuscularly administered recombinant bovine growth hormone (rbGH) on insulin-like growth factor-I (IGF-I) and white and red blood cell indices was studied in Thoroughbred geldings. An insulin-like growth factor binding protein (IGFBP)-blocked radioimmunoassay was modified and validated for the measurement of IGF-I in equine blood plasma. Baseline values of IGF-I and blood indices were determined over a 48 h period and then a single dose of 5 microg/kg, 10 microg/kg or 50 microg/kg of rbGH was administered. Insulin-like growth factor-I levels increased in a dose-dependent manner, with the highest values between 12 h and 24 h. The highest dose (50 microg/kg) yielded the greatest IGF-I response with a 90.2+/-10.8% increase at 24 h. White blood cell count increased following the three doses of rbGH with the highest white blood cell count at 12 h after the 50 microg/kg dose. Haemoglobin was significantly increased at 24 h (P< 0.05), when values following doses of 10 microg/kg and 50 microg/kg were significantly greater than after the vehicle or the dose of 5 microg/kg. Red blood cell count was not affected by any of the rbGH doses. These results indicated that rbGH is biologically active in the horse and that rbGH at a dose rate of 10 microg/kg or more could be used therapeutically.     

Growth hormone-binding protein in the goat: characterization, evolution under exogenous growth hormone treatment, and correlation with liver growth hormone receptor levels

This report describes the identification and characterization of a specific, high-affinity growth hormone-binding protein (GHBP) in lactating goat serum. Serum samples were incubated with [¹²⁵I]human GH as ligand and in the absence or in the presence of bovine GH as competitor. GH-GHBP complex formation was performed by high-performance liquid chromatography, and the radioactivity was recorded on-line with a Berthold LB detector connected to a computer. The results showed that a serum protein was able to bind specifically to human GH and bovine GH but not to ovine prolactin. Scatchard plots indicated an affinity constant of 4.5 × 10⁸ M⁻¹ and a maximum binding capacity of 4.8 × 10⁻¹⁰ mol/l. In addition, we conducted a 4-wk study to determine the effects of recombinant bovine GH administration on milk production in lactating goats. The effects of recombinant bovine GH treatment on milk production and on the regulation of GHBP and hepatic GH receptor levels were studied. As expected, recombinant bovine GH injected daily increased yields of milk, fat, protein (40, 61, and 40%, respectively), and circulating insulin-like growth factor 1 concentrations compared with controls. During the pretreatment and treatment periods, the control goats exhibited a constant amount of GHBP in serum. No consistent effect of GH treatment on GHBP level was observed. The binding of [¹²⁵I]bovine GH to hepatic microsomal membranes of GH-treated goats was significantly decreased compared with that of control goats. After MgCl₂ desaturation of membranes, the results demonstrated that the down-regulation of GH hepatic receptors, observed for the treated goat group, was induced by receptor occupancy without modification of binding affinity. The GH receptor gene expression, analyzed by slot blot and hybridization with an [-³²P]GH receptor cDNA probe, was not modified by the GH treatment. In lactating goats, the galactopoietic effect of exogenous GH involved a hepatic receptor occupancy. The individual concentration of GHBP in serum cannot explain the individual variations of responses to GH treatment in goats.     

Use of the double-isotope single-injection method for estimating renal function in normal cross-bred swine

The double-isotope single-injection method to estimate renal function which utilizes the radiolabeled chemicals [131I]sodium iodohippurate and [125I]sodium iothalamate was evaluated in swine. A total of 46 normal, cross-bred swine were used to determine the applicability of this method for measuring the glomerular filtration rate and effective renal plasma flow. The mean glomerular filtration rate in pigs was determined to be 5.33 +/- 0.82 ml/kg of body weight/minute for [125I]sodium iothalamate with a biological half-life (T 1/2) of 39.18 +/- 7.44 minutes. The mean effective renal plasma flow was determined to be 19.25 +/- 3.12 ml/kg of body weight/minute for [313I]sodium iodohippurate, with a T 1/2 of 18.45 +/- 1.74 minutes. These values are more closely related to the glomerular filtration and effective renal plasma flow values reported for dogs and cats than they are to values reported for man. The method is rapid and reliable; results are available 6 to 8 hours after the experiment. This method is advantageous when information about renal function variables is a prerequisite to pharmacokinetic or toxicologic studies.