Nobuyoshi Matsunaga

The effect of growth hormone-releasing peptide-2 (KP102) administration on plasma insulin-like growth factor (IGF)-1 and IGF-binding proteins in Holstein steers on different planes of nutrition

This study was conducted to investigate the nutrition-dependent changes in insulin-like growth factor (IGF)-1 and IGF-binding proteins (IGFBPs) with growth hormone releasing peptide-2 (D-Ala-D-betaNal-Ala-Trp-D-Phe-Lys-NH(2); GHRP-2 or KP102) treatment in growing Holstein steers. Eight 13 month-old Holstein steers were grouped on two levels of feed intake (high intake (HI); 2.43% body weight or low intake (LI); 1.22%) and each group was daily injected with KP102 (12.5 microg/kg body weight/day) or saline solution into the jugular vein during 6-day period. The concentration of plasma GH showed an increase after an i.v. bolus injection of KP102 on Day 1 and Day 6 in both the LI and HI groups. Plasma IGF-1 began to increase 10 hr following an i.v. bolus injection of KP102, but this was only observed in the HI group (P < 0.05). Also, the plasma IGF-1 in the HI group with daily injections was significantly greater than the LI group from Day 1 of KP102 administration (P < 0.05). It reached maximum values of 125.1 +/- 7.6 ng/ml after Day 2, and returned to pre-injection levels after Day 4, however, no change in plasma IGF-1 was observed in LI with administration of KP102. During 6 days of treatment, plasma 38-43 kDa IGFBP-3 and 24 kDa IGFBP-4 were significantly higher in KP102 treated steers but only in the HI group (P < 0.05). Plasma 34 kDa IGFBP-2 decreased in the HI group and did not show any change following an injection of KP102. In conclusion, the effect of stimulated endogenous GH with KP102 administration increased plasma IGF-1, 38-43 kDa IGFBP-3 and 24 kDa IGFBP-4 levels in the HI group of growing Holstein steers, but not in the LI one. Thus, we strongly believe that the plasma IGF-1 and IGFBPs response to KP102 treatment is modulated by the nutritional status of growing Holstein steers and the increased plasma IGF-1 concentration with KP102 treatment may be regulated by plasma 38-43 kDa IGFBP-3 and 24 kDa IGFBP-4 in Holstein steers.

Effect of cholinergic blockade on inhibited GH secretion by feeding and intraruminal SCFA infusion in sheep

The effect of cholinergic blockade on suppressed growth hormone (GH) secretion caused by feeding or the intraruminal infusion of an acetate, propionate, and butyrate mixture (107 and 214 mumol.kg-1.min-1 over 6 h) was examined in ovariectomized ewes. Intraruminal infusion at the rate of 107 mumol.kg-1.min-1 increased peripheral plasma short-chain fatty acid (SCFA) concentrations to approximately the physiological levels noted after feeding. Plasma GH was markedly suppressed by feeding and at both the 107 and 214 mumol.kg-1.min-1 SCFA infusion rates; however, cholinergic blocking agents completely blocked the suppressed GH secretion after feeding and only at the 107 mumol.kg-1.min-1 infusion rate. Plasma glucose increased at both infusion rates, and the plasma free fatty acids decreased after feeding and at both infusion rates. However, both metabolites were unchanged relative to the saline control after the injection of the cholinergic antagonists. It is suggested that the decrease in plasma GH observed after feeding and a near-physiological ruminal SCFA increment is mediated via the parasympathetic nerve and not by pharmacological ruminal SCFA increments attributed to other pathways.The effect of cholinergic blockade on suppressed growth hormone (GH) secretion caused by feeding or the intraruminal infusion of an acetate, propionate, and butyrate mixture (107 and 214 μmol ⋅ kg-1 ⋅ min-1over 6 h) was examined in ovariectomized ewes. Intraruminal infusion at the rate of 107 μmol ⋅ kg-1 ⋅ min-1increased peripheral plasma short-chain fatty acid (SCFA) concentrations to approximately the physiological levels noted after feeding. Plasma GH was markedly suppressed by feeding and at both the 107 and 214 μmol ⋅ kg-1 ⋅ min-1SCFA infusion rates; however, cholinergic blocking agents completely blocked the suppressed GH secretion after feeding and only at the 107 μmol ⋅ kg-1 ⋅ min-1infusion rate. Plasma glucose increased at both infusion rates, and the plasma free fatty acids decreased after feeding and at both infusion rates. However, both metabolites were unchanged relative to the saline control after the injection of the cholinergic antagonists. It is suggested that the decrease in plasma GH observed after feeding and a near-physiological ruminal SCFA increment is mediated via the parasympathetic nerve and not by pharmacological ruminal SCFA increments attributed to other pathways.

The effects of growth hormone-releasing peptide-2 (GHRP-2) on the release of growth hormone and growth performance in swine

The effects of GHRP-2 (also named KP102), a new growth hormone-releasing peptide, on the release of growth hormone (GH) and growth performance were examined in swine. The single intravenous (i. v.) injection of GHRP-2 at doses of 2, 10, 30 and 100 microg/kg body weight (BW) to cross-bred castrated male swine stimulated GH release in a dose-dependent manner, with a return to the baseline by 120 min. The peak GH concentrations and GH areas under the response curves (GH AUCs) for 180 min after the injections of GHRP-2 were higher (P < 0.05) than those after the injection of saline. The GH responses to repeated i.v. injections of GHRP-2 (30 microg/kg BW) at 2-h intervals for 6 h were decreased after each injection. The chronic subcutaneous (s.c.) administration of GHRP-2 (30 microg/kg BW) once daily for 30 days consistently stimulated GH release. The GH AUCs for 300 min after the injections on d 1, 10 and 30 of treatment in GHRP-2-treated swine were higher than those in saline-treated swine. However, chronic administration of GHRP-2 caused a partial attenuation of GH response between d 1 and 10 of treatment. The chronic s.c. administration of GHRP-2 also increased average daily gain for the entire treatment period by 22.35% (P < 0.05) and feed efficiency (feed/gain) by 20.64% (P < 0.01) over the saline control values, but did not significantly affect daily feed intake. These results indicate that GHRP-2 stimulates GH release and enhancing growth performance in swine.

Effects of the administration of growth hormone-releasing peptide-2 (GHRP-2) orally by gavage and in feed on growth hormone release in swine

The experiments were conducted to determine the effects of the administration of growth hormone-releasing peptide-2 (GHRP-2, also named KP102), both orally by gavage and in feed, on the release of growth hormone (GH) in swine and to investigate whether attenuation of the GH response occurs after short-term treatment with the peptide in feed. In the first experiment, saline or GHRP-2 at doses of 1, 4.5 and 9 mg/kg body weight (BW) was dissolved in 15 ml saline and administered orally as a bolus by gavage to cross-bred castrated male swine (n = 6). Orally administered GHRP-2 stimulated dose-related increases in peak concentrations of GH, with a return to basal by 120 min. After administering GHRP-2 orally, peak concentrations of GH and areas under the GH response curves (GH AUCs) for 180 min were higher (P < 0.05) than those in saline controls. In Experiment 2, GHRP-2 at doses of 0 (served as control), 1, 4.5 and 9 mg/kg BW was mixed in 150 g of feed and offered to cross-bred castrated male swine (n = 6) at 0900 hr and 1700 hr daily for a 3-d period. Administration of 1 mg/kg BW GHRP-2 to swine in feed failed to stimulate the release of GH, but GHRP-2 at doses of 4.5 and 9 mg/kg BW significantly (P < 0.05) increased plasma concentrations of GH after initial and final treatments at 0900 hr on Days 1 and 3 of treatment, respectively. Peak concentrations of GH and GH AUCs for 180 min after the initial and final treatments in the 4.5 and 9 mg/kg BW GHRP-2-treated swine were higher (P < 0.05) than those in controls. After 3 d of treatment with GHRP-2 in feed at doses of 4.5 and 9 mg/kg BW, GH responses to the peptide were maintained. The results of the present study indicate that the administration of GHRP-2 orally by gavage and in feed stimulates the release of GH in swine, and that the GH-releasing effect of the peptide does not become desensitized after short-term administration in feed.

252 THE ROLE OF INSULIN-LIKE GROWTH FACTOR 1 (IGF-1) IN DEVELOPMENT OF AN ESTROGEN-ACTIVE DOMINANT FOLLICLE DURING THE FIRST FOLLICULAR WAVE POSTPARTUM IN DAIRY COWS

Recent studies have shown that IGF-1 is a crucial factor for ovarian follicular development in mammals. In postpartum (pp) dairy cows, plasma IGF-1 and estradiol (E2) levels in ovulatory cows at the first follicular wave pp are higher than in anovulatory cows. However, the plasma IGF-1 profile in an ovulatory or anovulatory dominant follicle (DF), which have different E2 production, at the first follicular wave pp have not yet been elucidated. Thus, we investigated the changing profile of plasma IGF-1 levels during first follicular wave pp. In 22 multiparous Holstein cows, blood samples were obtained 2 times/week from 4 weeks prepartum to 3 weeks pp, and the first follicular wave was monitored by ultrasound 2 times/week from 7 days pp to ovulatory phase. Detailed IGF-1 profiles in blood were determined during DF growth and maturation 4 times/day from 10 days pp to 7 days after the first ovulation in 5 ovulatory cows and to 20 days pp in 4 anovulatory cows; the data were analyzed by repeated measures ANOVA, and Students t-test. There was no interaction between groups and time within the prepartum or the pp period. The ovulatory cows (n = 13/22) with an estrogen-active dominant (EAD: high plasma E2 level with peak) follicle showed higher IGF-1 levels than anovulatory cows (n = 9/22) with an estrogen-inactive dominant (EID: low plasma E2 level without peak) follicle during the prepartum (117 ± 8 vs. 91 ± 5 ng mL-1; P < 0.05) and the pp (91 ± 4 vs. 64 ± 4 ng mL-1; P < 0.001) period. Especially noteworthy, during the first follicular wave pp in ovulatory cows, the plasma IGF-1 levels were maintained at a high level until E2 levels increased, followed by an LH surge. We observed that the EAD follicle in ovulatory cows ovulated. To further examine the IGF-1 system in the intra-follicular environment, we used the EAD and EID follicles from ovaries of dairy cows obtained at a slaughterhouse. The EAD and EID follicles were classified on the basis of follicle diameter and E2 concentrations in follicular fluid (FF). The significant differences of factors between EAD and EID were analyzed by Students t-test. The expression of IGF-1 mRNA was not detected in follicular cells in either EAD and EID, suggesting that IGF-1 in FF is mainly derived from liver. The free IGF-1 levels in FF in EAD (4.8 ± 0.5 ng mL-1) were higher than those in EID (2.7 ± 0.1 ng mL-1; P < 0.05). In addition, the expression of type 1 IGF receptor (IGFR-1) mRNA in EAD was higher than hat in EID (P < 0.0001). From the results of the present study, it is apparent that the EAD follicle during the first follicular wave pp in ovulatory cows sufficiently expressed IGFR-1, and a liver-derived IGF-1 stimulates E2 production in the follicle to ovulate. In conclusion, our data suggest that a high concentration of IGF-1, secreted from the liver, during the peripartum period may be one of important factors for the appearance of an ovulatory follicle during the first follicular wave pp cows.