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        Chicken Insulin-Like Growth Factor-I Stimulates Protein Synthesis of Chicken Embryo Myoblasts Cultured in Serum-Free Medium

        Kita, K.,Okumura, J. Asian Australasian Association of Animal Productio 2001 Animal Bioscience Vol.14 No.1

        The effect of chicken IGF-I on protein synthesis of chicken embryo myoblasts cultured in serum-free medium was examined. When myoblasts were expanded to approximate 20-30% of well, the medium was changed to the serum-free medium including 0, 2, 20, 200 or 2000 ng/ml of recombinant chicken IGF-I. The culture medium including 10% fetal calf serum (FCS) was used as positive control. After 1 day of incubation, protein synthesis was measured by the incorporation of [$^3H$]-L-leucine. Thereafter cells were continued to incubate for further 18 hours, and the radioactivity in the protein was measured as an index of protein synthesis. The values for protein synthesis cultured in the serum-free medium without chicken IGF-I or with 2000 ng/ml of chicken IGF-I were the lowest. Protein synthesis was elevated with increasing chicken IGF-I concentration from 0 to 20 ng/ml. The values for protein synthesis in the 20 ng/ml and 200 ng/ml IGF-I groups were about half of that of the FCS group. The present study revealed that the potency of chicken IGF-I at the levels of 20 to 200 ng/ml to stimulate myoblast protein synthesis was about half of that of 10% FCS.

      • SCIESCOPUSKCI등재

        Effects of Refeeding with a Protein-Free Diets Supplemented with Various Essential Amino Acids on the Plasma Insulin-Like Growth Factor-I Concentration in Fasting Young Chickens

        Kita, K,Shibata, T.,Nagao, K.,Hwangbo, J.,Okumura, J. Asian Australasian Association of Animal Productio 2002 Animal Bioscience Vol.15 No.3

        The effect of refeeding with various single essential amino acids on the recovery of plasma insulin-like growth factor-I (IGF-I) concentration in fasted young chickens was examined. Young chickens (29 days of age) were divided into 15 experimental groups. Chickens in one group were fed on the commercial diet ad libitum for 4 days. The remaining 56 chickens in 14 experimental groups were fasted. After 2 days of fasting, 52 chicks in 13 fasted groups were refed with one of the following experimental diets for 2 days. Eleven experimental diets were protein-free diets supplemented with one of 11 essential amino acids (Arg, Gly, His, Ileu, Leu, Met, Phe, Lys, Thr, Trp, Val). The remaining 2 experimental diets were a protein-free diet containing 11 essential amino acids and a protein-free diet not supplemented with amino acids. Birds in the remaining fasted group continued to be fasted for 2 days. Fasting for 2 days markedly reduced plasma IGF-I concentration. When fasted chickens were refed the protein-free diet containing either Gly alone or all essential amino acids, plasma IGF-I concentration was recovered to the level similar to that of fed chickens. Protein-free diet alone, however, failed to restore the reduced IGF-I concentration in plasma. Body weight loss modulated by feeding with protein-free diets supplemented with various single essential amino acids was associated with changes in plasma IGF-I concentrations. We concluded that body weight loss by feeding with a protein-free diet was lower than that of fasted chickens and that body weight loss associated with the decrease in plasma IGF-I concentration was modulated by feeding with protein-free diets containing various single essential amino acids.

      • SCIESCOPUSKCI등재

        Dietary Fatty Acid Increases Body Weight Gain without a Change in Rumen Fermentation in Fattening Cattle

        Kita, K.,Oka, M.,Yokota, H. Asian Australasian Association of Animal Productio 2003 Animal Bioscience Vol.16 No.1

        Dietary fatty acid including mainly palmitic acid and stearic acid was fed to fattening cattle and its effect on body weight gain, plasma lipid contents and rumen liquid fermentation in vitro was examined. In expt. 1, the effect of dietary fatty acid on body weight gain and plasma lipid concentrations was examined. In the control diet group, cattle were fed 1 kg/day of rice straw and concentrate which satisfied the requirement. In the fatty acid group, cattle were given 250 g/d of fatty acid with the same diet of the control diet group. In the excess concentrate group, cattle were given the same diet of the control diet group plus 735 g/d of concentrate corresponding to the same TDN of 250 g/d of fatty acid. Diets were given for 7 days. Body weight gain of cattle given dietary fatty acid was significantly greater than that of cattle fed only rice straw and concentrate. When dietary fatty acid was added to cattle feed, plasma NEFA and HDL-cholesterol concentrations increased. In expt. 2, the influence of dietary fatty acid on gas production and VFA profile in the rumen liquid was investigated in vitro. In the control group, 10 mg of rice straw and 90 mg of concentrate were incubated in the rumen fluid. In the excess concentrate group, 10 mg of rice straw and 97.5 mg of concentrate were incubated. In the fatty acid group, 10 mg of rice straw, 90 mg of concentrate and 2.5 mg of fatty acids were incubated. The rumen liquid mixed with feed materials was incubated for 24 h and the cumulative gas volume was measured. The VFA profile was also measured. Cumulative gas volume in the rumen liquid with fatty acid was equal to the control. Excess concentrate increased cumulative gas volume compared to the fatty acid group. There was no significant difference in total VFA concentration between experimental diet groups. It is suggested that dietary fatty acid has the potency to improve growth performance in fattening cattle without failure in rumen fermentation.

      • SCIESCOPUSKCI등재

        INFLUENCE OF PHENYLALANINE IN THE MEDIUM ON PROTEIN SYNTHESIS OF CHICKEN EMBRYO FIBROBLASTS

        Kita, K.,Miyazaki, M.,Okumura, J. Asian Australasian Association of Animal Productio 1996 Animal Bioscience Vol.9 No.6

        The influence of phenylalanine (Phe) in the medium on protein synthesis of chicken embryo fibroblasts (CEF) was examined. CEF was derived from 9-d-old embryos by trypsin-EDTA digestion. To examine the deficiency of Phe in the medium, CEF was cultured in Dulbecco's modified Eagle's medium (DMEM) with or without Phe. CEF was also cultured in Dulbecco's phosphate buffered saline (PBS ($Ca^{2+}$, $Mg^{2+}$)) with or without $400{\mu}m$ Phe in order to examine the effect of Phe supplementation. All media were supplemented with 10% (v/v) fetal calf serum. After incubation for 6, 30 and 54 h, protein synthesis was measured by the incorporation of L-[2, $6-^{3}H$] Phe into CEF for further 18 h. Protein synthesis of CEF cultured in DMEM was higher than that in PBS ($Ca^{2+}$, $Mg^{2+}$). High specific radioactivity of Phe due to the low concentration of Phe in the medium resulted in the apparent increase in protein synthesis of CEF. Protein synthesis cultured in PBS ($Ca^{2+}$, $Mg^{2+}$) with Phe did not increase during 72 h of cell culture.

      • SCIESCOPUSKCI등재

        Influence of Chicken Embryo Extract on Protein Synthesis of Chicken Embryo Myoblasts Depends on Cell Density

        Kita, K.,Hiramatsu, K.,Okumura, Jun-ichi Asian Australasian Association of Animal Productio 1998 Animal Bioscience Vol.11 No.6

        The synergistic effect of fetal calf serum (FCS) and chicken embryo extract (CEE) on protein synthesis of chicken embryo myoblasts was examined. Myoblasts were derived from chicken embryo cultured for 14 days by trypsin digestion and cultured in 5% $CO^2/95%$ air at $37^{\circ}C$. When myoblasts were cultured at the low level of cell density (20-50% of well), CEE enhanced the ability of FCS to stimulate protein synthesis of myoblasts. However, there was no significant effect of CEE to stimulate protein synthesis of myoblasts cultured at high level of cell density (100% of well).

      • SCIESCOPUSKCI등재

        Relationship of IGF-I mRNA Levels to Tissue Development in Chicken Embryos of Different Strains

        Kita, K.,Noda, C.,Miki, K.,Kino, K.,Okumura, J. Asian Australasian Association of Animal Productio 2000 Animal Bioscience Vol.13 No.12

        Insulin-like growth factor-I (IGF-I) mRNA levels in the eyes, heart, liver and breast muscle removed from dwarf egg-type, normal egg-type and normal meat-type chicken embryos at 7, 14 and 20 days of incubation were measured. There was no influence of chicken strain on IGF-I gene expression in the eyes and liver. The IGF-I gene expression in eyes increased significantly along with the incubation period. In the liver, IGF-I gene expression at 20 days of incubation was significantly higher than that at 14 days of incubation. In the muscle, the lowest value for IGF-I gene expression was observed in meat-type chicken embryos. Regression analysis revealed that IGF-I gene expression was significantly correlated to the weights of the eyes and liver, but not the muscle. We conclude that there is little influence of strain on tissue IGF-I gene expression in chicken embryos during incubation but that tissue development in chicken embryos is nevertheless at least partly regulated by the change in IGF-I gene expression.

      • SCIESCOPUSKCI등재

        Influence of Refeeding of Protein, Carbohydrate and Fat on Hepatic Insulin-Like Growth Factor-I mRNA Level in Fasted Chicks

        Kita, K.,Hangsanet, K.,Okumura, J. Asian Australasian Association of Animal Productio 1998 Animal Bioscience Vol.11 No.3

        The influence of refeeding either protein, carbohydrate or fat on hepatic insulin-like growth factor-I (IGF-I) mRNA level in chicks which had been fasted for 2 days was examined. The hepatic IGF-I mRNA was measured by ribonuclease protection assay. Fasting reduced hepatic IGF-I mRNA levels to less than half of those in the fed control. When chicks were refed either a control, protein or carbohydrate diet, IGF-I mRNA levels significantly increased to those in the fed control until 2 hours of refeeding. Refeeding of fat did not alter hepatic IGF-I mRNA levels. The significant correlation between liver weight and hepatic IGF-I gene expression suggests that when chicks are refed after 2-d fasting, the acute increase in hepatic IGF-I gene expression brought about after refeeding may be partly regulated by the increase in liver protein metabolism.

      • SCIESCOPUSKCI등재

        MUSCLE PROTEIN SYNTHESIS IN VITRO IN CHICKS FED A LOW-PROTEIN DIET

        Kita, K.,Kuzuya, Y.,Matsunami, S.,Okumura, J. Asian Australasian Association of Animal Productio 1996 Animal Bioscience Vol.9 No.2

        Muscle protein synthesis in vitro was measured in chicks fed low-protein(10% CP) and control(20% CP) diets. Right leg muscles (M. gastrocnemius) were mounted on a support made of stainless steel to stretch in constant tension, whereas left leg muscles were unmounted. Both leg muscles were incubated in Dulbecco's modified Eagle's medium including L-[$4-^3H$] phenylalanine for 60 min to measure in vitro protein synthesis. There was no significant difference in fractional synthesis rate(FSR) of muscle protein between both dietary protein levels, whereas FSR with stretch in constant tension was significantly higher than that without constant tension due to an increase in the absolute synthesis rate(ASR) per unit RNA(the efficiency of RNA to synthesize protein). The ASR of muscle protein in chicks fed the control diet was significantly higher than that in the low-protein diet group.

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