Tatsuo Muramatsu

Energy cost of whole-body protein synthesis measured in vivo in chicks

1. Energy cost of whole-body protein synthesis was measured in vivo in chicks by comparing the changes in protein synthesis and heat production after the administration of cycloheximide, an inhibitor of protein synthesis. 2. Incorporation of phenylalanine into whole-body protein fraction was promptly inhibited after the intravenous injection of cycloheximide, and the effect was sustained for at least 3 hr. 3. Both whole-body protein synthesis and total heat production were significantly reduced by the cycloheximide administration. 4. The energy cost of whole-body protein synthesis was calculated to be 5.35 kJ per g protein synthesis, and hence on a molar basis 7.52 ATPs are required per peptide bond synthesis.

Contribution of whole-body protein synthesis to basal metabolism in layer and broiler chickens.

The effect of starvation on whole-body protein synthesis and on the contribution of protein synthesis to basal metabolic rate was investigated in young chickens (Expt 1). Strain differences between layer and broiler chickens in whole-body protein synthesis and degradation rates were examined when the birds were starved (Expt 2). In Expt 1, 15-d-old White Leghorn male chickens were used, while in Expt 2 Hubbard (broiler) and White Leghorn (layer) male chickens at 14 d of age were used. They were starved for 4 d, and heat production was determined by carcass analysis after 2 and 4 d of starvation. Whole-body protein synthesis rates were measured on 0, 2 and 4 d of starvation (Expt 1), and on 0 and 4 d of starvation (Expt 2). The results showed that starving reduced whole-body protein synthesis in terms of fractional synthesis rate and the amount synthesized. Whole-body protein degradation was increased by starvation both in terms of fractional synthesis rate and the amount degraded on a per kg body-weight basis. Reduced fractional synthesis rate of protein in the whole body was accounted for by reductions in both protein synthesis per unit RNA and RNA:protein ratio. In the fed state, whole-body protein synthesis and degradation rates, whether expressed as fractional rates or amounts per unit body-weight, tended to be higher in layer than in broiler chickens. In the starved state, the difference in the rate of protein synthesis between the two strains virtually disappeared, while the degradation rates were higher in layer than in broiler birds.(ABSTRACT TRUNCATED AT 250 WORDS)

Modification of energy metabolism by the presence of the gut microflora in the chicken

Whether the association with gut microflora modifies the energy metabolism of chickens was investigated by varying the metabolizable energy consumption level from zero to above the maintenance requirement in the germ-free and conventional states. Single comb White Leghorn chicks were either fasted for 3 d (Expt 1), or fed for 6 d at a fixed daily meal intake of 2, 5 or 8 g/d (Expt 2), or 5, 10 or 15 g/d (Expt 3). Changes in carcass energy deposition and heat production indicated that when no dietary energy was available the presence of the gut microflora could benefit the birds by reducing energy losses, whereas when dietary energy was supplied the efficiency of energy utilization was reduced by the presence of the gut microflora. It was concluded, therefore, that the heavy burden of the gut microflora modifies energy metabolism by exerting a buffering or a counter-productive action on the energy utilization of the chicken.

Nutrition and whole-body protein turnover in the chicken in relation to mammalian species

Since the first indication by Schoenheimer (1942) of the dynamic state of body protein, i t is well established that intracellular proteins are subject to substantial synthesis and degradation processes. As a result, accretion of protein in the whole body of animals is the reflection of small differences between synthesis and degradation rates. Because protein accretion, hence overall growth at early stages or egg, wool and milk production at maturity, is one of the major concerns in livestock husbandry as well as for human nutrition, there has been increasing interest in the regulation of these processes in recent years. Investigation into the influence of nutrients on protein synthesis and degradation rates, expressed as either fractional rates (%/d) or absolute rates (g/d or g/kg body~e igh tO”~ per d) in the present review, in the whole body of animals has been greatly facilitated by the establishment of techniques for measuring synthesis rates in vivo such as continuous infusion (Waterlow ef al. 1978) and large dose injection (McNurlan er al. 1979; Garlick et al. 1980b). Currently, estimates of protein degradation still depend mostly on indirect values deduced by subtracting growth rates from synthesis rates of body protein, due primarily to lack of adequate methods. Thus, any speculations relating to protein degradation in the present review are derived from such indirect measurements. Details of problems associated with the measurement of protein synthesis and degradation rates are reviewed elsewhere (Waterlow ef al. 1978; Zak et al. 1979). In the present review, the term ‘protein turnover’ is frequently used. However, its meaning is not always clear to everyone and, as Schimke (1970) pointed out, there has been confusion in the use of the term. In the following discussion, the term ‘protein turnover’

Effect of dietary protein and energy intakes on whole-body protein turnover and its contribution to heat production in chicks

A factorial 3 x 3 experiment was conducted with chicks to investigate the effect of manipulating crude protein (N x 6.25) intake (CPI) and metabolizable energy intake (MEI) simultaneously, in the range low to high (including adequate) levels with regard to the respective requirements, on whole-body protein turnover and its contribution to total heat production. The fractional rate of whole-body protein synthesis was increased curvilinearly by increasing MEI or CPI from low to high levels. In terms of absolute rates whole-body protein synthesis was enhanced by increasing MEI from low to adequate levels, the effect being greater at adequate and high CPI than at low CPI. The effect of varying CPI and MEI on whole-body protein degradation was similar, but less sensitive, to that on whole-body protein synthesis. Increasing MEI from low to high levels elevated total heat production at all CPI levels. There were no interactive effects of varying CPI and MEI on the contribution of whole-body protein synthesis to total heat production, and in general the contribution increased with increasing CPI and decreased with increasing MEI. The contribution of whole-body protein synthesis to total heat production fell within a small range from 11.2 to 16.5%.

Developmental fall in whole body protein turnover of chick embryos during incubation

Whole body protein turnover was measured in chick embryos during incubation to investigate whether or not there is a fall in fractional rates of protein synthesis and degradation during development. Stable isotopically labelled [15N]phenylalanine was injected intraperitoneally into embryos on days 12 and 19. From 60 to 90 min after injection the isotope enrichment in free and protein-bound phenylalanine was measured with a selected-ion gas-chromatograph mass-spectrometer. The results showed that from days 12 to 19 of incubation, there was a remarkable reduction in fractional rates of protein synthesis and degradation in the whole body of chick embryos. During embryonic growth, protein synthesis per unit of RNA that is, the minimum amino acid translation rate of RNA, did not change significantly, whereas the RNA:protein ratio was reduced to one-third from days 12 to 19 of incubation. It was concluded, therefore, that the dramatic fall in fractional synthesis rate in chick embryos would be entirely attributable to the rapid increase in protein content, thereby changing the RNA:protein ratio in parallel with the fractional synthesis rate.

Importance of albumen content in whole‐body protein synthesis of the chicken embryo during incubation

The importance of egg albumen content in whole-body protein synthesis was investigated in developing chicken embryos by using lines genetically selected for high and low albumen contents and by removing albumen from eggs before incubation. 2. Whole-body protein synthesis was estimated by injecting L-[15N]-phenylalanine intravenously on day 12 of incubation. 3. Embryos from high albumen eggs had higher whole-body protein synthesis rates than those from low albumen eggs. 4. Whole-body protein synthesis was reduced by the removal of albumen from eggs before incubation. 5. It was concluded that albumen content per se was of crucial importance in regulating whole-body protein synthesis in chicken embryos during incubation.

Strain differences in whole-body protein turnover in the chicken embryo.

1. Whether or not there is a strain difference in embryonic whole-body protein turnover rates was tested using the chicken embryos of Rhode Island Red carrying a sex-linked dwarf gene (dwarf), White Leghorn (layer), and White Cornish X White Plymouth Rock (broiler) strains on day 12 of incubation. 2. Whole-body protein synthesis was estimated by injecting L-[15N]-phenylalanine either intraperitoneally or intravenously on day 12 of incubation in order to investigate the effect of the route of isotope administration. The results showed that the values for fractional and absolute synthesis rates were approximately 13% higher by intravenous injection than by intraperitoneal injection. 3. Whole-body protein turnover, both in terms of fractional and absolute rates, was significantly faster in dwarf than in broiler embryos, with intermediate values in layer embryos, although no growth differences were observed on day 12. 4. Difference in egg weight, measured before incubation, did not affect protein turnover. 5. It was concluded that the strain difference manifested in whole-body protein turnover of the chicken embryo would probably be a reflection of differences in genetic background.

Gene gun-mediated in vivo analysis of tissue-specific repression of gene transcription driven by the chicken ovalbumin promoter in the liver and oviduct of laying hens.

In order to search tissue-specific elements in the 5′-upstream promoter region, gene gun was used to transfect in vivo plasmid DNAs with varying lengths of truncated ovalbumin promoter fused to the CAT reporter gene to the oviduct and liver of laying hens. The results indicated that in the oviduct, consistently high reporter gene expression was observed irrespective of the length of the truncated ovalbumin gene promoters, whereas in the liver the ovalbumin promoter extending from -3200 to +8 bp suppressed substantially the reporter gene expression compared with consistently high gene expression obtained by the ovalbumin promoters from -2800 to +8 bp or shorter length. It was concluded, therefore, that a tissue-specific silencer-like element might reside most likely in the ovalbumin gene promoter region between -3200 and -2800 bp which represses the ovalbumin gene transcription in the liver, but not in the oviduct of laying hens.

Influence of dietary protein intake on whole-body protein turnover in chicks

1. Experiments were conducted to investigate whether or not varying dietary protein intake affects whole-body protein turnover rates in young chicks. 2. Seven-d-old single comb White Leghorn male chicks were fed on diets with protein concentrations of 0, 100, 200 or 400 g/kg diet under conditions of ad libitum or equalised feeding. At the end of the experiments, the rate of protein synthesis and protein degradation in the whole body were measured in vivo. 3. The results showed that both fractional and absolute rates of protein synthesis increased with increasing dietary protein up to 200 g/kg; above this concentration they remained almost constant when feeding was ad libitum. 4. Similar responses were found with equalized feeding except that a significant reduction in protein synthesis was found when dietary protein was increased from 200 to 400 g/kg diet. 5. Less sensitive and almost parallel changes in protein degradation rates were found. 6. It was concluded that adaptation to varied dietary protein intake occurred primarily through changes in protein synthesis, accompanied by parallel alterations in protein degradation in the whole body.