Changes in protein during development of Triturus embryos

Development ◽  
1973 ◽  
Vol 30 (3) ◽  
pp. 647-659
Author(s):  
Hiroshi Imoh ◽  
Tsutomu Minamidani
Keyword(s):  
Protein Synthesis ◽  
Protein Content ◽  
Sodium Carbonate ◽  
Dna Content ◽  
Total Protein ◽  
Soluble Protein ◽  
Nuclear Protein ◽  
Polyacrylamide Gel Electrophoresis ◽  
Tail Bud ◽  
Stage Of Development

The present paper reports basic data on DNA content, protein content, and protein synthesis in Triturus pyrrhogaster embryos during development from cleavage to the hatching stage. Except for measurements of DNA and total protein contents, embryos were labeled with sodium carbonate-14C for 10 h and fractionated into embryonic cell components, i.e. cytoplasmic mass, yolk and pigment granules, and nuclei, in a discontinuous density gradient of sucrose. The protein content and the radioactivity incorporated into protein were measured in each fraction. Those fractions combining protein soluble in buffer at pH 8·3 and in 0·25 N-HCl were further studied with polyacrylamide gel electrophoresis. In the newt embryo, four stages of active DNA increase were observed when cultured at constant temperature; they were gastrula, neurula, late tail-bud, and before-hatching stages. Total protein per embryo decreased from 3 to 2 mg during the development studied. The content of cytoplasmic soluble protein per embryo was low and constant throughout development. Synthesis of the fraction was observed at the earliest stage of development studied though the rate was not high and specific activity of the soluble protein increased during development. Qualitative changes in the newly synthesized protein were observed. With the yolk fraction, synthesis of protein, other than from probable contamination with the cytoplasmic fraction, was not detected and a detailed description was omitted. Changes were observed at two stages of development in the synthesis of nuclear protein soluble in buffer at pH 8·3, the first at gastrulation and the second at late tail-bud stage. The change at gastrulation seemed to be the start of syntheses of the nuclear soluble proteins, while quantitative enhancement rather than qualitative change was noticed at late tail-bud stage. Most of the nuclear protein soluble in 0·25 N-HCI was histone. The histone content increased in accordance with increase in the DNA content and the rate of DNA accumulation was accompanied by proportionate incorporation of radioactivity into histone. Among histone fractions, unique behaviour of the very lysine-rich histone was observed. The availability of [14C]sodium carbonate in rough estimations of protein synthesis in embryos and significance of the data obtained have been discussed.

Protein synthesis and degradation in flight muscles of adult crickets (Gryllus bimaculatus)

1995 ◽  
Vol 198 (5) ◽  
pp. 1071-1077 ◽  
Author(s):  
T Gomi ◽  
T Okuda ◽  
S Tanaka
Keyword(s):  
Protein Synthesis ◽  
Protein Content ◽  
Total Protein ◽  
Muscle Protein ◽  
Polyacrylamide Gel Electrophoresis ◽  
Adult Life ◽  
Total Protein Content ◽  
Gryllus Bimaculatus ◽  
Flight Muscles

The development and degeneration of the flight muscles in adult crickets, Gryllus bimaculatus, were studied (1) by determination of the total protein content, (2) by SDS one-dimensional polyacrylamide gel electrophoresis (SDS­PAGE) of muscle protein and (3) by in vitro culturing of the muscle. The total protein content of the dorso-longitudinal muscle (DLM) and metathoracic dorso-ventral muscle (DVM) increased during the early days of adult life in both sexes. This high protein content was maintained for at least a further 10 days in some individuals, while in others it declined to a low level. Mesothoracic DVMs in males also showed an increase in protein content after adult emergence but did not undergo histolysis, whereas those in females showed no significant temporal change in protein content. Removal of hind wings or artificial de-alation was found to be useful in inducing degeneration of DLMs and metathoracic DVMs. This treatment also stimulated ovarian development in females. An analysis by SDS­PAGE provided no evidence for new protein synthesis prior to or during flight muscle degeneration. A high rate of [3H]- or [35S]methionine incorporation was observed in DLMs taken from newly emerged adults, but, in intact crickets, the rate declined rapidly during the first 3 days of adult life, a pattern consistent with that obtained from the measurement of total protein content. Compared with DLMs removed from intact crickets, DLMs taken from de-alated crickets showed reduced rates of protein synthesis during in vitro culturing. This, together with the onset of protein degradation, appears to cause the rapid decrease in total protein content of the muscle in de-alated crickets.

Total protein content and protein synthesis within pre-elongation stage bovine embryos

1998 ◽  
Vol 50 (2) ◽  
pp. 139-145 ◽  
Author(s):  
J.G. Thompson ◽  
A.N.M. Sherman ◽  
N.W. Allen ◽  
L.T. McGowan ◽  
H.R. Tervit
Keyword(s):  
Protein Synthesis ◽  
Protein Content ◽  
Total Protein ◽  
Total Protein Content ◽  
Bovine Embryos

Glucocorticoid-induced skeletal muscle atrophy in vitro is attenuated by mechanical stimulation

1992 ◽  
Vol 262 (6) ◽  
pp. C1471-C1477 ◽  
Author(s):  
J. A. Chromiak ◽  
H. H. Vandenburgh
Keyword(s):  
Skeletal Muscle ◽  
Protein Synthesis ◽  
Protein Content ◽  
Total Protein ◽  
Weight Lifting ◽  
Mechanical Activity ◽  
Synthesis Rate ◽  
Protein Synthesis Rate

Glucocorticoids induce rapid atrophy of fast skeletal myofibers in vivo, and either weight lifting or endurance exercise reduces this atrophy by unknown mechanisms. We examined the effects of the synthetic glucocorticoid dexamethasone (Dex) on protein turnover in tissue-cultured avian fast skeletal myofibers and determined whether repetitive mechanical stretch altered the myofiber response to Dex. In static cultures after 3-5 days, 10(-8) M Dex decreased total protein content 42-74%, total protein synthesis rates 38-56%, mean myofiber diameter 35%, myosin heavy chain (MHC) content 86%, MHC synthesis rate 44%, and fibronectin synthesis rate 29%. Repetitive 10% stretch-relaxations of the cultured myofibers for 60 s every 5 min for 3-4 days prevented 52% of the Dex-induced decrease in protein content, 42% of the decrease in total protein synthesis rate, 77% of the decrease in MHC content, 42% of the decrease in MHC synthesis rate, and 67% of the decrease in fibronectin synthesis rate. This in vitro model system will complement in vivo studies in understanding the mechanism by which mechanical activity and glucocorticoids interact to regulate skeletal muscle growth.

scholarly journals Effects of oestradiol-17β and progesterone on total and nuclear-protein synthesis in epithelial and stromal tissues of the mouse uterus, and of progesterone on the ability of these tissues to bind oestradiol-17β

1970 ◽  
Vol 119 (4) ◽  
pp. 773-784 ◽  
Author(s):  
J. A. Smith ◽  
L. Martin ◽  
R. J. B. King ◽  
M. Vértes
Keyword(s):  
Protein Synthesis ◽  
Total Protein ◽  
Nuclear Protein ◽  
Gradient Centrifugation ◽  
Sucrose Density Gradient ◽  
Nuclear Proteins ◽  
Sucrose Density Gradient Centrifugation ◽  
Mouse Uterus

1. A method is described for separating uterine epithelium that is 80% pure and connective-tissue stroma that is 60% pure. This was used to study the effects of steroid hormones on total and nuclear-protein synthesis in these tissues. 2. Oestradiol-17β given alone produces mitoses in the epithelium but not in the stroma. It stimulated incorporation in vitro of [14C]lysine into total protein, histones and acidic nuclear proteins to a greater extent in epithelium than stroma. Incorporation into acidic nuclear proteins was most markedly stimulated, reaching four to six times the normal value 4h after treatment, and then declining rapidly. This peak was only seen in epithelial preparations. 3. After pretreatment with progesterone, oestradiol-17β has the reverse effect, producing mitoses only in stroma. Progesterone alone had no effect on the amounts or rates of incorporation of [14C]lysine into stromal nuclear proteins, but changes after oestradiol-17β treatment were similar to those seen in epithelium with oestradiol-17β alone. In the epithelium, progesterone alone depressed incorporation into histones and acidic nuclear proteins, but did not abolish the subsequent response to oestradiol-17β. With this treatment there was a rapid, large and transient increase in incorporation into epithelial total protein not seen with oestradiol-17β alone. 4. Progesterone had no qualitative effect on the distribution of specific oestrogen-binding proteins, as judged by sucrose-density-gradient centrifugation. However, progesterone treatment increased the uptake in vivo of [6,7-3H]oestradiol-17β by stroma, and it is possible that this is important although the differences were not apparent after labelling in vitro.

scholarly journals The influence of live weight, live-weight change and diet on protein synthesis in the skin and skeletal muscle in young Merino sheep

1998 ◽  
Vol 79 (3) ◽  
pp. 267-274 ◽  
Author(s):  
S. M. Liu ◽  
G. Mata ◽  
H. O'Donoghue ◽  
D. G. Masters
Keyword(s):  
Protein Synthesis ◽  
Protein Content ◽  
Total Protein ◽  
Live Weight ◽  
Rapeseed Meal ◽  
Major Protein ◽  
Skin Area ◽  
Wool Growth ◽  
Period 2 ◽  
Nutrient Partitioning

Wool growth is derived directly from protein synthesis in the skin of sheep, and is affected by the nutritional status of the animals. The present experiment examined both protein synthesis in the skin and muscle and wool growth in Merino lambs differing in live weight, intake and dietary protein source. The experiment was a 23 factorial design: twenty-four 5-month-old lambs initially weighing 33 kg (heavy) or 25 kg (light) were fed on a hay-based diet with either lupin seed or rapeseed meal as the major protein sources to maintain live weight (M) for 56 d, or were fed at 0.6M for 28 d (period 1) followed by 28 d at 1.6M (period 2). Fractional protein synthesis rates (FSR, % per d) in the skin and the m. longissimus dorsi on days 4 and 24 of period 1 and day 4 of period 2 were measured by means of a flooding dose of l-[ring-d5]phenylalanine, and wool growth on a skin patch over period 1 was also measured. The FSR ranged from 13.2 to 20.2% per d in the skin, higher than reported for other breeds, and 1.53–3.07% per d in the muscle. Sheep on the low intake (0.6M) had significant reductions in FSR, protein content (g), protein synthesis (g/d) in the skin, and wool growth (g/d). The heavy lambs had similar FSR to the light lambs, but had a higher skin protein content and total protein synthesis per unit of skin area (100 cm2) and, therefore, grew more wool. The rapeseed-meal diet increased FSR and wool growth only in the light lambs over the short term. The protein deposited in wool over period 1 was 0.185 of the total protein synthesis in the skin, regardless of live weight, intake or diet, a result similar to other breeds. With the changes in dietary intake, protein synthesis in the skin and muscle responded differentially, with nutrient partitioning at sub-maintenance in favour of wool growth but at supra-maintenance, following a nutrient restriction, in favour of weight gain in young growing sheep.

scholarly journals Corticotropin regulation of the synthesis of a specific rat adrenal cytosolic protein. Effects of hypophysectomy and actinomycin D

1979 ◽  
Vol 182 (3) ◽  
pp. 717-725 ◽  
Author(s):  
Alice Dazord ◽  
Dominique Gallet ◽  
Helene Cohen ◽  
Jose M. Saez
Keyword(s):  
Protein Synthesis ◽  
Cyclic Amp ◽  
Total Protein ◽  
Actinomycin D ◽  
Polyacrylamide Gel Electrophoresis ◽  
Cytosolic Protein ◽  
And Control ◽  
Corticotropin Stimulation ◽  
Stimulation Of

The mechanism of corticotropin stimulation of the synthesis of a specific rat adrenal cytosolic protein was investigated. This protein (protein E) has a mol.wt. of approx. 30000. It is detected by polyacrylamide-gel electrophoresis of cytosol prepared from adrenal slices from rats treated with corticotropin in vivo and control rats, the slices being incubated with [3H]- and [14C]-leucine respectively. In rats 1–15 days after hypophysectomy, corticotropin, like dibutyryl cyclic AMP, induces an increase in protein E similar to that induced in control rats, even though both compounds no longer stimulate total protein synthesis. Corticotropin stimulation of protein E synthesis is mediated by cyclic AMP but not by corticosterone, since aminoglutethimide, a steroidogenic inhibitor, does not affect corticotropin stimulation, and dexamethasone alone has no effect. Actinomycin D, when injected in vivo 1h before or after corticotropin injection, prevents the effect of corticotropin on protein E synthesis, which is interpreted as evidence that mRNA synthesis is necessary for the stimulation of protein E synthesis. When injected more than 2h after corticotropin, actinomycin D does not prevent corticotropin stimulation of protein E synthesis, but completely blocks corticotropin stimulation of total protein synthesis. This is interpreted as meaning that, after stimulation of mRNA coding for protein E, corticotropin has no effect on the synthesis of protein E. On the other hand, corticotropin stimulation of protein E synthesis persists after hypophysectomy even though it no longer stimulates total protein synthesis. These data suggest that the factor(s) involved in the synthesis of protein E are more stable than those involved in total protein synthesis.

Muscle composition in relation to age and sex

1991 ◽  
Vol 81 (2) ◽  
pp. 249-256 ◽  
Author(s):  
A. M. Forsberg ◽  
E. Nilsson ◽  
J. Werneman ◽  
J. Bergström ◽  
E. Hultman
Keyword(s):  
Protein Content ◽  
Dna Content ◽  
Soluble Protein ◽  
Fat Content ◽  
Age Group ◽  
Soluble Protein Content ◽  
Muscle Composition ◽  
Tissue Weight ◽  
Total Creatine ◽  
Age And Sex

1. A method is described enabling the determination of fat, water, electrolytes, protein, DNA, RNA and total creatine in a single sample of human muscle obtained by the percutaneous needle-biopsy technique. The amino acid content can also be analysed in the same muscle sample. 2. Fifty healthy subjects were studied: 29 between 19 and 40 years of age, 11 between 41 and 60 years of age, and 10 between 61 and 85 years of age. The two groups aged less than 60 years showed only marginal differences in muscle composition, whereas the highest age group showed increases in muscle fat content in relation to tissue weight and decreases in alkali-soluble protein content in relation to both tissue weight and tissue DNA content. Also, potassium, magnesium, total creatine and RNA contents were decreased in this age group when related to tissue DNA content. When alkali-soluble protein was used as a reference base, only magnesium content was decreased. 3. A comparison was also made between female (n = 23) and male (n = 18) subjects in the age groups below 60 years. Differences observed included a higher fat content in female muscle, and an increase in total creatine content in relation to tissue weight. The alkali-soluble protein content was lower per muscle cell in the females when calculated on the basis of DNA content. 4. The results show that in the assessment of muscle constituents, age and sex must be taken into account.

Synthesis of Albumin with Exogenous Mouse-Liver Messenger RNA in a Homologous Cell-Free System

1974 ◽  
Vol 52 (5) ◽  
pp. 429-432 ◽  
Author(s):  
A. J. Faber ◽  
S. H. Miall ◽  
T. Tamaoki
Keyword(s):  
Protein Synthesis ◽  
Total Protein ◽  
Gel Electrophoresis ◽  
Messenger Rna ◽  
Mouse Liver ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
Free System ◽  
Cell Free System ◽  
Membrane Bound

RNA extracted from total and membrane-bound polysomes of mouse liver was capable of directing protein synthesis in a homologous cell-free system in the presence of a 0.5 M KCl ribosomal wash fraction. Analysis of the products by polyacrylamide gel electrophoresis and immunoprecipitation showed that newly formed albumin could account for up to 8% of the total protein synthesized.

Sequential changes in in vivo muscle and liver protein synthesis and plasma and tissue glutamine levels in sepsis in the rat

2001 ◽  
Vol 101 (3) ◽  
pp. 295-304 ◽  
Author(s):  
Michael J. O'LEARY ◽  
Colin N. FERGUSON ◽  
Michael J. RENNIE ◽  
Charles J. HINDS ◽  
John H. COAKLEY ◽  
...  
Keyword(s):  
Protein Synthesis ◽  
Protein Content ◽  
Total Protein ◽  
Muscle Protein ◽  
Muscle Protein Synthesis ◽  
Total Protein Content ◽  
Sham Operation ◽  
Liver Protein ◽  
Male Wistar Rats

We have investigated sequential changes in skeletal muscle and hepatic protein synthesis following sepsis, and their relationship to changes in circulating and tissue glutamine concentrations. Male Wistar rats underwent caecal ligation and puncture (CLP) or sham operation, with starvation, and were killed 24, 72 or 96 h later. A group of non-operated animals were killed at the time of surgery. Protein synthesis was determined using a flooding dose of l-[4-3H] phenylalanine, and glutamine concentrations were measured by an enzymic fluorimetric assay. Protein synthesis in gastrocnemius muscle fell in all groups. Gastrocnemius total protein content was reduced after CLP and at 72 and 96 h after sham operation. After CLP, protein synthesis was lower at 24 h, and total protein content was lower at 72 and 96 h, than in sham-operated animals. CLP was associated with increased liver protein synthesis at all time points, whereas there was no change after sham operation. Liver protein content did not change after CLP, but was lower at 72 and 96 h after sham operation than in non-operated animals. Plasma glutamine concentrations were reduced at 24 h after sham operation, and at 72 and 96 h after CLP. Muscle glutamine concentrations were reduced in all groups, with the decrease being greater following CLP than after sham operation. In the liver, glutamine concentrations were unchanged after CLP, but increased after sham operation. In rats with sepsis, decreases in muscle protein synthesis and content are associated with markedly reduced muscle glutamine concentrations. Plasma glutamine concentrations are initially maintained, but fall later. In liver, protein synthesis is increased, while glutamine concentrations are preserved. These results support a peripheral-to-splanchnic glutamine flux in sepsis.

Labelling of Proteins by Isolated Rat Liver Nuclei

1972 ◽  
Vol 50 (2) ◽  
pp. 190-199 ◽  
Author(s):  
K. M. Anderson ◽  
M. Slavik ◽  
O. P. Elebute
Keyword(s):  
Protein Synthesis ◽  
Rat Liver ◽  
Nuclear Protein ◽  
Sucrose Solution ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Rat Liver Nuclei ◽  
Teleological Argument ◽  
Liver Nuclei ◽  
Triton X

Rat liver nuclei, isolated in hypertonic sucrose solution and washed with Triton X-100, incorporate radioactive amino acids into hot trichloroacetic acid insoluble materials.Optical and biochemical evidence of nuclear purity is presented. The temperature-dependent incorporation continued for 20–30 min, and was proportional to the concentrations of both nuclear protein between 0.5–1.5 mg/ml, and radioactive amino acid. The radioactive product was degraded by pronase, and a number of inhibitors reduced incorporation, but only if present at [Formula: see text]. Proteins extracted from labelled nuclei and microsomes and examined by sodium dodecyl sulfate–polyacrylamide gel electrophoresis at pH 7.2 exhibited different patterns of radioactivity. This provides further support for the concept of protein synthesis intrinsic to rat liver nuclei.A teleological argument for the function of nuclear protein synthesis is discussed.

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