EFFECTS OF THYROXINE AND PROLACTIN ON COLLAGEN BREAKDOWN IN THE THIGH BONE AND TAIL FIN OF THE RANA CATESBEIANA TADPOLE

Injection of ¹⁴C-proline into the tadpole causes labeling of protein in the collagen fraction of the thigh bone and tail fin. The radioactivity of the ¹⁴C-hydroxyproline residue is about 26% of the total radioactivity in the ¹⁴C-labeled protein of the collagen fraction in the thigh bone as well as in the tail fin. In ¹⁴C-proline-loaded tadpoles into which prolactin has been injected, the radioactivity in the collagen fraction in these tissues is markedly higher than that in control animals. In thyroxine-treated tadpoles, the ¹⁴C-radioactivity of the collagen fraction in the thigh bone is always higher than that of the controls, but it is markedly low in the tail fin. During the incubation of thigh bone and tail fin isolated from ¹⁴C-proline-loaded tadpoles, low molecular weight materials containing ¹⁴C-hydroxyproline are released from the ¹⁴C-labeled protein of these tissues. The rate of ¹⁴C-hydroxyproline release, which represents the rate of collagen breakdown, is higher in thigh bone and tail fin isolated from thyroxine-treated tadpoles and is markedly lower in these tissues isolated from prolactin-treated tadpoles than in those isolated from controls. In these tissues, the high rate of collagen breakdown in thyroxine-treated tadpoles is reduced by prolactin injection.     

HORMONES CONTROLLING COLLAGEN SYNTHESIS DURING METAMORPHOSIS IN THE THIGH BONE OF THE TADPOLE OF RANA CATESBEIANA

The rate of ¹⁴C-proline incorporation into collagen in the thigh bone of the Rana catesbeiana tadpole was determined in vitro. Intraperitoneal injection of bovine prolactin caused an increase in the rate of collagen synthesis during the premetamorphic stages (stages 12–16) and the early metamorphic stage (stage 18), but it exerted no effect on collagen synthesis in the metamorphic stages (stages 20–25). On the other hand, injection of growth hormone stimulated the rate of collagen synthesis in the metamorphic stages and caused a slight increase in the premetamorphic stages. When a tadpole in the early premetamorphic stages (stages 12–14) was kept in 5 × 10₋₈ M thyroxine solution for several days, the rate of collagen synthesis became higher than that in the bone of the control animal. The rate of collagen synthesis was not enhanced by prolactin in the thyroxine-treated tadpole, but was stimulated by growth hormone, even when the thyroxine-treated animal remained in the premetamorphic stages. With the treatment of the tadpole by thyroxine, prolactin-sensitivity seems to be reduced, and growth hormone-sensitivity becomes apparent.     

DEVELOPMENTAL EFFECTS ON PROTEIN SYNTHESIS RATES IN REGIONS OF THE CNS IN VIVO AND IN VITRO

Abstract— Protein synthesis rates have been determined quantitatively in several regions of the nervous system of rats of various ages. The developmental changes in these regions are generally similar with a high rate maintained from several days before birth to about 4 days of age (1.9–2.1% h⁻¹). A decline in the rate ensues thereupon which continues till approx 30 days of age, whence the curve flattens though continuing slowly downward with increasing age. In the young three regions, cerebellum, pineal and pituitary, exhibit exceptionally higher rates (40–50%) than the cerebral hemispheres, pons-medulla, mid brain or cord, which all display curves of similar magnitude and shape. While the rate in the cerebellum eventually declines with age to within 10% of the rate in cerebral hemisphere, rates in the pineal and pituitary though decreasing remain far above (100%) rates in cerebral hemisphere even in adults.
The rate in vitro for slices of cerebellum follows a pattern similar to that shown previously for cerebral hemispheres: in the very young rates are 70–80% of the in vivo value but decline much more rapidly with age and in adult represent only 10–15% of the rate in vivo.
A markedly different pattern is seen in whole (unsliced) pituitaries wherein in vitro rates parallel in vivo rates with increasing age at approx 70–80% of the in vivo rate. Pineals appear to follow a similar pattern.     

EFFECT OF PROLACTIN ON THE TADPOLE TAIL FIN. I. STIMULATORY EFFECT OF PROLACTIN ON THE COLLAGEN SYNTHESIS OF THE TADPOLE TAIL FIN

Bovine prolactin stimulated the growth of connective tissues both in the tail fins and in other regions of the tadpole tail. Correlated with the morphological effect of the hormone on the tadpole tails, protein synthesis in tail fins was promoted about 2 times by prolactin. Experiments performed to determine the kind of protein, the synthesis of which was stimulated by prolactin, revealed that the hormone specifically enhanced collagen synthesis about 40 folds as compared to untreated animals.     

EFFECTS OF ADDED SUBSTANCES ON RNA AND PROTEIN SYNTHESIS IN IMMATURE NEURONS

Abstract— A newly described method for the isolation of morphologically intact neurons from newborn rat brain was used to study the influence of inhibitors and neuroactive substances on RNA and protein synthesis in these cells in vitro . Incorporation of [¹⁴C]-uridine into RNA and [³H]leucine into protein proceeded rapidly and continued up to 3 h. When the incorporation mixture was chased at 20 min with an excess of nonradioactive uridine and leucine, hardly any degradation of labelled RNA was noted during the following 2 h 40 min. In contrast, the specific radioactivity of proteins decreased by 22 per cent indicating turnover of cellular proteins.
Incorporation of labelled leucine into protein was markedly inhibited in the presence of NaF and cycloheximide but not affected in the presence of chloramphenicol or pancreatic RNase. A mixture of ATP + GTP depressed the incorporation by 38 per cent. The responses to ATP + GTP and RNase indicated that the incorporation system was typically cellular. Acetylcholine, γ-aminobutyrate, noradrenaline and phenylalanine in the incubation medium depressed the incorporation of labelled uridine into RNA by 10–30 per cent and 5-hydroxytryptamine by 75 per cent. Acetylcholine, γ-aminobutyrate and noradrenaline had no effect on protein synthesis, while 5-hydroxytryptamine and phenylalanine inhibited the incorporation by 60–80 per cent. Testosterone and prednisolone depressed both RNA and protein synthesis while thyroxine caused slight but non-significant stimulation.     

THE EFFECTS OF INHIBITORS OF RNA AND PROTEIN SYNTHESIS ON CHLOROPLAST STRUCTURE AND FUNCTION IN WILD-TYPE CHLAMYDOMONAS REINHARDI

Wild-type cells of the unicellular green alga Chlamydomonas reinhardi have been grown for several generations in the presence of rifampicin, an inhibitor of chloroplast DNA-dependent RNA polymerase, spectinomycin and chloramphenicol, two inhibitors of protein synthesis on chloroplast ribosomes, and cycloheximide, an inhibitor of protein synthesis on cytoplasmic ribosomes. The effects of cycloheximide are complex, and it is concluded that this inhibitor cannot give meaningful information about the cytoplasmic control over the synthesis of chloroplast components in long-term experiments with C. reinhardi. In the presence of acetate and at the appropriate concentrations, the three inhibitors of chloroplast protein synthesis retard growth rates only slightly and do not affect the synthesis of chlorophyll; however, photosynthetic rates are reduced fourfold after several generations of growth. Each inhibitor produces a similar pattern of lesions in the organization of chloroplast membranes. Only rifampicin prevents the production of chloroplast ribosomes.     

饥饿和冬眠期牛蛙胃肠胰系统内分泌细胞的变化

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THE MORPHOLOGICAL EFFECTS OF PROTEIN SYNTHESIS INHIBITION IN MARSILEA

Marsilea vestita and M. drummondii were grown in sterile cultures to which concentrations of the protein synthesis inhibitors, 2-thiouracil (10 mg/liter) and 5-fluorouracil (1 mg/liter) had been added. When young sporelings are grown in a solution of thiouracil at optimum concentration, there is an inhibition of the rate of leaf formation, a retardation of the leaf heteroblastic series, and all leaves develop as land forms. When thiouracil is added to plants which are already producing typical adult, quadrifid leaves, the effects depend on whether the treated plants are water or land forms. Plants which are typically water forms convert to land forms. After treatment successive leaves develop typical sunken stomata on both leaf surfaces. The tissues of the rhizome, root and petiole are more compact and, in general, the cells of the plant have thicker walls. Vascular patterns are not changed, though the size of the rhizome, root and petiole may be reduced. Plants which are typically land forms are less affected than the water forms, but they show a small reduction in apex volume and an apparent reversion of the leaflet number from the typical quadrifid leaf to a trifid, bifid, or single lamina condition. In both land and water forms apical dominance may be broken by treatment with 10 mg/liter thiouracil or 1 mg/liter fluorouracil and numerous lateral branches develop. Higher concentrations (15–25 mg/liter of thiouracil) may result in abnormal development of lateral axillary buds, petiole bases and leaflets. The meristems of the plant are differentially sensitive to thiouracil; leaflet meristems are most sensitive, the root meristems are the least sensitive. It appears that a true reversion to juvenile leaf development need not occur even though protein synthesis and the volume of the apex are reduced. The development of the land or water form in Marsilea appears to depend on rate of growth. Hence inhibition of the growth of typical water forms, through inhibition of protein synthesis, causes a shift in development toward the morphology typical of land forms.     

DNA SYNTHESIS AND CELL TURNOVER IN RANA PIPIENS TADPOLE LIVER DURING SPONTANEOUS METAMORPHOSIS1

The relationship of DNA synthesis and cellular turnover to biochemical differentiation during metamorphosis of R. pipiens liver was investigated. Average DNA/cell was constant at 11.6 pg/ nucleus through stage XXV; but increased during juvenile growth; during metamorphosis stages, changes in total DNA content must correspond to changes in cell number. Rates of DNA synthesis were estimated by rates of ³H-thymidine incorporated into the acid-precipitable fractions, corrected for both precursor uptake into the acid-soluble pool, and for endogenous thymine pool size. DNA content increased steadily from premetamorphosis until late prometamorphosis; at preclimax stages XVIII and XX there were two successive decreases in DNA content of approximately 30%. Fluctuations in synthesis rates preceded corresponding fluctuations in content; DNA synthesis was maximal at stages XVI and XVIII, decreased nearly ten-fold at metamorphic climax, and then gradually rose again during late climax stages. The size of the endogenous thymine pool increased transitorily during spontaneous metamorphosis corresponding to a stage of maximal DNA synthesis. These results indicate that both DNA synthesis and cellular turnover play a significant role in determining net DNA synthesis rates and content during metamorphosis. Metamorphosis of the tadpole liver appears to be associated with both proliferation and cellular death, perhaps a replacement of “larval” by “adult” cells. Metamorphosis of the liver cannot be occuring in a “fixed population of cells” as is commonly assumed. An interpretation of the population dynamics of the metamorphic liver is presented.     

THE EFFECTS OF PHENYLALANINE ON AMINO ACID METABOLISM AND PROTEIN SYNTHESIS IN BRAIN CELLS IN VITRO1

Incubation of brain cell suspensions with 14 mM-phenylalanine resulted in rapid alterations of amino acid metabolism and protein synthesis. Both thc rate of uptake and the final intracellular concentration of several radioactively-labelled amino acids were decreased by high concentrations oi phenylalanine. By prelabelling cells with radioactive amino acids, phenylalanine was also shown to effect a rapid loss of the labelled amino acids from brain cells. Amino acid analysis after the incubation of the cells with phenylalanine indicated that several amino acids were decreased in their intracellular concentrations with effects similar to those measured with radioisotopic experiments (large neutral > small and large basic > small neutral > acidic amino acids). Although amino acid uptake and efflux were altered by the presence of 14 mwphenylalanine, little or no alteration was detected in the resulting specific activity of the intracellular amino acids. High levels of phenylalanine did not significantly altcr cellular catabolism of either alanine, lysine, leucine or isoleucine. As determined by the isolation of labcllcd aminoacyl-tRNA from cells incubated with and without phenylalanine, there was little or no alteration in the level of this precursor for radioactive alanine and lysine. There was, however, a detectable decrease in thc labelling of aminoacyl-tRNA for leucine and isoleucine. Only aftcr correcting for the changes of the specific activity of the precursors and thcir availability to translational events, could the effects of phenylalanine on protein synthesis be established. An inhibition of the incorporation into protein for each amino acid was approximately 20%.     

PVA和PEG(6000)预处理对大豆胚轴生长、蛋白质合成和ATP含量的影响

在低温吸胀阶段,经PVA(聚乙烯醇)和PEG(聚乙二醇6000)预处理的大豆胚轴蛋白质合成和ATP含量均比对照高。在萌发阶段,胚轴生长增快,蛋白质合成明显加快,ATP迅速被消耗,而对照胚轴则相反。试验结果表明,预处理大豆种子萌发和生长与其蛋白质合成、ATP水平和消耗能力有密切关系。     

DNA SYNTHESIS IN RANA PIPIENS TADPOLE LIVER DURING TRIIODOTHYRONINE-INDUCED METAMORPHOSIS1

The relationship of DNA synthesis and cellular turnover to biochemical differentiation during Ts-induced metamorphosis of R. pipiens liver was investigated. Rates of DNA synthesis were estimated by rates of 3 H-thymidine incorporation into the acid-precipitable fractions, corrected for both precursor uptake into the acid-soluble pool, and for endogenous thymine pool size. During T3 -induced metamorphosis, periods of DNA synthesis and fluctuations in DNA content preceded expression of biochemical differentiation as measured by the enzyme arginase, and fluctuations in synthesis rates preceded corresponding fluctuations in content. The earliest response to T3- , was a 50% decrease in liver DNA, followed by increases in thymidine incorporation at 16 hr, 2 days, and 5-8 days. The size of the endogenous thymine pool was not significantly altered by T3 These results indicate that both DNA synthesis and cellular turnover play a significant role in determining net DNA synthetic rates and content during metamorphosis. Expression of thyroxin-induced development of the tadpole liver appears to be associated with both proliferation and cellular death, and metamorphosis of the liver cannot be occurring in a “fixed population of cells.”     

饲料中添加胆汁酸对牛蛙生长性能、体组成和营养物质表观消化率的影响

试验研究了饲料中添加胆汁酸对牛蛙Rana (Lithobates) catesbeiana生长性能、体成分和营养物质表观消化率的影响。基础饲料以鱼粉、豆粕为主要蛋白源, 棕榈油为主要脂肪源, 在基础饲料中分别添加0、100、200和300 mg/kg的胆汁酸(含量99.1%)制作4种等氮等能的试验饲料。试验将144只牛蛙初始体重(75.014.23) g随机分为4组, 每组3个重复, 每个重复12只牛蛙, 进行为期8周的生长试验。结果显示: 各试验组牛蛙成活率和摄食率差异不显著(P0.05)。200 mg/kg胆汁酸添加组牛蛙的增重率、特定生长率、饲料效率、蛋白质效率和氮保留率显著高于对照组(P0.05)。在饲料中添加胆汁酸显著降低了牛蛙脏体指数和全体的脂肪含量(P0.05), 但不影响肌肉粗脂肪以及全体和肌肉水分、粗蛋白、粗灰分含量(P0.05)。各处理组间血清总胆固醇、甘油三酯、高密度脂蛋白胆固醇和低密度脂蛋白胆固醇浓度均无显著变化(P0.05), 血清尿素氮含量随着饲料中胆汁酸添加量的增加而降低, 添加300 mg/kg胆汁酸组牛蛙血清尿素氮含量显著低于对照组(P0.05)。血糖水平则随着胆汁酸添加量的增加而先降低后升高, 其中100 mg/kg胆汁酸添加组牛蛙血糖水平最低(P0.05)。100 mg/kg胆汁酸添加组牛蛙肠道脂肪酶活力显著高于对照组和300 mg/kg胆汁酸添加组(P0.05), 与200 mg/kg胆汁酸添加组之间差异不显著(P0.05), 蛋白酶活力与脂肪酶活力有相似的变化趋势, 而淀粉酶活力未受胆汁酸添加水平影响。各处理组饲料营养物质的表观消化率随饲料中胆汁酸添加量的增加而先升高后降低, 其中饲料添加200 mg/kg胆汁酸显著增加了牛蛙对饲料营养物质的表观消化率(P0.05)。结果表明: 在饲料中添加适量的胆汁酸能提高牛蛙对饲料的消化率, 提高饲料效率, 从而促进牛蛙的生长, 同时可促进蛙体脂肪代谢, 降低机体脂肪沉积, 提高牛蛙可食部分比例。根据本试验结果, 建议牛蛙饲料中胆汁酸添加水平为200 mg/kg。