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Biomedical subjects

K Wildenthal

Publications and source records attributed to K Wildenthal.

At least 19 recordsLinked to original sources
PubMed1986

Disproportionate reduction of actin synthesis in hearts of starved rats.

We examined the synthesis of proteins in rat myocardium after starvation. Rates of total protein synthesis in myofibrillar and nonmyofibrillar fractions of myocardium of starved animals were reduced similarly (to 70-80% of the rates in hearts of fed animals, p less than 0.002), but rates of synthesis of some individual proteins were affected discoordinately. Radiolabeled proteins from atrial and ventricular explants, separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, revealed that starvation for 2 days reduced the rate of cardiac actin synthesis to 26-38% of control levels, while the rate of myosin heavy chain synthesis in the same hearts was only moderately reduced (74-80% of control levels). This starvation-induced reduction in actin synthesis could be accounted for at least in part by disproportionately decreased levels of actin mRNA in starved hearts, as revealed by Northern blot hybridization and by in vitro translation analysis. The dramatic decrease in cardiac actin synthesis was rapidly reversible, and actin synthesis returned to normal after a single day of refeeding. The selective reduction of actin synthesis after starvation was specific for the heart: rates of myosin heavy chain and actin synthesis in skeletal muscles (soleus and extensor digitorum longus) were coordinately reduced in response to starvation. To our knowledge, this is the first example of such dramatic discoordinate regulation of myofibrillar protein synthesis in response to a physiological stimulus.

Actins↗
PubMed1986

Lysosomal responses of fetal mouse hearts recovering from anoxia and substrate depletion.

Recovery from a 1 h period of anoxia and substrate deprivation is accompanied by a marked lysosomal response in myocytes of fetal mouse hearts maintained in organ culture. Two classes of subcellular vacuoles form within 5 to 15 min of recovery. One appears to provide lysosomal enzymes for degradation of subcellular particles, while the other segregates organelles within the cytoplasm of the injured myocyte. When the two populations fuse with each other, the degradation of sequestered organelles appears to commence. After 6 h of recovery, intravacuolar degradation appears complete, and the injured myocytes are morphologically indistinguishable from control cells, demonstrating that the breakdown of the partitioned cell organelles is quite efficient. The process can proceed, albeit at a reduced rate, while protein synthesis is inhibited, since cycloheximide only modestly interferes with recovery after reoxygenation. The present results demonstrate that the fetal mouse heart subjected to conditions that simulate some important aspects of ischemia is an excellent model to examine the role of lysosomes during recovery from sublethal injury.

Animals↗
PubMed1986

Influence of agents that alter lysosomal function on fetal mouse hearts recovering from anoxia and substrate depletion.

Recovery of fetal mouse heart myocytes from oxygen and substrate deprivation for 1 h is accompanied by complicated lysosomal and non-lysosomal vacuolar responses which can be subdivided temporally into four distinct phases that include production of lysosomal dense bodies; segregation of damaged subcellular organelles into vacuoles that initially lack lysosomal enzymes; delivery of lysosomal enzymes to these vacuoles through fusion with dense bodies, transforming them into lysosomal autophagic vacuoles and degradation of the sequestered organelles. These events are normally completed within 6 h of the resupply of oxygen and substrate. The progression of these events is influenced significantly by pharmacological interventions that alter lysosomal properties. Chloroquine inhibits all aspects of the lysosomally-related processes as well as the sequestration phase during recovery. Leupeptin delays the lysosomal degradation, presumably by slowing proteolysis. Hydrocortisone permits the engulfment phase and the appearance of lysosomal dense bodies but appears to prevent or postpone the delivery of lysosomal enzymes to many of the large vacuoles and to delay the degradation of sequestered organelles. These observations reveal that segregation of damaged organelles and lysosomally-mediated degradation of these subcellular structures are important events during recovery from ischemic-like injury, and that agents that interfere with normal lysosomal function can prevent or delay some or all of the lysosomal responses that are involved in the recovery process.

Animals↗
PubMed1986

Effects of glucocorticoid treatment on cardiac protein synthesis and degradation.

We treated rats with dexamethasone (DEX, 1 mg . kg-1 . day-1) and examined the effects of this glucocorticoid on heart protein metabolism using atrial explant and Langendorff perfusion preparations. Fasted rats treated with DEX for 2 days had significantly lower body weights (92% of control, P less than 0.001) and larger hearts (106% of control, P less than 0.005) than fasted control animals. Protein and RNA concentrations remained constant. In atrial explants, DEX treatment produced a 19% increase in protein synthesis (P less than 0.001) and a 13% increase in protein degradation (P less than 0.002). In Langendorff-perfused hearts, DEX treatment caused a 36% increase in protein synthesis (P less than 0.02), while protein degradation was 8% above control (P greater than 0.05). Thus, in contrast to their catabolic effects on skeletal muscle, glucocorticoids are anabolic on the heart. The increased accumulation of total cardiac protein during early glucocorticoid administration is mediated entirely via increased rates of synthesis.

Animals↗
PubMed1985

The role of lysosomes and microtubules in cardiac protein degradation.

The mechanisms and regulatory factors involved in cardiac proteolysis are incompletely understood. Agents that interfere with lysosomal function (e.g., chloroquine, leupeptin, methyladenine) cause a 25-30% reduction in the overall rate of protein degradation. In the same hearts, however, the rate of myosin breakdown remains unchanged. Disaggregation of micro-tubules with colchicine is accompanied by a 15% reduction in the rate of degradation of total protein and of myosin. In the same hearts, the degradation of "organellar" protein, including mitochondrial cytochromes, is reduced by over 30%. Thus, it appears that the degradation of different classes of cardiac proteins may be accomplished and regulated by different processes. Lysosomes are important in overall proteolysis, but appear not to be involved in the regulation of myosin breakdown. Microtubules are also involved in the proteolytic process, and appear to be especially important for the breakdown of proteins from mitochondria and perhaps other organelles.

Animals↗
PubMed1985

The role of lysosomes in the degradation of myofibrillar and non-myofibrillar proteins in heart.

Lysosomes are presumed to be involved in protein degradation in heart, but their exact role is poorly understood. Several interventions that are known to alter cardiac proteolysis (e.g., insulin) also produce lysosomal changes that might account for the observed changes in protein degradation; but many other interventions appear not to do so. Agents that interfere with lysosomal function (e.g., sucrose, chloroquine, methyladenine, leupeptin) cause a 25% reduction in the rate of degradation of total protein in fetal mouse hearts in organ culture; however, in the same hearts the rate of degradation of myosin and other myofibrillar proteins remains unchanged. Thus, it appears that lysosomes are involved in cardiac proteolysis, but may not play a rate-limiting or regulatory role in many circumstances. The regulation of proteolysis by insulin appears to involve non-lysosomal pathways in addition to any lysosomal alterations it may cause. Furthermore, the initial cleavage of myofibrillar proteins appears no to be dependent on normal lysosomal function.

Adenine↗
PubMed1983

Inhibition of cardiac proteolysis by colchicine. Selective effects on degradation of protein subclasses.

1. The effect of colchicine (2.5 microM) on cardiac protein turnover was tested with foetal mouse hearts in organ culture. 2. Colchicine had no effect on protein synthesis, but inhibited total protein degradation by 12-18%. Lumicolchicine, which lacks colchicine's ability to disaggregate microtubules, but shares its non-specific effects, did not alter protein degradation. 3. The colchicine-induced inhibition of protein degradation was accompanied by significant changes in cardiac lysosomal enzyme activities and distribution. 4. Colchicine inhibited the degradation of organellar proteins, including mitochondrial cytochromes, more than that of cytosolic proteins. 5. Colchicine decreased the rate of myosin degradation and the rate of proteolysis of the total protein pool to a similar extent. Since the regulation of myosin degradation does not involve lysosomes, this suggests that colchicine affects non-lysosomal as well as lysosomal pathways. 6. Release of branched-chain amino acids from colchicine-treated hearts was disproportionately decreased, suggesting that colchicine increased their metabolism. 7. It is concluded that colchicine, via its actions on microtubules, exerts important inhibitory effects on cardiac proteolysis. Colchicine is especially inhibitory to the degradation of organellar proteins, including mitochondrial cytochromes. Its inhibitory effects may be mediated in part via lysosomal mechanisms, but non-lysosomal mechanisms are probably involved as well.

Amino Acids↗
PubMed1983

Mechanisms of degradation of myofibrillar and nonmyofibrillar protein in heart.

The degradation of cardiac proteins is known to be altered by many physiological and pathological interventions, but the precise intracellular processes that regulate proteolysis and the relative roles of different proteolytic pathways in degrading different classes of protein remain poorly understood. Agents that interfere with lysosomal function produce major decreases in total protein breakdown; thus, lysosomes and lysosomal proteinases seem to be important in proteolysis. However, these same agents cause no change in the degradation of myofibrillar proteins, suggesting that this class of proteins is not dependent on lysosomal pathways for its turnover.

Animals↗
PubMed1983

Changes in cardiac cathepsin B activity in response to interventions that alter heart size or protein metabolism: comparison with cathepsin D.

The specific activity of cardiac cathepsin B is significantly decreased by starvation and corticosteroid treatment in vivo, and by exposure of the heart in vitro to insulin, hydrocortisone and cycloheximide. Increases in cathepsin B activity occur following isoproterenol-induced cardiac damage in vivo and exposure in vitro to sucrose. Cathepsin B activity in heart is not changed during normal aging or in thyrotoxicosis. These responses are different from simultaneous changes in cardiac cathepsin D activity in several instances (starvation, corticosteroid treatment, aging and thyrotoxicosis). In the past, measurements of cathepsin D activity in heart have sometimes been considered to be representative of lysosomal proteinase activity in general and used as an index of cardiac lysosomal proteolytic capacity. The present results suggest that changes in cathepsin D do not necessarily reflect alterations in other lysosomal proteinases and may not serve as a valid indicator of overall lysosomal proteolytic capacity under all conditions.

Age Factors↗
PubMed1983

Influence of chlorpromazine on lysosomal alterations during myocardial ischaemia.

Ligation of the circumflex artery of anaesthetised, open-chest rabbits caused a progressive increase in nonsedimentable cathepsin D activity in severely ischaemic myocardium and an anatomical redistribution of the enzyme from lysosomes into the cytosol, along with progressive ultrastructural signs of cellular damage and necrosis. Chlorpromazine pretreatment (15 mg X kg-1 intravenously) reduced the increase in nonsedimentable cathepsin D activity slightly, but no appreciable protective effect on the anatomical redistribution of the enzyme or the development of ultrastructural signs of necrosis could be detected. It is concluded that in this experimental model of myocardial infarction, high concentrations of chlorpromazine have a mild stabilising action on lysosomes, but the drug has minimal if any effect in protecting the heart from ischaemic damage.

Animals↗
PubMed1983

Direct anabolic effects of thyroid hormone on isolated mouse heart.

The direct effects of L-and D-triiodothyronine (T3) on cardiac protein metabolism were investigated using fetal mouse hearts in organ culture. This model allowed the production of "thyrotoxicosis" in isolated hearts in vitro in the absence of the usual systemic metabolic and hemodynamic effects of thyroid hormones. Hearts were studied during the first 24 h of T3 exposure in culture, before changes in beating rate due to T3 occurred. Phenylalanine release was decreased by 26 +/- 2.3% (P less than 0.001) by the optimal concentrations of T3 (10(-7) to 10(-6) M). Changes were similar in the presence or absence of insulin. D-T3 was also anabolic, decreasing phenylalanine release by 24 +/- 2.5% (P less than 0.001) at concentrations of 10(-6) to 10(-5) M. The L-isomer increased protein synthesis by 23 +/- 6.8% (P less than 0.05) and decreased protein degradation, as measured by phenylalanine release in the presence of cycloheximide, by 5 +/- 1.6% (P less than 0.01). The D-isomer also increased protein synthesis but had no measurable effect on protein degradation. We conclude that thyroid hormones can exert direct anabolic effects on heart in the absence of systemic hemodynamic and metabolic changes. These effects are mediated primarily through an acceleration of the rate of protein synthesis; in the case of L-T3, a small inhibition of proteolysis may also occur.

Amino Acids↗
PubMed1981

Intracellular disruption of rat heart lysosomes by leucine methyl ester: effects on protein degradation.

Perfusion of rat hearts with Krebs--Henseleit medium containing 10 mM L-leucine methyl ester leads to swelling of lysosomes and loss of lysosomal integrity within 30-60 min. No morphological changes can be detected in the nuclei, mitochondria, sarcoplasmic reticulum, or Golgi complex as a result of the treatment with leucine methyl ester, and the hearts continue to beat normally during the treatment period. Homogenates of rat hearts perfused with the methyl ester exhibit a decrease in the sedimentability of cathepsin D activity compared to controls, thus providing additional evidence for a loss of lysosomal integrity. Swelling and disruption of the lysosomes presumably occurs because of the extensive accumulation of leucine within the organelles resulting from the intralysosomal hydrolysis of the freely permeating methyl ester. The lysosomal dysfunction that occurs with exposure to leucine methyl ester produces a 30% decrease in cardiac protein degradation. These results provide an estimate of the contribution of lysosomes to total protein degradation in the rat heart, and they also suggest that the enzymes released as a result of lysosomal disruption are relatively inactive in hydrolyzing cellular constituents under the perfusion conditions used here. The use of amino acid methyl esters to produce rapid, specific loss of lysosomal integrity in situ provides an approach to the study of lysosomal function in intact cells.

Animals↗
PubMed1981

Lysosomal alterations in heart, skeletal muscle, and liver of hyperthyroid rabbits.

Daily administration of L- or D-thyroxine for 1 week produced hypertrophy of the heart and atrophy of skeletal muscle and liver. The myocardial hypertrophy was accompanied by a rise in the activity of cathepsin D but not of cathepsin B; this was correlated with an increase in cathepsin-D-rich interstitial cells while the number of cathepsin-D-positive lysosomes in myocytes was decreased, as assessed from immunohistochemistry. In atrophying skeletal muscle (soleus and tibialis anterialis), large increases in the activities of cathepsins B and D were present. Immunohistochemical localization of cathepsin D revealed that in thyrotoxic striated muscle cells this acid proteinase had become localized diffusely in the paranuclear myoplasm. The atrophying liver of thyrotoxic rabbits also developed large increases in cathepsin D activity, but in this organ the increase was correlated with an increased number of cathepsin-D-positive secondary lysosomes without diffuse extralysosomal deposits. These observations indicate that changes in lysosomes and lysosomal enzyme activities elicited by thyrotoxicosis are tissue-specific. In some organs, the changes may be associated with net changes in protein balance or with tissue injury, but the exact functional significance of the lysosomal alterations remains uncertain.

Animals↗