Chick Embryo Heart Cells Research Articles

Influence of insulin on sensitivity to tetrodotoxin of isolated chick embryo heart cells in culture

A progressive evolution of sensitivity to TTX and to Mn 2+ was observed in isolated chick embryo heart cells in culture. TTX-insensitivity progressively develops and affects the whole cell population after 48 h of cultivation. At the same time, spontaneously beating cells become sensitive to Mn ions which inhibit their activity. Insulin prevents the decrease of TTX-sensitivity of the cell population during the 4 to 48 h time of cultivation. It is assumed that the artificial conditions of the tissue culture method are partly responsible for the changes of the cell membrane properties. An improvement of these conditions by adding insulin to the culture medium can inhibit the loss of normal properties such as TTX-sensitivity. Whether insulin acts by influencing the value of the resting potential or by controlling protein synthesis in the cultured cells remain unknown.

Regulation of amino acid transport in chick embryo heart cells. IV. Site and mechanism of insulin action

The regulation of amino transport by insulin in chick embryo heart cells has been studied. Experiments were designed to identify the system(s) of mediation involved in this regulation and to investigate the mechanisms of the hormone action. Results and conclusions based on the adopted experimental approach include the following: 1. 1. Among the four systems (A, ASC, L, Ly +) found to be operative in cardiac cells for the transport of neutral and basic amino acids (Gazzola, G. C., Franchi-Gazzola, R., Ronchi, P. and Guidotti, G. G. (1973) Biochim. Biophys. Acta 311, 292–301), only the A mediation is responsive to insulin. 2. 2. The hormone enhances the activity of this mediation by increasing the maximal velocity ( V) of transport (without substantial changes in K m ) even under conditions of inhibited protein synthesis. 3. 3. The rate of degradation of protein components of the A mediation (as estimated by measurements of transport activity under conditions of inhibited protein synthesis) is decreased when cells are incubated in the presence of insulin. The hormone does not affect this rate for systems ASC, L and Ly +. 4. 4. Cells previously incubated in a medium containing cycloheximide (phase of inhibited translation) subsequently exhibit a net increase of transport activity (A mediation) when transferred into a medium containing actinomycin D (phase of inhibited transcription). The presence of insulin during pre-incubation in cycloheximide has no effect on the subsequent rate of change in transport activity; when added to the second phase, the hormone definitely enchances transport activity. 5. 5. These observations have been interpreted to indicate that insulin modulates the activity of the A transport system by acting at two different sites; at the cell membrane by protecting specific transport proteins against degradation and at a post-transcriptional level by increasing the rate of synthesis of these transport proteins.

25] Production of glass bead-immobilized catecholamines

Porous glass beads were used originally in 1969 for the insolubitization of enzymes. Since that time numerous enzymes, proteins and cofactors have been immobilized on this solid support. The catecholamines, epinephrine, norepinephrine, and isoproterenol when bound to glass beads have been shown to be biologically active on beating chick embryo heart cells in culture, whole chicken embryo hearts, hearts in open-chested anesthetized dogs, cat papillary muscles, and isolated perfused guinea pig hearts, as well as stimulating cAMP formation in glial tumor cells grown in vitro. The immobilized catecholamines have also been used as an affinity system to partially purify the β-adrenergic-receptor adenylate eyelase complex. This chapter describes the methods of derivatizing glass beads and the procedures for coupling biogenic amines to porous glass beads. Some of the procedures outlined in the chapter for derivatizing the glass are modifications of previous procedures.

Interaction of fatty acid and glucose oxidation by cultured heart cells.

Chick embryo heart cells in tissue culture actively oxidize [1-(14)C]palmitate to (14)CO(2). Fatty acid oxidation by cell monolayers was linear with time and increasing protein concentration. The addition of carnitine to the assay medium resulted in a 30-70% increase in the rate of fatty acid oxidation. The specific activity of palmitic acid oxidation did not change significantly with time in culture and was also the same in rapidly proliferating and density-inhibited cell cultures. Addition of unlabeled glucose to the assay medium resulted in a 50% decrease in (14)CO(2) production from [1-(14)C]palmitate. Conversely, palmitate had a similar sparing effect on [(14)C]glucose oxidation to (14)CO(2). Lactate production accounted for most of the glucose depleted from the medium and was not inhibited by the presence of palmitate in the assay. Thus, the sparing action of the fatty acids on glucose oxidation appears to be at the mitochondrial level. The results indicate that although chick heart cells in culture are primarily anaerobic, they can oxidize fatty acid actively.

Regulation of amino acid transport in chick embryo heart cells III. Formal identification of the A mediation as an adaptative transport system

An amino acid transport system, subject to regulation involving a repression-derepression mechanism by its substrate molecules, is operative in chick embryo heart cells (Gazzola G.C.; Franchi R.; Saibene V.; Ronchi P.; Guidotti G.G. (1972) Biochim. Biophys. Acta 266, 407–421). The formal identification of this system with the A mediation has been achieved by studies yielding the following results. 1. 1. The system which exhibits adaptive control is Na + dependent, whereas its regulation (derepression upon incubation of cardiac cells in amino acid-free medium, repression in the presence of pertinent amino acid) is largely Na + independent. 2. 2. Among the four transport systems (A, ASC, L, Ly +) individually discriminated from interfering systems by relatively specific model substrates, only the A mediation showed derepression when cardiac cells were incubated in amino acid-free medium. 3. 3. Among the four model amino acids tested ( i.e. α-(methylamino)isobutyric acid, b(±)-2-aminobicyclo(2,2,1)heptane-2-carboxylic acid, 4-amino-1-guanylpiperidine-4-carboxylic acid, α,α-diethylglycine) only α-(methylamino)isobutyric acid, developed as specific substrate of System A, inhibited the increase in transport activity of cardiac cells incubated under conditions otherwise derepressive. 4. 4. The addition of transport-specific amino acids during incubation under conditions of inhibited protein synthesis increased the rate of degradation of protein components of System A (as estimated by measurements of transport activity). This had no effect on the rate for System ASC and apparently decreased it for Systems L and Ly +. 5. 5. Experiments, showing that adaptive control of transport activity by the A mediation is operative in such mammalian tissues as newborn mouse and rat heart cells, suggest that this regulation is an intrinsic property of muscle tissue.

Regulation of amino acid transport in chick embryo heart cells II. Adaptive control sites for the “A mediation”

The relative roles of transcription and translation of specific mRNA in the adaptive control of amino acid transport by the A mediation in chick embryo heart cells has been investigated, together with the mode of action of interfering amino acids. Results and conclusions based on the adopted experimental approach include the following: 1. Cells previously incubated in amino acid-free medium containing cycloheximide (phase of inhibited translation) subsequently exhibited a net increase of transport activity when transferred into a plain medium, a medium containing actinomycin D (phase of inhibited transcription), or a medium containing repressive concentrations of alanine (as representative amino acid of the A transport system). 2. The presence of alanine during prior incubation in cycloheximide prevented the increase of activity of the A transport system during subsequent incubation in cycloheximide-free actinomycin D-containing medium. Phenylalanine (as representative amino acid of the L mediation) was ineffective. 3. The synthesis of transport proteins of the A system (as estimated by measurements of transport activity) increased with the duration of cell preincubation in cycloheximide approaching equilibrium after about 40 min. The capacity to synthesize proteins rate-limiting to transport (as a measure of specific mRNA present) decreased during incubation of the cells in cycloheximide when actinomycin D or alanine were added. In both cases capacity decreased as a single exponential with comparable half-lives (approx. 170 min). 4. The degradation of transport proteins of the A mediation followed a single exponential with a half-life of about 200 min when cells were incubated with cycloheximide or puromycin. Actinomycin D did not alter the rate of degradation. Accelerated degradation (half-life of about 80 min) was observed when alanine was added to cells incubated in the presence of cycloheximide. 5. Actinomycin D did not promote the synthesis of transport proteins under conditions of derepressed (absence of added amino acids) or repressed (presence of alanine) transport activity of the A mediation.

Inhibition of the Transport of Several Hexoses in Mammalian Cells by Cytochalasin B

Abstract Cytochalasin B inhibited the transport of several hexoses in HeLa, 3T3, HTC rat hepatoma, and chick embryo heart cells. Fifty per cent inhibition of [3H]glucose, [3H]glucosamine, and 2-deoxy[3H]glucose transport occurred with 8 x 10-8 m cytochalasin B. Inhibition of hexose transport was immediate and was immediately and completely reversible when cytochalasin B was removed from the medium after 1 hour of incubation. Cytochalasin B did not prevent the uptake of thymidine, uridine, α-aminoisobutyric acid, or calcium. Hexokinase activity of a HeLa cell sonicate was also unaffected by cytochalasin B. Inhibition of glucose transport appeared to be noncompetitive.

Regulation of amino acid transport in chick embryo heart cells: I. Adaptive system of mediation for neutral amino acids

The regulation of amino acid transport systems in chick embryo heart cells has been studied. Experiments were designed to investigate activity and kinetics of transport for analogue and natural amino acids as a function of time under various in vitro conditions (active and inhibited protein and RNA synthesis, amino acid dependence, transinhibition). Conclusions based on the proposed experimental approach include the following: 1. 1. The transport activity of a group of neutral amino acids (corresponding to those ascribed to the A mediation by Christensen and co-workers, J. Biol. Chem., 238(1963)3686; Adv. Enzymol., 32(1969)1) increases with time when cells or intact hearts are incubated in the absence of added amino acids. This increase is abolished in the presence of puromycin, cycloheximide and actinomycin D. 2. 2. The increase in activity of the A transport system (investigated by aminoisobutyrate and proline as representative amino acids) is prevented by the addition to the incubation medium of amino acids assigned to the same system of mediation on the basis of competition studies. It is not substantially altered by the addition of amino acids pertaining to the L system of mediation or basic amino acids. 3. 3. Transinhibition does not appear of major importance in the regulation of the activity of the A transport system. 4. 4. Kinetic analyses of aminoisobutyrate uptake in which the transport of the analogue by the A and L systems of mediation could be separated stress further that a time-dependent regulation is effective for the sole A transport system (affecting the maximal velocity without substantial changes in K m ) and provide a comprehensive explanation for previously reported changes with time and intracellular substrate concentration of the kinetic parameters governing the uptake of the analogue 9. 5. 5. Provisional mechanisms for the observed adaptive regulation of the activity of the A transport system for neutral amino acids have been considered.

Biologically active catecholamines covalentyly bound to glass beads.

Catecholamines bound covalently to glass beads have been found to have biological activity in several systems. Experimental evidence has been found that immobilized epinephrine and isoproterenol accelerate the heart rate in dogs, chick embryo, and chick heart cells grown in culture, whereas immobilized propranolol results in a decrease in heart rate. Isoproterenol bound to glass beads has also been shown to markedly increase the level of adenosine 3':5'-cyclic monophosphoric acid in glial cells. The effects of the immobilized catecholamines are of longer duration than when the compounds are administered in solution. The present data indicate that the compounds are exerting their action when bound to the beads.

Changes in glucose oxidation during growth of embryonic heart cells in culture.

A rapid and convenient method has been utilized to investigate glucose oxidation during growth of chick embryo heart cells in tissue culture. Primary isolates of chick embryo heart cells showed exponential growth when plated at low densities and exhibited density-inhibited growth as cultures became confluent. The density-dependent growth inhibition of chick embryo heart cells is associated with a marked decrease in the specific activity of glucose oxidation to CO(2). This decrease in glucose oxidation was observed as density increased as either a function of time in culture or as related to initial plating density. The decrease in (14)CO(2) production associated with density-dependent inhibition of growth is due to a marked decrease in activity of the pentose phosphate pathway.

The Effect of Acetylcholine on Single Embryonic Chick Ventricle Cells

SummaryThe effect of acetylcholine on the beating rate of single chick embryo heart cells cultured from 8 to 10 day ventricles was determined using a new television monitor optical system. Acetylcholine had a clear negative chronotropic effect on these cells, with a threshold at 10-8-10-7 g/ml. The slowing in beating rate after the addition of acetylcholine was reversible, and was not associated with any detectable change in the inotropic state of the cells. These findings are consistent with the effect of acetylcholine on non-cultural chick embryo ventricular tissue of the same age.

Insulin effect on amino acid transport in bone: dependence on protein synthesis and Na+.

ARTICLESInsulin effect on amino acid transport in bone: dependence on protein synthesis and Na+TJ Hahn, SJ Downing, and JM PhangTJ Hahn, SJ Downing, and JM PhangPublished Online:01 Jun 1971https://doi.org/10.1152/ajplegacy.1971.220.6.1717MoreSectionsPDF (2 MB)Download PDF ToolsExport citationAdd to favoritesGet permissionsTrack citations ShareShare onFacebookTwitterLinkedInWeChat Previous Back to Top Next Download PDF FiguresReferencesRelatedInformation Cited BySLC38A2 provides proline to fulfil unique synthetic demands arising during osteoblast differentiation and bone formation9 March 2022 | eLife, Vol. 11The role of osteoblasts in energy homeostasis28 August 2019 | Nature Reviews Endocrinology, Vol. 15, No. 11Wnt signaling and cellular metabolism in osteoblasts26 November 2016 | Cellular and Molecular Life Sciences, Vol. 74, No. 9Bone Cell Bioenergetics and Skeletal Energy HomeostasisRyan C. Riddle and Thomas L. Clemens15 February 2017 | Physiological Reviews, Vol. 97, No. 2Magnesium HomeostasisMagnesium HomeostasisDisproportionate alterations in body composition of large for gestational age neonatesThe Journal of Pediatrics, Vol. 138, No. 6A chemically modified tetracycline inhibits streptozotocin-induced diabetic depression of skin collagen synthesis and steady-state type I procollagen mRNABiochimica et Biophysica Acta (BBA) - Molecular Cell Research, Vol. 1402, No. 3The regulation of L-proline transport by insulin-like growth factor-I in human osteoblast-like SaOS-2 cellsPflügers Archiv - European Journal of Physiology, Vol. 432, No. 3Bone mineral density at the metaphysis is specifically reduced in STZ-treated diabetic ratsLife Sciences, Vol. 59, No. 2Osteoporosis: Pathophysiology, prevention, diagnosis, and treatmentDisease-a-Month, Vol. 39, No. 11Insulin and GrowthTetracycline administration increases protein (presumably procollagen) synthesis and secretion in periodontal ligament fibroblasts of streptozotocininduced diabetic ratsJournal of Periodontal Research, Vol. 27, No. 6The effects of dietary protein insufficiency and excess on skeletal healthBone, Vol. 13, No. 4Parathyroid hormone regulation of proline uptake by cultured neonatal mouse osteoblastlike cells3 December 2009 | Journal of Bone and Mineral Research, Vol. 4, No. 1Endocrinology of Bone FormationGlucose transport system in UMR-106-01 osteoblastic osteosarcoma cells: Regulation by insulinCalcified Tissue International, Vol. 45, No. 1Glucose transport in osteoblast-enriched bone explants: Characterization and insulin regulation3 December 2009 | Journal of Bone and Mineral Research, Vol. 3, No. 3Effect of parathyroid hormone on amino acid transport by cultured neonatal mouse calvarial bone cells3 December 2009 | Journal of Bone and Mineral Research, Vol. 3, No. 2Bone mineral density in diabetes mellitusBone, Vol. 9, No. 1The influence of a high-protein, low-carbohydrate diet on bone development in the fetuses of rat dams with streptozotocin-induced diabetes9 March 2007 | British Journal of Nutrition, Vol. 59, No. 1Osteoblastlike cells in a serum-free methylcellulose medium form colonies: Effects of insulin and insulinlike growth factor ICalcified Tissue International, Vol. 40, No. 1The influence of progressive hyperinsulinemia on duodenal calcium absorption in the ratMetabolism, Vol. 36, No. 1Effects of denervation and protein synthesis inhibitors on amino acid transport in the rat diaphragmActa Physiologica Scandinavica, Vol. 123, No. 4Regulation of Bone FormationNew England Journal of Medicine, Vol. 309, No. 2Pathophysiology of the diabetic boneHormonal Modifiers of Mineral Metabolism Other Than Parathyroid Hormone, Vitamin D, and CalcitoninDevelopment of bone mineral loss in insulin-treated diabetes: a 1 1/2 years follow-up study in sixty patientsEuropean Journal of Clinical Investigation, Vol. 11, No. 1Ouabain Effects on Hormonally-Stimulated Bone Resorption and Cyclic AMP Content in Cultured Fetal Rat Bones7 July 2009 | Endocrine Research Communications, Vol. 7, No. 3The regulation of amino acid transport in animal cellsBiochimica et Biophysica Acta (BBA) - Reviews on Biomembranes, Vol. 515, No. 4Characteristics of proline transport into R3230AC mammary tumor cellsBiochimica et Biophysica Acta (BBA) - Biomembranes, Vol. 508, No. 2Effects of neonatal estrogen on in vivo transport of α-aminoisobutyric acid into rat brainExperimental Neurology, Vol. 57, No. 3The effects of neonatal androgenization on the in vivo transport of alpha-aminoisobutyric acid into specific regions of the rat brainBrain Research, Vol. 132, No. 2Osteoporosis: Pathogenesis and TherapyThe blood: Bone disequilibriumCalcified Tissue Research, Vol. 20, No. 1Amino acid transport in isolated rat thymocytes. Effects of divalent cations and ethanol.Journal of Biological Chemistry, Vol. 251, No. 19Effect of fluoride on protein and collagen biosynthesis in rabbit dental pulp in vitroEuropean Journal of Oral Sciences, Vol. 84, No. 5Effects of Diabetes Mellitus on Bone Mass in Juvenile and Adult-Onset DiabetesNew England Journal of Medicine, Vol. 294, No. 5Hormonal Regulation of Cartilage Growth and MetabolismDevelopmental patterns of intestinal transport mechanisms in the chickComparative Biochemistry and Physiology Part A: Physiology, Vol. 54, No. 1Membrane Transport: Its Relation to Cellular Metabolic RatesScience, Vol. 188, No. 4194Regulation of amino acid transport in chick embryo heart cells. IV. Site and mechanism of insulin actionBiochimica et Biophysica Acta (BBA) - Biomembranes, Vol. 356, No. 2Insulin Action in Isolated Rat ThymocytesJournal of Biological Chemistry, Vol. 247, No. 21 More from this issue > Volume 220Issue 6June 1971Pages 1717-1723 Copyright & PermissionsCopyright © 1971 by American Physiological Societyhttps://doi.org/10.1152/ajplegacy.1971.220.6.1717PubMed4253389History Published online 1 June 1971 Published in print 1 June 1971 Metrics

Kinetic analysis of insulin action on amino acid uptake by isolated chick embryo heart cells.

1. Isolated chick embryo heart cells were used to investigate the mode of action of insulin on the transport of three naturally occurring amino acids: l-proline, l-serine and glycine. Initial velocities of uptake were measured over a period of 5min with an 80-fold range of amino acid concentration. Corrections for amino acid diffusion, incorporation into protein and conversion into carbon dioxide were introduced. 2. The uptake processes approximated Michaelis-Menten kinetics within definite ranges of amino acid concentrations. A single transport system for proline and at least two transport systems for serine and glycine were detected. 3. The kinetic effects of insulin on transport systems for the amino acids tested were consistent with an acceleration of the maximal velocity of the process, without substantial changes in substrate concentration for half-maximal transport velocity. 4. These hormonal effects were not essentially altered by the corrections for amino acid incorporation into protein and conversion into carbon dioxide.

Effect of peptides on animal cells in culture: I. Maintenance of beating heart cells

Chick embryo heart cells grown as organ cultures or monolayer cultures maintained beating for varying periods of time in media supplemented with Bacto-Peptone or Proteose Peptone but in the absence of serum. Electron micrographs of these cultures suggested a “normal ultrastructure”, and initial fractionation studies indicated that the factor(s) in either Bacto-Peptone or Proteose Peptone which contributed to the maintenance of beating could be partially purified by gel filtration. Serum, routinely used in tissue culture studies, is not only complex but the response of cells to different lots of serum has been reported to vary significantly [4, 8, 21, 23, 24]. These problems might be resolved by identifying the active component(s) of serum or by substituting less complex materials for the serum component. The latter has been reported by Amborski & Moskowitz [1], and by Amborski et al. [2]. These results suggested that peptides might be used in the place of serum in studies on the growth and development of primary cultures. This report describes the effects of similar peptides on primary cell cultures derived from embryonic chick hearts cultivated under different conditions; in organ culture, in aggregate culture and in monolayer culture.

Amino acid uptake in isolated chick embryo heart cells.

1. The preparation of cell suspensions by treatment of chick embryo hearts with collagenase at various stages of development is described. 2. Measurements of oxygen consumption, incorporation of labelled leucine into protein and accumulation of labelled alpha-aminoisobutyric acid against a concentration gradient indicated a long-lasting viability of the isolated heart cells in vitro; a satisfactory preservation of subcellular structures, including plasma membrane, was assessed by electron-microscopic examination. 3. The rate of alpha-aminoisobutyric acid accumulation by cardiac cells isolated from hearts at different stages of embryological development decreased with aging; insulin stimulated the intracellular accumulation of this amino acid analogue. 4. Insulin increased the uptake by isolated heart cells of several (14)C-labelled naturally occurring amino acids; however, the fraction of amino acid taken up by the cells that was recovered free intracellularly, and therefore the concentration ratio (between intracellular water and medium), was enhanced by the hormone only with glycine, proline, serine, threonine, histidine and methionine. When isolated heart cells were incubated in the presence of a mixture of labelled amino acids, the addition of insulin increased the disappearance of radioactivity from the medium. 5. The general pattern of amino acid transport (in the absence and in the presence of insulin) in isolated cardiac cells was similar to that found in intact hearts, suggesting that the biological preparation described in this paper might be useful for studies of cell permeability and insulin action.

Growth and respiration of monolayer cell cultures of chick embryo heart and skeletal muscle: action of malathion and malaoxon.

SummaryThe respiration of chick embryo heart and pectoral muscle cell cultures was determined using a polarographic respiration chamber designed specifically to measure the oxygen consumption of monolayers of cells. Heart cultures had a mean respiration rate of 215 ± 84 mμmoles O2/hr/106 cells; pectoral muscle cultures had a mean respiration rate of 163 ± 46 mμmoles Oa/hr/106 cells. Malathion 3 × 10−4M reduced the respiration of pectoral muscle cultures to 26% of normal and the respiration of heart cultures to 50% of normal. Malaoxon 3 × 10−4M had no effect upon the respiration of either cell type. Growth studies indicated that malathion was more toxic than malaoxon to the growth of both cell types.

A balloon isolator for culture of heart cells without antibiotics

A sterile environment based on an inflated flexible-film isolator has been successfully employed in the cultivation of isolated chick embryo heart cells. Tissue culture procedures were not limited by the closed system. Whereas bacterial contamination had previously become a serious problem resistant to standard control measures, bacterial contamination was so well controlled by the isolator technique that antibiotics were no longer added to the growth medium. A new method for surface sterilization of embryonated eggs was developed.

Action of isoproterenol on heart cells in tissue culture.

SummaryTrypsin dispersed chick embryo heart cells have been grown in tissue culture to produce rhythmic contraction cell clusters. Isoproterenol, a specific beta adrenergic stimulating drug, has been shown to increase the rate of contraction of these cells. This effect persists for 20-30 minutes and can be removed by washing the cells with fresh control media. Propranolol, a specific beta adrenergic blocking agent, blocks the stimulating action of isoproterenol. It appears likely that stimulation of beta adrenergic receptors of the Ahlquist classification can be used to explain these findings. These receptors apparently develop in the absence of adrenergic nerve endings.Addendum. At a recent symposium on the structure and function of heart muscle. Wollenberger described his unpublished observations (16) on the effects of adrenergic stimulating and blocking drugs on heart cells in tissue culture which are in general agreement with this report.

Glucose uptake in the developing chick embryo heart.

ARTICLESGlucose uptake in the developing chick embryo heartGG Guidotti, L Loreti, G Gaja, and PP FoaGG Guidotti, L Loreti, G Gaja, and PP FoaPublished Online:01 Oct 1966https://doi.org/10.1152/ajplegacy.1966.211.4.981MoreSectionsPDF (2 MB)Download PDF ToolsExport citationAdd to favoritesGet permissionsTrack citations ShareShare onFacebookTwitterLinkedInEmailWeChat Previous Back to Top Next Download PDF FiguresReferencesRelatedInformationCited ByDevelopmental Changes in Membrane Electrical Properties of the HeartInsulin in birds: metabolic effects and possible implications in genetically fat and lean chickensDevelopmental Changes in Membrane Electrical Properties of the HeartSurgar transport in animal cells: The passive hexose transfer systemProgress in Biophysics and Molecular Biology, Vol. 43, No. 1Physiology and pharmacology of developing heart cellsPharmacology & Therapeutics, Vol. 22, No. 1Pacemaker Mechanisms in Myocardial Cells during Development of Embryonic Chick HeartsCardiac myocyte cell surface: Identification and expression of differentiation antigens during chick heart developmentDevelopmental Biology, Vol. 81, No. 2Changes in Membrane Electrical Properties during Development of the HeartD-xylose transport in isolated skeletal muscle of chickens: Effects of insulin and tolbutamideComparative Biochemistry and Physiology Part C: Comparative Pharmacology, Vol. 67, No. 1Effects of insulin and diabetes on 2-deoxy-D-glucose uptake by the isolated perfused rat lung.Journal of Biological Chemistry, Vol. 254, No. 12Developmental control of 2-aminoisobutyric acid transport by 7- and 14-day chick heart cell aggregatesDevelopmental Biology, Vol. 67, No. 1Glucose uptake by chicken embryo hearts at various stages of developmentDevelopmental Biology, Vol. 55, No. 1Carbohydrate MetabolismTHE ONTOGENY OF MAMMALIAN INSULAR FUNCTIONUptake of sorbitol by chick embryo heart cells at various stages of developmentThe Journal of Membrane Biology, Vol. 24, No. 1Membrane transport during development in animalsBiochimica et Biophysica Acta (BBA) - Reviews on Biomembranes, Vol. 415, No. 3Metabolic Effects on Muscular TissueThe Effect of Insulin on Glucose Transport in Muscle CellsRegulation of amino acid transport in chick embryo heart cells. IV. Site and mechanism of insulin actionBiochimica et Biophysica Acta (BBA) - Biomembranes, Vol. 356, No. 2Physical Characteristics and Chemi-osmotic Transformations of Mutarotases from Various SpeciesJournal of Biological Chemistry, Vol. 248, No. 12THE INTERMEDIARY METABOLISM OF BIRDSEvaluation of kinetic parameters for uptake of amino acids by cells: Effect of insulin on the accumulation of aminoisobutyrate and cycloleucine by isolated rat diaphragm muscle and chick embryo heartsBiochimica et Biophysica Acta (BBA) - Biomembranes, Vol. 241, No. 1Endocrine Control of Avian Carbohydrate Metabolism,Poultry Science, Vol. 50, No. 1Studies on MutarotasesJournal of Biological Chemistry, Vol. 245, No. 3Hormonal Control of Fetal Development and MetabolismHormonal Responses in Amphibian MetamorphosisHormones and Transport across Cell MembranesInsulin and Protein Synthesis More from this issue > Volume 211Issue 4October 1966Pages 981-987 Copyright & PermissionsCopyright © 1966 by American Physiological Societyhttps://doi.org/10.1152/ajplegacy.1966.211.4.981PubMed5926589History Published online 1 October 1966 Published in print 1 October 1966 Metrics

Succinic dehydrogenase and cytochrome oxidase activities in cell cultures.

Succinic dehydrogenase and cytochrome oxidase have been assayed in permanent cell lines (HEP 1, HEP 2, and HLM), in short-term cultures of chick embryo heart cells, and in various tissues. Their activities in different cells are compared by relating them to deoxyribonucleic acid. They are very low in HEP 1, HEP 2, and HLM cells by comparison with the activities in any normal tissues examined. All the succinic dehydrogenase was shown to be located in the mitochondria of the permanent cell lines by staining with tetrazolium derivatives. Both enzymes were more active in tissues of 19-day chick embryos than in those of 11- or 14-day embryos. The increasing activities found during normal development were quickly curtailed or reversed when heart cells were grown as monolayer cultures. Digitonin-treated mitochondria produced preparations with much higher activities of cytochrome oxidase than untreated samples. Activities measured in this way were again very much lower in HEP 1, HEP 2, and HLM cells than in the normal tissues. From the derived ratio of cytochrome oxidase:succinic dehydrogenase, it was apparent that cytochrome oxidase is diminished to a greater extent than succinic dehydrogenase in both permanent cell lines and short-term cultures, by comparison with the corresponding activities in embryonic and adult tissues. The features common to the metabolism of proliferating cells in vitro and malignant cells are discussed.