Culture of the terminally differentiated ventricular cardiac muscle cell

Claycomb, William C.; Burns, Alvin C.; Shepherd, R. E.

doi:10.1016/0014-5793(84)80330-0

Cited by 19 publications

(4 citation statements)

References 19 publications

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“…Transcription mepreparation of adult rat cardiac myocytes [26,27] with diated by CREB regulates diverse cellular responses minor modifications. Four 200-250 g female Spragueincluding intermediary metabolism and cell proliferation.…”

Section: Introductionmentioning

confidence: 99%

Comitogenic effect of catecholamines on rat cardiac fibroblasts in culture

Leicht

Greipel

Zimmer

2000

Cardiovascular Research

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Objective: We studied the ability of norepinephrine and of other catecholamines to affect the proliferation of cardiac fibroblasts isolated from adult rat hearts. Furthermore, we investigated the possible adrenergic receptor involved in this process. Methods: Norepinephrine (NE), phenylephrine (PE), isoproterenol (ISO), forskolin (FO), epidermal growth factor (EGF), platelet-derived growth factor AA (PDGF-AA) and specific inhibitors of the a -, a -, b -and b -adrenoceptors and of the protein kinase A (PKA) were applied 1 2 1 2 to cardiac fibroblasts in culture. Cell number was measured by use of a Coulter Counter. Activation of the cAMP response element binding protein (CREB) was measured by Western blotting and subsequent use of a phospho-specific antibody. Activation of the p42-and MAPK the p44-mitogen activated protein kinase (p42 / p44 ) was assessed by detection of phosphorylation shifts and by incorporation of 32 P-labelled phosphate into myelin basic protein. Results: Fibroblasts isolated from hearts of adult rats were grown in 10% serum-containing media which induced an increase in cell number by 94%. After 48 h, treatment with 10 mM NE caused an even greater increase in cell number by 222%, i.e. another 128% (comitogenic effect). In contrast, NE alone had no effect on the growth of serum-deprived cells. EGF and PDGF-AA did not replace serum as the basic mitogen. After addition of NE to proliferating cells under MAPK serum conditions, there was a rapid, time-dependent significant activation of the p42 / p44 and of CREB for up to 60 and 120 min, respectively. In both cases, the maximum of activation was reached after 5 min. Application of FO (0.1-20 mM) caused a strong MAPK activation of CREB, while no increase in the phosphorylation of the p42 / p44 was detected. Treatment with 20 mM FO led to an identical increase in cell number as application of NE. Specific blockade of PKA with RpcAMPS prevented the activation of CREB and also the comitogenic effect of FO as well as of NE. The a-and b-adrenergic receptor blocker carvedilol (10 mM) normalized all NE-induced effects. Prazosin and yohimbine, inhibitors of a -and a -adrenoceptor activation, respectively, did not influence the 1 2 NE-evoked increase in cell number. In contrast, the non-selective b-adrenoceptor blocker propranolol (1 mM) completely suppressed the comitogenic effect of NE. A similar effect was obtained with the specific b -adrenoceptor blocker ICI 118,551 (5 mM), while the 2 b -adrenoceptor blocker metoprolol did not influence the increase in cell number. Conclusions: NE elicits a comitogenic effect on 1 cultured rat cardiac fibroblasts which is prevented by b -adrenergic blockade. The activation of CREB contributes to the increase in 2 MAPK proliferation. The p42 / p44 which was also found to be activated by NE might as well be involved in the regulation of the comitogenic effect.

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Section: Introductionmentioning

confidence: 99%

Comitogenic effect of catecholamines on rat cardiac fibroblasts in culture

Leicht

Greipel

Zimmer

2000

Cardiovascular Research

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“…2). This may reflect a difference in permeability of the cells to protons in the two types of preparation; indeed, an increased membrane permeability has been reported for freshly isolated myocytes in suspension (Claycomb et al, 1984). It is also possible that the local pH near a surface (bearing cultured myocytes) may be different from that in the bulk solution, i.e.…”

Section: Measurement Of Atp Synthase Capacity Of Myocyte Mitochondriamentioning

confidence: 97%

Regulation of the mitochondrial ATP synthase in intact rat cardiomyocytes

Harris

1990

Biochemical Journal

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The ATP synthase capacity of rat heart myocytes can be measured in sonicated cell suspensions and in sonicated preparations of cultured cardiomyocytes. This procedure allows the rapid measurement of mitochondrial function in response to changes in the metabolic status of the cell. In cultured myocytes, transitions in ATP synthase capacity (with no detectable change in cellular ATP concentration) accompany a change to anoxia or electrically stimulated contraction (rise of 70%). These changes are reversed on returning to the original conditions. Exposure of myocytes to low pH has little effect on basal ATP synthase capacity (down to values less than pH 6), but markedly affects cellular ATP levels and the response of the cells to anoxia and reoxygenation, possibly mimicking changes seen in ischaemic heart. Similar effects are seen in suspensions of freshly prepared myocytes, but these preparations are less stable and more pH-sensitive than are cells in culture. It is proposed that mitochondria in vivo are directly regulated at the level of the ATP synthase, and that a regulator protein, the naturally occurring inhibitor protein from mitochondria, may be responsible for this regulation.

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“…Insulin stimulates sugar transport about l O-fold (45) and also increases incorporation of glucose into glycogen (15). Betaadrenergic stimulation enhances contractility (7), stimulates Ca 2+ currents (6,118), activates phosphorylase (28,46), adenyl cyclase (17,28,83), lipolysis (69), and protein phosphorylation (86). Alpha agonists stimulate phosphodiesterase activity (12).…”

Section: Heterogeneity a Desirable Quality For An Isolatedmentioning

confidence: 99%

Use of Isolated Adult Myocytes to Evaluate Cardiotoxicity. II. Preparation and Properties*

Haworth

1990

Toxicol Pathol

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The preparation and properties of isolated adult cardiac myocytes are reviewed, with the goal being to evaluate their usefulness as a model system for measuring cardiotoxicity. Some important factors in cell isolation methodology which impact on the quality of the preparation are identified, along with criteria for assessing the quality of cells after isolation. By all criteria, myocytes isolated by good procedures appear to largely retain their original properties. Moreover, the distinctive behavior of adult myocytes under metabolic stress endows them with a particular usefulness as monitors of toxicity. Overall, we conclude that the art of adult heart cell isolation and culture is now sufficiently advanced for either freshly isolated cells in suspension or cells in culture to be a useful model system for toxicity studies.

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Culture of the terminally differentiated ventricular cardiac muscle cell

Cited by 19 publications

References 19 publications

Comitogenic effect of catecholamines on rat cardiac fibroblasts in culture

Comitogenic effect of catecholamines on rat cardiac fibroblasts in culture

Regulation of the mitochondrial ATP synthase in intact rat cardiomyocytes

Use of Isolated Adult Myocytes to Evaluate Cardiotoxicity. II. Preparation and Properties*

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