Models of Cardiac Development: Transplants, Organ Culture, Cell Dispersion, and Cell Culture

Robinson, Richard B.

doi:10.1007/978-1-4613-3834-5_5

Cited by 5 publications

(3 citation statements)

References 119 publications

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“…In contrast, pure muscle cultures fail to respond to tyramine or to show increases in catecholamine levels; dopamine levels decreased significantly by days 4 and 5. Further evidence of functional Contact between nerve and muscle cells in these cultures ha § been demonstrated previously, both electrophysiologically and ultrastructurally (Robinson, 1985).…”

Section: Discussionsupporting

confidence: 75%

Neuronal regulation of the development of the alpha-adrenergic chronotropic response in the rat heart.

Drugge¹,

Rosen²,

Robinson³

1985

Circulation Research

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During development, there are changes in the response of automatic cardiac fibers to alpha-adrenergic agonists. In neonatal rat ventricle, in vitro phenylephrine (1 X 10(-8) M) induces an increase in automatic rate from 115 +/- 12 (mean +/- SEM) to 168 +/- 10 beats/min, P less than 0.05. In contrast, in adult rat ventricle, the rate decreases from 36 +/- 8 to 12 +/- 12 beats/min, P less than 0.05. At both ages, the response is attenuated by the alpha 1-antagonist, prazosin (1 X 10(-6) M). We used cultures of neonatal rat myocytes to determine whether maturation of innervation contributes to the ontogeny of this response. All non-innervated cultures showed a positive chronotropic response to alpha-stimulation; phenylephrine (1 X 10(-8) M) increased the rate from 40 +/- 2 to 52 +/- 2 beats/min, P less than 0.05. In contrast, 60% of the myocytes innervated with sympathetic neurons showed a decrease in rate in response to phenylephrine, from 78 +/- 6 to 67 +/- 6 beats/min, P less than 0.05. The negative chronotropic effect of phenylephrine did not result from the release of acetylcholine or adenosine, or the inhibition of presynaptic norepinephrine release by phenylephrine. Furthermore, exposure to neuronal norepinephrine is not responsible for the alteration in muscle cell responsiveness. In conclusion, we have demonstrated the modulation of the myocardial response to alpha-adrenergic stimulation by the occurrence of innervation in tissue culture. This provides an explanation for the previously identified ontogenetic change in alpha-adrenergic effects on intact cardiac fibers from excitation to inhibition.

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Section: Discussionsupporting

confidence: 75%

Neuronal regulation of the development of the alpha-adrenergic chronotropic response in the rat heart.

Drugge¹,

Rosen²,

Robinson³

1985

Circulation Research

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“…(Circulation Research 1989;64:1051-1062 T he entire myocardium receives postganglionic neural fibers from both the cholinergic and adrenergic nervous systems. 12 Cardiac denervation experiments in adult animals suggest that the myocardial cell membrane is dependent on this neural input for stable function. 3 Observations of human cardiac transplantation support this hypothesis.…”

mentioning

confidence: 99%

“…The study of isolated myocardial cells with and without homogeneous neurotransmission should lessen such inherent difficulties and facilitate assessment of neurotrophic modulation. 12 Cultures of isolated ventricular muscle and sinoatrial node cells, which are morphologically and functionally comparable to their in vivo source, can be prepared.…”

mentioning

confidence: 99%

Chronotropic responsiveness of developing sinoatrial and ventricular rat myocytes to autonomic agonists following adrenergic and cholinergic innervation in vitro.

Atkins

Marvin

1989

Circulation Research

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The chronotropic responses of isolated sinoatrial node and ventricular muscle cells to neurotransmitters were compared in vitro with and without selective adrenergic and cholinergic innervation. Explants of either thoracolumbar sympathetic ganglion or sacrococcygeal spinal cord were added to cultures of newborn rat sinus node regions or ventricular apexes harvested before the onset of autonomic innervation in vivo. Catecholamine synthesis was detected by glyoxylic acid histofluorescence. Acetylchoiine synthesis was indicated by monoclonal antibody labeling of choline acetyltransferase. After electrical or pharmacological stimulation of neurons, the chronotropic response of individual myocardial cells confirmed the presence of neuroeffector transmission; the nature of the myocyte response identified the stimulated neuron as either adrenergic or cholinergic. Chronotropic responses of all myocardial cells to norepinephrine or acetylchoiine were transcribed on a recorder coupled to a video photoconductive cell monitor. Isolated sinoatrial node cells were supersensitive to norepinephrine and acetylchoiine; thresholds were 3xlO' 16 M and 6xlO~1 5 M, respectively. These sinoatrial node cells remained sensitive to both norepinephrine and acetylchoiine after the development of innervation in vitro. Ventricular cells also were sensitive with thresholds of 3x10"" M and 6xlO" 14 M to norepinephrine and acetylchoiine, respectively. However, following in vitro innervation, ventricular cells were significantly less sensitive to norepinephrine and acetylchoiine (thresholds 3xlO~ M and 6 x 1 0 " M). These data are the first to demonstrate that neurotrophic modulation is not homogeneous throughout the myocardium and that it may be dependent on the specific myocardial cell innervated. (Circulation Research 1989;64:1051-1062

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Monolayer co-culture of rat heart cells and bovine adrenal chromaffin paraneurons

Trifaró

Tang

Novas

1990

In Vitro Cell Dev Biol

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This paper describes a method for the preparation of co-cultures of rat heart cells and bovine adrenal chromaffin paraneurons. The most suitable condition for heart cell isolation was when a combination of trypsin-DNAse I in Locke's solution was used for digestion. The best co-culture conditions were obtained when 10(6) heart cells were plated on 7- to 8-d-old adrenal chromaffin paraneuron cultures containing 0.5 x 10(6) cells per 35-mm diameter culture dishes. Measurements of DNA (heart cells and chromaffin paraneurons), monitoring of beating frequency (heart cells), and catecholamine (chromaffin paraneurons) levels and release indicated that both cell types remain viable and functional for several weeks. Heart cells started their characteristic contractile activity 24 h earlier when plated either on viable or lysed chromaffin paraneurons, an effect apparently due to faster surface adhesion of heart cells. The beating frequency of heart cells increased after treatment of co-cultures with either noradrenaline or nicotine, with the latter agent acting indirectly through the release of chromaffin paraneuron catecholamines. Propranolol produced a dose-related inhibition of the responses to either noradrenaline or nicotine, thus suggesting that the increase in myocyte's beating activity was mediated through beta-receptors. Anti-myosin and anti-dopamine-beta-hydroxylase immunostaining was used for cell type identification and for the demonstration of body-to-body and process-to-process contacts between adrenal chromaffin paraneurons and heart cells. This co-culture system will serve as a starting point of further studies directed to understand a) the influence of a cell type on the development and on the phenotypic characteristics of a second cell type and b) the interaction of cells derived from different organs and species.

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Models of Cardiac Development: Transplants, Organ Culture, Cell Dispersion, and Cell Culture

Cited by 5 publications

References 119 publications

Neuronal regulation of the development of the alpha-adrenergic chronotropic response in the rat heart.

Neuronal regulation of the development of the alpha-adrenergic chronotropic response in the rat heart.

Chronotropic responsiveness of developing sinoatrial and ventricular rat myocytes to autonomic agonists following adrenergic and cholinergic innervation in vitro.

Monolayer co-culture of rat heart cells and bovine adrenal chromaffin paraneurons

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