Lundgren E, Terracio L, Allen D O, Borg T K
Department of Medical and Physiological Chemistry, Uppsala University, Sweden.
In Vitro Cell Dev Biol. 1988 Jan;24(1):28-34. doi: 10.1007/BF02623812.
Modulation of beta-adrenergic receptors and their ability to respond to beta-receptor stimulation was studied in cultures of adult and neonatal rat cardiac myocytes. The radioligand iodocyanopindolol (125I-CYP) was used to identify beta-adrenoceptors on the intact cells. 125I-CYP was found to bind to the receptors in a stereospecific and saturable manner. Freshly isolated neonatal and adult myocytes both had a receptor density of approximately 50 fmol/mg protein. The number of beta-receptors per milligram protein was similar during a 10-d culture period for adult myocytes but increased after a 5-d culture period for neonatal myocytes. Both cell types responded to beta-receptor stimulation with isoproterenol by a twofold increase in the concentration of cAMP and this response increased with time in culture. The number of receptors as well as the response to isoproterenol was similar for neonatal myocytes cultured on laminin, collagen type I, or on uncoated culture dishes. From these data we conclude that cultured cardiac myocytes maintain functional beta-receptors as they progress into culture, and the expression of beta-receptors is not influenced by culture substrates.
在成年和新生大鼠心肌细胞培养物中研究了β-肾上腺素能受体的调节及其对β-受体刺激的反应能力。放射性配体碘氰吲哚洛尔(125I-CYP)用于识别完整细胞上的β-肾上腺素能受体。发现125I-CYP以立体特异性和可饱和的方式与受体结合。新鲜分离的新生和成年心肌细胞的受体密度均约为50 fmol/mg蛋白质。在10天的培养期内,成年心肌细胞每毫克蛋白质的β-受体数量相似,但新生心肌细胞在培养5天后增加。两种细胞类型对异丙肾上腺素的β-受体刺激的反应是cAMP浓度增加两倍,并且这种反应随着培养时间的延长而增加。在层粘连蛋白、I型胶原或未包被的培养皿上培养的新生心肌细胞,其受体数量以及对异丙肾上腺素的反应相似。从这些数据我们得出结论,培养的心肌细胞在进入培养过程中保持功能性β-受体,并且β-受体的表达不受培养底物的影响。