Krall J F, Morin A
J Cell Physiol. 1986 Nov;129(2):250-6. doi: 10.1002/jcp.1041290217.
Cells growing in culture with previously described properties of rat uterine smooth muscle accumulated 45Ca2+ from the medium. Ca2+ uptake by these cells was stimulated by the addition to the medium of 8-bromo-cGMP but not by 8-bromo-cAMP. Ca2+ uptake was also stimulated by carbachol and by the nitro-vasodilator nitroprusside. Although cholinergic agonists have been shown previously to stimulate contraction but not cGMP synthesis in the rat myometrium, both carbachol and nitroprusside stimulated cGMP production by the cultured cells. These results suggested the cells had cholinergic receptor-mediated functions that reflected some neurotransmitter-sensitive properties of uterine smooth muscle in situ. When determined by a specific radioligand binding assay, subcellular fractions of the cultured cells bound muscarinic cholinergic agonists and antagonists with affinities expected of the muscarinic receptor. The cells were also sensitive to the beta-adrenergic catecholamine agonist isoproterenol, which stimulated cAMP production but not Ca2+ uptake. Carbachol failed to inhibit isoproterenol-dependent cAMP production, which is an important property of the cholinergic receptor in uterine smooth muscle in situ. These results suggest some but not all acetylcholine-sensitive properties of uterine smooth muscle may be retained in cell culture.
在培养中生长的具有先前所述大鼠子宫平滑肌特性的细胞从培养基中积累了45Ca2+。向培养基中添加8-溴-cGMP可刺激这些细胞摄取Ca2+,但添加8-溴-cAMP则无此作用。卡巴胆碱和硝基血管扩张剂硝普钠也可刺激Ca2+摄取。尽管先前已表明胆碱能激动剂可刺激大鼠子宫肌层收缩,但不刺激cGMP合成,但卡巴胆碱和硝普钠均可刺激培养细胞产生cGMP。这些结果表明,这些细胞具有胆碱能受体介导的功能,反映了子宫平滑肌在原位的一些神经递质敏感特性。通过特异性放射性配体结合试验测定时,培养细胞的亚细胞组分以毒蕈碱受体预期的亲和力结合毒蕈碱胆碱能激动剂和拮抗剂。这些细胞对β-肾上腺素能儿茶酚胺激动剂异丙肾上腺素也敏感,异丙肾上腺素可刺激cAMP产生,但不刺激Ca2+摄取。卡巴胆碱未能抑制异丙肾上腺素依赖性cAMP产生,这是子宫平滑肌原位胆碱能受体的一个重要特性。这些结果表明,子宫平滑肌的一些但并非全部乙酰胆碱敏感特性可能在细胞培养中得以保留。