McPherson M A, Dormer R L
Biochem J. 1984 Dec 1;224(2):473-81. doi: 10.1042/bj2240473.
A method is described for preparing isolated rat submandibular acini by collagenase digestion followed by mechanical dispersion. As assessed by Trypan Blue exclusion, phase contrast microscopy, ATP content and release of mucins and lactate dehydrogenase, the acini are morphologically and functionally intact. Secretory function of isolated acini was similar to that of intact tissue in terms of time-course, dose dependence and degree of stimulation of mucin release by adrenergic secretagogues. Mucin release was increased to the same extent (approx. 3-4-fold) by either isoproterenol or noradrenaline at a maximally effective concentration (10 microM). Stimulation of mucin release by isoproterenol (10 microM), noradrenaline (10 microM) or adrenaline (10 microM) was inhibited by propranolol (30 microM) but not by phentolamine (30 microM). Isoproterenol (10 microM) increased both 45Ca2+ uptake and efflux from the acini, which was shown to represent a net release of calcium. However, there was a delay (approx. 10 min) in onset of stimulation of 45Ca2+ mobilization which was not apparent in isoproterenol stimulation of mucin release. Our results indicate that increases in intracellular calcium mobilization in response to a beta-adrenergic secretagogue do not trigger mucin secretion from rat submandibular acini.
本文描述了一种通过胶原酶消化并随后进行机械分散来制备分离的大鼠下颌下腺腺泡的方法。通过台盼蓝排斥试验、相差显微镜检查、ATP含量以及粘蛋白和乳酸脱氢酶的释放来评估,腺泡在形态和功能上均保持完整。就时间进程、剂量依赖性以及肾上腺素能促分泌剂对粘蛋白释放的刺激程度而言,分离腺泡的分泌功能与完整组织相似。在最大有效浓度(10微摩尔)下,异丙肾上腺素或去甲肾上腺素均可使粘蛋白释放增加至相同程度(约3 - 4倍)。普萘洛尔(30微摩尔)可抑制异丙肾上腺素(10微摩尔)、去甲肾上腺素(10微摩尔)或肾上腺素(10微摩尔)对粘蛋白释放的刺激,但酚妥拉明(30微摩尔)则无此作用。异丙肾上腺素(10微摩尔)可增加腺泡对45Ca2+的摄取和外流,这表明存在钙的净释放。然而,45Ca2+动员刺激的起始存在延迟(约10分钟),而异丙肾上腺素刺激粘蛋白释放时并未出现这种延迟。我们的结果表明,β - 肾上腺素能促分泌剂引起的细胞内钙动员增加并不会触发大鼠下颌下腺腺泡的粘蛋白分泌。