Reggio H A, Palade G E
J Cell Biol. 1978 May;77(2):288-314. doi: 10.1083/jcb.77.2.288.
Sulfate incorporation into the guinea pig pancreas was investigated by light (LM) and electron microscope (EM) autoradiography using a system of minilobules incubated in vitro for 60 min in Krebs-Ringer bicarbonate medium (KRB) containing 35SO4(-2). In acinar cells, examined by EM autoradiography, the label was found concentrated over Golgi elements (including condensing vacuoles) and zymogen granules. 35SO4(-2) was also incorporated by the epithelial cells of the entire pancreatic duct system, the incorporation being surprisingly high in the epithelium of the major ducts. In all ductal epithelia, autoradiographic grains appeared over the Golgi complex and the plasmalemma. Since a contribution of duct epithelium to the sulfated compounds found in the discharged secretion could not be ruled out, a purified zymogen granule fraction was used as a source material for the isolation of sulfated compounds of acinar origin. The presence of 35S-radioactivity in the zymogen granules and condensing vacuoles of this fraction was ascertained by autoradiography (of sectioned pellets). From a lysate of this zymogen granule fraction, a soluble sulfated compound of low isoelectric point and high molecular weight was isolated by gel filtration under conditions that allowed its satisfactory separation from the bulk of the secretory proteins.
利用在含有35SO4(-2)的Krebs-林格碳酸氢盐培养基(KRB)中体外孵育60分钟的小胰小叶系统,通过光学显微镜(LM)和电子显微镜(EM)放射自显影术研究了硫酸盐掺入豚鼠胰腺的情况。在通过EM放射自显影术检查的腺泡细胞中,发现标记物集中在高尔基体成分(包括浓缩泡)和酶原颗粒上。整个胰腺导管系统的上皮细胞也掺入了35SO4(-2),在主导管的上皮细胞中掺入量惊人地高。在所有导管上皮中,放射自显影颗粒出现在高尔基体复合体和质膜上。由于不能排除导管上皮对排出分泌物中发现的硫酸化化合物的贡献,因此使用纯化的酶原颗粒部分作为分离腺泡来源的硫酸化化合物的原料。通过(切片沉淀的)放射自显影术确定了该部分的酶原颗粒和浓缩泡中存在35S放射性。从该酶原颗粒部分的裂解物中,在允许其与大部分分泌蛋白满意分离的条件下,通过凝胶过滤分离出一种低等电点和高分子量的可溶性硫酸化化合物。