Bendayan M, Orstavik T B
J Histochem Cytochem. 1982 Jan;30(1):58-66. doi: 10.1177/30.1.6915073.
The subcellular localization of kallikrein was studied in the rat pancreas using the immunocytochemical protein A-gold technique. Kallikrein was found at the level of the rough endoplasmic reticulum (RER), Golgi cisternae, condensing vacuoles, and zymogen granules of the pancreatic acinar cells as well as in the acinar lumen. The effect of various tissue processings on the immunocytochemical labeling of kallikrein was evaluated using pancreatic tissue fixed in glutaraldehyde and embedded in Epon, Lowicryl K4M, or glycol methacrylate (GMA). Compared to the results obtained with Epon, Lowicryl allowed improved resolution and specificity in the immunocytochemical labeling, while GMA retained greater amounts of kallikrein antigenicity leading to a higher intensity in the labeling; since it also gave a good ultrastructural preservation, GMA appeared to be the superior embedding medium for the localization of kallikrein. The quantitative evaluation of the labeling obtained under the three embedding conditions showed the presence of an increasing concentration gradient along the RER-Golgi-granule secretory pathway, suggesting that, like other pancreatic exocrine enzymes, kallikrein is synthesized in the RER, processed through the Golgi apparatus, and packed in the zymogen granules before being released into the acinar lumen.
利用免疫细胞化学蛋白A-金技术研究了大鼠胰腺中激肽释放酶的亚细胞定位。在胰腺腺泡细胞的粗面内质网(RER)、高尔基池、浓缩泡和酶原颗粒水平以及腺泡腔内发现了激肽释放酶。使用戊二醛固定并包埋在环氧树脂、Lowicryl K4M或甲基丙烯酸乙二醇酯(GMA)中的胰腺组织,评估了各种组织处理对激肽释放酶免疫细胞化学标记的影响。与环氧树脂获得的结果相比,Lowicryl在免疫细胞化学标记中具有更高的分辨率和特异性,而GMA保留了更多的激肽释放酶抗原性,导致标记强度更高;由于它也能很好地保存超微结构,GMA似乎是用于激肽释放酶定位的最佳包埋介质。在三种包埋条件下获得的标记的定量评估显示,沿着RER-高尔基体-颗粒分泌途径存在浓度梯度增加,这表明,与其他胰腺外分泌酶一样,激肽释放酶在RER中合成,通过高尔基体加工,并在释放到腺泡腔之前包装在酶原颗粒中。