Dvorak A M, Morgan E S, Lichtenstein L M, Weller P F, Schleimer R P
Department of Pathology, Harvard Medical School and Beth Israel Deaconess Medical Center, Boston, Massachusetts 02215, USA.
J Histochem Cytochem. 2000 Jan;48(1):1-12. doi: 10.1177/002215540004800101.
The distribution of ribosomes in mature human mast cells, a major granulated secretory cell, does not resemble that in other secretory cells, such as pancreatic acinar cells and plasma cells. By routine ultrastructural analysis, ribosomes in human mast cells are often close to, attached to, or even appear to be within secretory granules. To document better these relationships, we used multiple electron microscopic imaging methods, based on different principles, to define RNA, ribosome, and granule relationships in mature human mast cells. These methods included EDTA regressive staining, RNase digestion, immunogold labeling of ribonucleoproteins or uridine, direct binding or binding after ultrastructural in situ hybridization of various polyuridine probes to polyadenine in mRNA, and ultrastructural autoradiographic localization of [3H]-uridine incorporated into cultured human mast cells. These different labeling methods demonstrated ribosomes, RNA, U1SnRNP (a small nuclear RNP specific for alternative splicing of mRNA), mRNA, and uridine to be associated with secretory granules in human mast cells, implicating granules in a larger synthetic role in mast cell biology.
成熟人类肥大细胞是一种主要的颗粒分泌细胞,其核糖体的分布与其他分泌细胞不同,如胰腺腺泡细胞和浆细胞。通过常规超微结构分析,人类肥大细胞中的核糖体常常靠近、附着于分泌颗粒,甚至似乎位于分泌颗粒内部。为了更好地记录这些关系,我们基于不同原理使用了多种电子显微镜成像方法,以确定成熟人类肥大细胞中RNA、核糖体和颗粒之间的关系。这些方法包括乙二胺四乙酸(EDTA)递减染色、核糖核酸酶消化、核糖核蛋白或尿苷的免疫金标记、各种聚尿苷探针与mRNA中的聚腺苷酸进行超微结构原位杂交后的直接结合或结合,以及对掺入培养的人类肥大细胞中的[3H] - 尿苷进行超微结构放射自显影定位。这些不同的标记方法表明,核糖体、RNA、U1SnRNP(一种对mRNA可变剪接具有特异性的小核核糖核蛋白)、mRNA和尿苷与人类肥大细胞中的分泌颗粒相关,这意味着颗粒在肥大细胞生物学中具有更大的合成作用。