In situ localization of amylase mRNA and protein. An investigation of amylase gene activity in normal human parotid gland
In situ localization of amylase mRNA and protein. An investigation of amylase gene activity in normal human parotid gland
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DOI：
10.1177/35.1.2432114
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发表时间：
1987-01-01
影响因子：
3.2
通讯作者：
Hodes, M. E.
中科院分区：
文献类型：
--
作者：
Morley, Debra J.;Hodes, M. E.
用[(32)P]标记的淀粉酶基因探针进行原位杂交，研究了人唾液淀粉酶基因的分布。在人腮腺冰冻切片中，淀粉酶mRNA定位于腺泡细胞顶端。在导管细胞、骨骼肌或结缔组织中未观察到杂交。这些结果与淀粉酶免疫组织化学定位一致。对原位杂交技术进行了改进，以允许在不同固定的、石蜡包埋的腮腺中定位淀粉酶mRNA。虽然杂交信号随着所有固定剂的减少而减弱，但定位的模式与冰冻切片获得的模式是平行的。与常规福尔马林固定相比，乙醇-冰醋酸或Bouin液固定并无明显优势。这些结果对对基因表达研究感兴趣的研究人员具有重要的意义。我们已经证明，常规固定的人类组织石蜡切片可以用于特定mRNA的细胞定位。在免疫细胞化学的配合下，原位杂交为研究单个细胞中的mRNA和蛋白质表达提供了一个强有力的工具。
The distribution of human salivary amylase mRNA was studied by in situ hybridization to a [(32)P]-labeled amylase cDNA probe. Amylase mRNA was localized to the apical portion of acinar cells in frozen sections of human parotid salivary gland. No hybridization was noted in ductal cells, skeletal muscle, or in connective tissue. These results were consistent with immunohistochemical localization of amylase. The technique of in situ hybridization was modified to permit localization of amylase mRNA in variously fixed, paraffin-embedded parotid glands. Although the hybridization signal decreased with all fixatives, the pattern of localization paralleled that obtained with frozen sections. No advantage was noted in fixation with ethanol-acetic acid or Bouin solution over routine fixation with formalin. These results have important implications for researchers interested in studies of gene expression. We have demonstrated that routinely fixed paraffin blocks of human tissue can be used for cellular localization of specific mRNA. In coordination with immunocytochemistry, in situ hybridization offers a powerful tool for studies of mRNA and protein expression in individual cells.