In situ localization of amylase mRNA and protein. An investigation of amylase gene activity in normal human parotid gland.
The distribution of human salivary amylase mRNA was studied by in situ hybridization to a [32P]-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.
D000681:Amylases / Q000235:genetics*; D005796:Genes; D006652:Histological Techniques; D006801:Humans; D009693:Nucleic Acid Hybridization; D010306:Parotid Gland / Q000032:analysis* / Q000201:enzymology; D011506:Proteins / Q000032:analysis*; D012333:RNA, Messenger / Q000032:analysis*; D013997:Time Factors
D011506:Proteins; D012333:RNA, Messenger; D000681:Amylases