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果蝇乙醇脱氢酶基因的组织特异性表达:原位杂交与免疫细胞化学的比较

Tissue specific expression of the Drosophila Adh gene: a comparison of in situ hybridization and immunocytochemistry.

作者信息

Anderson S M, Brown M R, McDonald J F

机构信息

Department of Biology, UNC, Greensboro 27412.

出版信息

Genetica. 1991;84(2):95-100. doi: 10.1007/BF00116548.

Abstract

The tissue specific patterns for Drosophila melanogaster alcohol dehydrogenase (Adh) mRNA and protein expression were determined using in situ hybridization and immunocytochemical techniques. Alcohol dehydrogenase mRNAs were localized in thin sections of frozen tissue using the hybridization of single stranded RNA probes. Alcohol dehydrogenase protein was identified in frozen tissue samples using ADH antisera, a biotinylated secondary antibody, and streptavidin conjugated to horseradish peroxidase. In tissues such as fat body, gastric caeca, and adult cardiac valve, the patterns of expression for ADH protein and mRNA were identical. Other tissues such as oocytes, nurse cells, imaginal disks, and brain show levels of Adh mRNA that are lower than or equivalent to those observed in the previously mentioned tissues, but they exhibit little or no ADH protein. The lack of concordance between Adh mRNA and ADH protein expression in oocytes and nurse cells may reflect the packaging of maternal mRNAs (but not ADH protein) for use in early development. The reason(s) for the other discrepancies in protein and mRNA expression are not known at this time but may be due to post-transcriptional regulation in these specific tissues.

摘要

利用原位杂交和免疫细胞化学技术确定了黑腹果蝇乙醇脱氢酶(Adh)mRNA和蛋白质表达的组织特异性模式。使用单链RNA探针杂交在冷冻组织的薄片中定位乙醇脱氢酶mRNA。使用ADH抗血清、生物素化二抗以及与辣根过氧化物酶偶联的链霉亲和素在冷冻组织样本中鉴定乙醇脱氢酶蛋白。在脂肪体、胃盲囊和成年心脏瓣膜等组织中,ADH蛋白和mRNA的表达模式是相同的。其他组织如卵母细胞、滋养细胞、成虫盘和脑,其Adh mRNA水平低于或等同于上述组织中观察到的水平,但它们几乎没有或没有ADH蛋白。卵母细胞和滋养细胞中Adh mRNA与ADH蛋白表达缺乏一致性可能反映了母源mRNA(而非ADH蛋白)用于早期发育的包装情况。目前尚不清楚蛋白质和mRNA表达中其他差异的原因,但可能是由于这些特定组织中的转录后调控。

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