Developmental profile and tissue distribution of Drosophila alcohol dehydrogenase: an immunochemical analysis with monoclonal antibodies.
To analyze Drosophila alcohol dehydrogenase gene (Adh) expression and tissue distribution at various developmental stages, we devised several immunochemical techniques making use of monoclonal antibodies against Drosophila alcohol dehydrogenase (ADH), which had been obtained previously. We here report their application to analyze the expression of Adh in a wild-type strain of D. melanogaster. s-ELISA tests were performed to evaluate fluctuations in ADH content and specific activity during development in individual organs as well as in whole individuals. In all cases, ADH specific activity appeared to be quite constant, which implies that variations in enzyme activity reflect differences in protein content. Immunoblottings of crude homogenates revealed immunoreactive low relative molecular mass peptides in addition to the 27 KD monomeric band, showing a conserved banding pattern in different organs and developmental stages. Immunohistochemical assays on whole organs were used to analyze the general pattern of ADH distribution. Immunoperoxidase staining of cryosections proved to be of crucial relevance, as it yielded full details of the tissue localization of ADH within the ADH-positive organs. We have shown not only that ADH displays a specific distribution in some organs but also that the enzyme is restricted to certain cell types.
Visa
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Departament de Genètica, Facultat de Biologia, Universitat de Barcelona, Spain.
Fibla
J
J
Santa-Cruz
M C
MC
Gonzàlez-Duarte
R
R
D000426:Alcohol Dehydrogenase / Q000032:analysis* / Q000276:immunology; D000818:Animals; D000911:Antibodies, Monoclonal; D015153:Blotting, Western; D002457:Cell Extracts / Q000737:chemistry; D004331:Drosophila melanogaster / Q000033:anatomy & histology / Q000201:enzymology*; D005629:Frozen Sections; D015870:Gene Expression; D007124:Immunoenzyme Techniques; D007150:Immunohistochemistry / Q000379:methods*; D007527:Isoenzymes / Q000032:analysis / Q000276:immunology; D007814:Larva / Q000033:anatomy & histology / Q000201:enzymology; D014018:Tissue Distribution
D000911:Antibodies, Monoclonal; D002457:Cell Extracts; D007527:Isoenzymes; D000426:Alcohol Dehydrogenase