An immunocytochemical technique for analysis of regulation of genes encoding early differentiation marker antigens in an oocyte translation system.
Monoclonal antibodies are useful probes for analyzing cells at the molecular level at various developmental stages. Although identification of the genes encoding tissue- and stage-specific antigens could be informative for further molecular analysis, gene cloning is usually a time-consuming step, particularly when a monoclonal antibody is the only probe available. We describe here an immunocytochemical method for preliminary and immediate analysis of the regulation of antigen-coding genes. mRNAs purified from stage 27 and 38 Xenopus tadpoles were fractionated by size and injected into newt oocytes, from which frozen sections were prepared for immunostaining with tissue-specific monoclonal antibodies. Both of the antigens we tested, which are early markers for differentiating epidermal cells of Xenopus tadpoles, were detected in mRNA injected oocytes, but not in control oocytes. Immunostaining for each of the antigens showed that their relative levels in stage 27 and 38 tadpole tissue were reflected in those oocytes injected with mRNA purified from tadpoles of the respective stages. We suggest that this oocyte translation system combined with immunostaining provides for rapid analysis of changes in levels of antigen coding mRNAs throughout development.
Nishizawa
K
K
Department of Neurobiology, Institute for Brain Research, Tokyo University School of Medicine, Japan.
Okado
H
H
Okamoto
H
H
D000818:Animals; D000911:Antibodies, Monoclonal; D000943:Antigens, Differentiation / Q000096:biosynthesis* / Q000235:genetics; D001001:Anura; D005260:Female; D005786:Gene Expression Regulation; D007150:Immunohistochemistry / Q000379:methods*; D008297:Male; D008845:Microinjections; D009865:Oocytes / Q000166:cytology / Q000378:metabolism*; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000008:administration & dosage* / Q000302:isolation & purification / Q000378:metabolism*; D012455:Salamandridae; D014981:Xenopus
D000911:Antibodies, Monoclonal; D000943:Antigens, Differentiation; D012333:RNA, Messenger