IMPROVED TISSUE PREPARATION FOR IN SITU LOCALIZATION OF SPECIFIC mRNA USING NON-RADIOACTIVE DNA PROBES : EFFECTS OF PROTEASE DIGESTION AND PROBE SIZE ON SIGNAL DETECTION IN FROZEN AND PARAFFIN SECTIONS OF RAT PITUITARY GLANDS (<Symposium I>Molecular Cytoc
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概要
- 論文の詳細を見る
To better describe the physiological state of cells, detection of specific mRNA by in situ hybridization at the cellular level is often demanded. In order to accomplish this reliably, various factors such as morphological preservation, retention of mRNA, accessibility of the labeled probe to the target mRNA and efficiency of the hybridization should be considered during the tissue processing. In this paper, using non-radioactive probes, we investigated the effects of protease treatment and the size of probe on the eficiency of in situ hybridization with mRNAs in frozen and paraffin-embedded tissue sections of the rat pituitary glands. As non-radioactive haptenic DNA probes, we used dinitrophenyl (DNP)-labeled pro-opiomelanocortin (POMC) DNA and thymine-thymine (T-T) dimerized prolactin CDNA. The signals were visualized by the indirect enzyme-immunohistochemistry. The best results were obtained when frozen sections of tissues fixed by 4% paraformaldehyde in phosphate buffered saline were mildly digested with protcase and hybridized with thc haptenized DNAs of about 200-400 base pairs.
- 日本組織細胞化学会の論文
著者
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Koji Takehiko
Department Of Histology And Cell Biology Nagasaki University Graduate School Of Biomedical Sciences
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Nakane Paul
長崎大学 解剖3
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Moriuchi Tetsuya
Hokkaido Urothelial Cancer Research Group
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NAKANE Paul
Department of Cell Biology, School of Medicine, Tokai University
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MORIUCHI Tetsuya
Department of Cell Biology, Tokai University School of Medicine
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Moriuchi Tetsuya
Department Of Cell Biology Tokai University School Of Medicine
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Koji Takehiko
Department Fo Histology And Cell Biology Nagasaki University School Of Medicine
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NAKANE Paul
Environmental Biotechnology Institute, California Polytechnic State University
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Nakane Paul
Department Of Anatomy Nagasaki University School Of Medicine
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Mochizuki T
Shizuoka Cancer Center Hospital And Res. Inst. Shizuoka Jpn
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Nakane P
Essential Therapeutics Ca Usa
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Nakane Paul
Department Of Anatomy (iii) Nagasaki University School Of Medicine
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