<I>In situ</I> RT-PCR: Identification of specific cells using a high sensitive detection of mRNA

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  • NAKAKURA Takashi
    Integrated Bioscience Section, Graduate School of Science and Technology, Shizuoka University
  • TANAKA Shigeyasu
    Integrated Bioscience Section, Graduate School of Science and Technology, Shizuoka University

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Other Title
  • 高感度ｍＲＮＡ検出法で細胞を同定する―ｉｎ　ｓｉｔｕ　ＲＴ‐ＰＣＲ法―
  • 高感度mRNA検出法で細胞を同定する
  • コウカンド mRNA ケンシュツホウ デ サイボウ オ ドウテイスル

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Abstract

Recently, in situ reverse transcription (RT)-PCR has been developed to detect weak mRNA signals on histological sections by amplifying the relevant molecular mRNA, because RT-PCR allows one mRNA to be amplified a billionfold. This method was performed using rTth DNA polymerase that transcripts cDNA to mRNA, and amplifying the cDNA. We applied this technique to detect prohormone convertases PC1 and PC2 in the rat pituitary gland, and vascular endothelial growth factor (VEGF) in the developing pituitary gland, where these mRNAs were not detectable usual in situ hybridization techniques. Therefore, this in situ RT-PCR method is useful for identifying specific cells expressing weak signals in histological sections.

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