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Electron microscopic and confocal laser scanning microscopic observation of subcellular organelles and pituitary hormone mRNA: Application of ultrastructuralIn situ hybridization and immunohistochemistry to the pathophysiological studies of pituitary cells

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Abstract

Nonradioisotopic electron microscopic (EM)in situ hybridization (ISH) (EM-ISH) with biotinylated oligonucleotide probes is utilized for the ultrastructural visualization of pituitary hormone mRNA in rat pituitary cells. EM-ISH is an important tool for clarifying the intracellular localization of mRNA and the exact site of specific hormone synthesis on the rough endoplasmic reticulum. The simultaneous visualization of mRNA and encoded protein in the same cell using preembedding EM-ISH and subsequent postembedding immunoreaction with protein A colloidal gold complex can provide an important clue for elucidating the intracellular correlation of mRNA translation and secretion of translated protein. Another focus of this review is the utilization of a recently developed imaging system of confocal laser scanning microscopy (CLSM). The combination of CLSM and image analysis system (IAS) enables us to visualize an individual dimensional image of the intracellular distribution of mRNA and subcellular organelles successfully at any optional cross sections of light microscopic ISH studies, and can be another useful tool for the ultrastructural ISH study of mRNA.

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Authors and Affiliations

  1. Department of Neurosurgery, Teikyo University Ichihara Hospital, 3426-3 Anegasaki, 299-0111, Ichihara City, Chiba, Japan

    Akira Matsuno & Tadashi Nagashima

  2. Laboratories for Structure and Function Research, Tokai University School of Medicine, Boseidai, 259-1100, Isehara City, Kanagawa, Japan

    Johbu Itoh

  3. Department of Pathology, Tokai University School of Medicine, Boseidai, 259-1100, Isehara City, Kanagawa, Japan

    R. Yoshiyuki Osamura & Keiichi Watanabe

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  1. Akira Matsuno
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  2. Johbu Itoh
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  3. R. Yoshiyuki Osamura
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  4. Keiichi Watanabe
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  5. Tadashi Nagashima
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Matsuno, A., Itoh, J., Osamura, R.Y. et al. Electron microscopic and confocal laser scanning microscopic observation of subcellular organelles and pituitary hormone mRNA: Application of ultrastructuralIn situ hybridization and immunohistochemistry to the pathophysiological studies of pituitary cells. Endocr Pathol 10, 199–211 (1999). https://doi.org/10.1007/BF02738881

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