Molecules 2011, 16(5), 3618-3635; doi:10.3390/molecules16053618
Review

Molecular Morphology of Pituitary Cells, from Conventional Immunohistochemistry to Fluorescein Imaging

Akira Matsuno 1,* email, Akiko Mizutani 1,2 email, Hiroko Okinaga 2 email, Koji Takano 3 email, So Yamada 1 email, Shoko M. Yamada 1 email, Hiroshi Nakaguchi 1 email, Katsumi Hoya 1 email, Mineko Murakami 1 email, Masato Takeuchi 4 email, Mutsumi Sugaya 4 email, Johbu Itoh 5 email, Susumu Takekoshi 6 email and R. Yoshiyuki Osamura 7 email
1 Department of Neurosurgery, Teikyo University Chiba Medical Center, Chiba 299-0111, Japan 2 Teikyo Heisei University, Tokyo 170-8445, Japan 3 Department of Nephrology and Endocrinology, University of Tokyo Hospital, Tokyo 113-0033, Japan 4 Department of Rehabilitation, Teikyo University Chiba Medical Center, Chiba 299-0111, Japan 5 Teaching and Research Support Center, Tokai University School of Medicine, Kanagawa 259-1100, Japan 6 Department of Pathology, Tokai University School of Medicine, Kanagawa 259-1100, Japan 7 Pathology Diagnosis Center, International University of Health and Welfare, Tokyo 108-8329, Japan
* Author to whom correspondence should be addressed.
Received: 9 March 2011; in revised form: 25 April 2011 / Accepted: 26 April 2011 / Published: 29 April 2011
(This article belongs to the Special Issue Fluorophores - The Fluorescent Toolbox in Biological and Biomedical Research)
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Abstract: In situ hybridization (ISH) at the electron microscopic (EM) level is essential for elucidating the intracellular distribution and role of mRNA in protein synthesis. EM-ISH is considered to be an important tool for clarifying the intracellular localization of mRNA and the exact site of pituitary hormone synthesis on the rough endoplasmic reticulum. A combined ISH and immunohistochemistry (IHC) under EM (EM-ISH&IHC) approach has sufficient ultrastructural resolution, and provides two-dimensional images of the subcellular localization of pituitary hormone and its mRNA in a pituitary cell. The advantages of semiconductor nanocrystals (quantum dots, Qdots) and confocal laser scanning microscopy (CLSM) enable us to obtain three-dimensional images of the subcellular localization of pituitary hormone and its mRNA. Both EM-ISH&IHC and ISH & IHC using Qdots and CLSM are useful for understanding the relationships between protein and mRNA simultaneously in two or three dimensions. CLSM observation of rab3B and SNARE proteins such as SNAP-25 and syntaxin has revealed that both rab3B and SNARE system proteins play important roles and work together as the exocytotic machinery in anterior pituitary cells. Another important issue is the intracellular transport and secretion of pituitary hormone. We have developed an experimental pituitary cell line, GH3 cell, which has growth hormone (GH) linked to enhanced yellow fluorescein protein (EYFP). This stable GH3 cell secretes GH linked to EYFP upon stimulation by Ca2+ influx or Ca2+ release from storage. This GH3 cell line is useful for the real-time visualization of the intracellular transport and secretion of GH. These three methods from conventional immunohistochemistry and fluorescein imaging allow us to consecutively visualize the process of transcription, translation, transport and secretion of anterior pituitary hormone.
Keywords: pituitary hormone; mRNA; intracellular transport and secretion; quantum dot; enhanced yellow fluorescein protein

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Cite This Article

MDPI and ACS Style

Matsuno, A.; Mizutani, A.; Okinaga, H.; Takano, K.; Yamada, S.; Yamada, S.M.; Nakaguchi, H.; Hoya, K.; Murakami, M.; Takeuchi, M.; Sugaya, M.; Itoh, J.; Takekoshi, S.; Osamura, R.Y. Molecular Morphology of Pituitary Cells, from Conventional Immunohistochemistry to Fluorescein Imaging. Molecules 2011, 16, 3618-3635.

AMA Style

Matsuno A, Mizutani A, Okinaga H, Takano K, Yamada S, Yamada SM, Nakaguchi H, Hoya K, Murakami M, Takeuchi M, Sugaya M, Itoh J, Takekoshi S, Osamura RY. Molecular Morphology of Pituitary Cells, from Conventional Immunohistochemistry to Fluorescein Imaging. Molecules. 2011; 16(5):3618-3635.

Chicago/Turabian Style

Matsuno, Akira; Mizutani, Akiko; Okinaga, Hiroko; Takano, Koji; Yamada, So; Yamada, Shoko M.; Nakaguchi, Hiroshi; Hoya, Katsumi; Murakami, Mineko; Takeuchi, Masato; Sugaya, Mutsumi; Itoh, Johbu; Takekoshi, Susumu; Osamura, R. Yoshiyuki. 2011. "Molecular Morphology of Pituitary Cells, from Conventional Immunohistochemistry to Fluorescein Imaging." Molecules 16, no. 5: 3618-3635.

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