{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,31]],"date-time":"2025-12-31T17:53:14Z","timestamp":1767203594922,"version":"build-2238731810"},"reference-count":79,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2011,4,29]],"date-time":"2011-04-29T00:00:00Z","timestamp":1304035200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/3.0\/"}],"content-domain":{"domain":["www.mdpi.com"],"crossmark-restriction":true},"short-container-title":["Molecules"],"abstract":"<jats:p>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&amp;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&amp;IHC and ISH &amp; 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.<\/jats:p>","DOI":"10.3390\/molecules16053618","type":"journal-article","created":{"date-parts":[[2011,5,4]],"date-time":"2011-05-04T09:03:57Z","timestamp":1304499837000},"page":"3618-3635","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Molecular Morphology of Pituitary Cells, from Conventional Immunohistochemistry to Fluorescein Imaging"],"prefix":"10.3390","volume":"16","author":[{"given":"Akira","family":"Matsuno","sequence":"first","affiliation":[{"name":"Department of Neurosurgery, Teikyo University Chiba Medical Center, Chiba 299-0111, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Akiko","family":"Mizutani","sequence":"additional","affiliation":[{"name":"Department of Neurosurgery, Teikyo University Chiba Medical Center, Chiba 299-0111, Japan"},{"name":"Teikyo Heisei University, Tokyo 170-8445, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hiroko","family":"Okinaga","sequence":"additional","affiliation":[{"name":"Teikyo Heisei University, Tokyo 170-8445, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Koji","family":"Takano","sequence":"additional","affiliation":[{"name":"Department of Nephrology and Endocrinology, University of Tokyo Hospital, Tokyo 113-0033, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"So","family":"Yamada","sequence":"additional","affiliation":[{"name":"Department of Neurosurgery, Teikyo University Chiba Medical Center, Chiba 299-0111, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shoko M.","family":"Yamada","sequence":"additional","affiliation":[{"name":"Department of Neurosurgery, Teikyo University Chiba Medical Center, Chiba 299-0111, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hiroshi","family":"Nakaguchi","sequence":"additional","affiliation":[{"name":"Department of Neurosurgery, Teikyo University Chiba Medical Center, Chiba 299-0111, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Katsumi","family":"Hoya","sequence":"additional","affiliation":[{"name":"Department of Neurosurgery, Teikyo University Chiba Medical Center, Chiba 299-0111, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mineko","family":"Murakami","sequence":"additional","affiliation":[{"name":"Department of Neurosurgery, Teikyo University Chiba Medical Center, Chiba 299-0111, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Masato","family":"Takeuchi","sequence":"additional","affiliation":[{"name":"Department of Rehabilitation, Teikyo University Chiba Medical Center, Chiba 299-0111, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mutsumi","family":"Sugaya","sequence":"additional","affiliation":[{"name":"Department of Rehabilitation, Teikyo University Chiba Medical Center, Chiba 299-0111, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Johbu","family":"Itoh","sequence":"additional","affiliation":[{"name":"Teaching and Research Support Center, Tokai University School of Medicine, Kanagawa 259-1100, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Susumu","family":"Takekoshi","sequence":"additional","affiliation":[{"name":"Department of Pathology, Tokai University School of Medicine, Kanagawa 259-1100, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"R. Yoshiyuki","family":"Osamura","sequence":"additional","affiliation":[{"name":"Pathology Diagnosis Center, International University of Health and Welfare, Tokyo 108-8329, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2011,4,29]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"265","DOI":"10.1007\/BF00269162","article-title":"Ultrastructural distribution of growth hormone, prolactin mRNA in normal rat pituitary cells: A comparison between preembedding and postembedding methods","volume":"102","author":"Matsuno","year":"1994","journal-title":"Histochemistry"},{"key":"ref_2","first-page":"771","article-title":"Application of biotinylated oligonucleotide probes to the detection of pituitary hormone mRNA using Northern blot analysis, in situ hybridization at light and electron microscopic levels","volume":"26","author":"Matsuno","year":"1994","journal-title":"Histochem. 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