title: Functional molecular morphology of anterior pituitary cells, from hormone production to intracellular transport and secretion.
abstract: Combined in situ hybridization (ISH) and immunohistochemistry (IHC) under electron microscopy (EM-ISH & IHC) has sufficient ultrastructural resolution to provide two-dimensional images of 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 relationship between protein and mRNA simultaneously in two or three dimensions. CLSM observation of rab3B and SNARE proteins such as SNAP-25 and syntaxin revealed that both rab3B and SNARE system proteins play an important role and work together as the exocytotic machinery in anterior pituitary cells. Another important issue is the intracellular transport and secretion of pituitary hormone. An experimental pituitary cell line, the GH3 cell, in which growth hormone (GH) is linked to enhanced yellow fluorescein protein (EYFP), has been developed. This stable GH3 cell secretes GH linked to EYFP upon being stimulated by Ca(2+) influx or Ca(2+) release from storage. This GH3 cell is useful for real-time visualization of the intracellular transport and secretion of GH. These three methods enable us to visualize consecutively the processes of transcription, translation, transport, and secretion of pituitary hormone.
authors: Department of Neurosurgery, Teikyo University Chiba Medical Center, 3426-3 Anesaki, Ichihara, Chiba, 299-0111, Japan. akirakun@med.teikyo-u.ac.jp
authors: Akira
authors: A
authors: Matsuno
authors: Akiko
authors: A
authors: Mizutani
authors: Hiroko
authors: H
authors: Okinaga
authors: Koji
authors: K
authors: Takano
authors: So
authors: S
authors: Yamada
authors: Shoko M
authors: SM
authors: Yamada
authors: Hiroshi
authors: H
authors: Nakaguchi
authors: Katsumi
authors: K
authors: Hoya
authors: Mineko
authors: M
authors: Murakami
authors: Masato
authors: M
authors: Takeuchi
authors: Mutsumi
authors: M
authors: Sugaya
authors: Johbu
authors: J
authors: Itoh
authors: Susumu
authors: S
authors: Takekoshi
authors: R Yoshiyuki
authors: RY
authors: Osamura
mesh_terms: D000818:Animals; D001426:Bacterial Proteins; D001692:Biological Transport / Q000502:physiology; D002460:Cell Line; D005089:Exocytosis / Q000502:physiology; D013006:Growth Hormone / Q000378:metabolism; D006801:Humans; D021621:Imaging, Three-Dimensional / Q000379:methods*; D007150:Immunohistochemistry / Q000379:methods; D017403:In Situ Hybridization / Q000379:methods; D008164:Luminescent Proteins; D018613:Microscopy, Confocal / Q000379:methods; D008854:Microscopy, Electron / Q000379:methods; D010902:Pituitary Gland / Q000378:metabolism / Q000648:ultrastructure; D050765:Qa-SNARE Proteins / Q000378:metabolism / Q000648:ultrastructure; D045663:Quantum Dots; D012333:RNA, Messenger / Q000378:metabolism*; D051381:Rats; D050825:Synaptosomal-Associated Protein 25 / Q000378:metabolism / Q000648:ultrastructure; D020692:rab3 GTP-Binding Proteins / Q000378:metabolism / Q000648:ultrastructure
chemical_list: D001426:Bacterial Proteins; D008164:Luminescent Proteins; D050765:Qa-SNARE Proteins; D012333:RNA, Messenger; C494210:Snap25 protein, rat; D050825:Synaptosomal-Associated Protein 25; C054562:yellow fluorescent protein, Bacteria; D013006:Growth Hormone; C492132:Rab3b protein, rat; D020692:rab3 GTP-Binding Proteins