title: Analyses of the mechanism of intracellular transport and secretion of pituitary hormone, with an insight of the subcellular localization of pituitary hormone and its mRNA.
abstract: Combined in situ hybridization (ISH) and immunohistochemistry (IHC) under electron microscopy (EM-ISH&IHC) has sufficient ultrastructural resolution and provides 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 subcellular localization of pituitary hormone and its mRNA. Both EM-ISH&IHC and ISH&IHC using Qdots and CLSM are useful for understanding the relation between protein and mRNA simultaneously in two or three dimensions. Another important issue is the intracellular transport and secretion of pituitary hormone. We have developed an experimental pituitary cell line, the GH3 cell, which has growth hormone (GH) linked to enhanced yellow fluorescein protein (EYFP). This stable GH3 cell secretes GH linked to EYFP upon stimulated by Ca2+ influx or Ca2+ 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 process of transcription, translation, transport, and secretion of pituitary hormone.
authors: Department of Neurosurgery, Teikyo University Ichihara Hospital, 3426-3 Anesaki, Ichihara City, Chiba, 299-0111, Japan. akirakun@med.teikyo-u.ac.jp
authors: Akira
authors: A
authors: Matsuno
authors: Akiko
authors: A
authors: Mizutani
authors: Susumu
authors: S
authors: Takekoshi
authors: Johbu
authors: J
authors: Itoh
authors: Hiroko
authors: H
authors: Okinaga
authors: Yuko
authors: Y
authors: Nishina
authors: Koji
authors: K
authors: Takano
authors: Tadashi
authors: T
authors: Nagashima
authors: R Yoshiyuki
authors: RY
authors: Osamura
authors: Akira
authors: A
authors: Teramoto
mesh_terms: D000818:Animals; D016375:Antisense Elements (Genetics); D001426:Bacterial Proteins; D001693:Biological Transport, Active; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D042541:Intracellular Space / Q000378:metabolism*; D008164:Luminescent Proteins; D018613:Microscopy, Confocal; D008854:Microscopy, Electron; D010907:Pituitary Hormones / Q000096:biosynthesis / Q000378:metabolism*; D045663:Quantum Dots; D012333:RNA, Messenger / Q000096:biosynthesis*; D051381:Rats; D013347:Subcellular Fractions / Q000378:metabolism / Q000648:ultrastructure
chemical_list: D016375:Antisense Elements (Genetics); D001426:Bacterial Proteins; D008164:Luminescent Proteins; D010907:Pituitary Hormones; D012333:RNA, Messenger; C054562:yellow fluorescent protein, Bacteria; D002118:Calcium