NBRP Medaka

Medaka Strain
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	Lectures of a conference

osterix-DsRed/TRAP-GFP

Strain information
Strain ID	TG1359
Strain Name	osterix-DsRed/TRAP-GFP
Strain Type	Transgenics
Characteristics	osterix-DsRed/TRAP-GFP transgenic line marks osteoblast (DsRed) and osteoclast (GFP).
Journal	4 Hits. PMID: 31315073 [ RRC reference ] Azetsu Y, Chatani M, Dodo Y, Karakawa A, Sakai N, Negishi-Koga T, Takami M. Treatment with synthetic glucocorticoid impairs bone metabolism, as revealed by in vivo imaging of osteoblasts and osteoclasts in medaka fish. Biomed Pharmacother 2019/10/01.118.109101. PMID: 28004797 [ RRC reference ] Chatani M, Morimoto H, Takeyama K, Mantoku A, Tanigawa N, Kubota K, Suzuki H, Uchida S, Tanigaki F, Shirakawa M, Gusev O, Sychev V, Takano Y, Itoh T, Kudo A. Acute transcriptional up-regulation specific to osteoblasts/osteoclasts in medaka fish immediately after exposure to microgravity. Sci Rep 2016/12/22.6.39545. PMID: 26658319 [ RRC reference ] Mantoku A, Chatani M, Aono K, Inohaya K, Kudo A. Osteoblast and osteoclast behaviors in the turnover of attachment bones during medaka tooth replacement. Dev Biol 2016/01/15.409(2).370-81. PMID: 26387549 [ RRC reference ] Chatani M, Mantoku A, Takeyama K, Abduweli D, Sugamori Y, Aoki K, Ohya K, Suzuki H, Uchida S, Sakimura T, Kono Y, Tanigaki F, Shirakawa M, Takano Y, Kudo A. Microgravity promotes osteoclast activity in medaka fish reared at the international space station. Sci Rep 2015/09/21.5.14172.
Special affairs　[Address]	■ The RECIPIENT shall give proper credit to the DEVELOPER in any publications reporting results of research using any derivatives developed from the BIOLOGICAL MATERIALS (hereinafter referred to as “the DERIVATIVES”), and shall procure other institutionsto which the DERIVATIVES are provided (including when provided through the NBRP) to do the same.
Category	Transgenic strains
Organization	National Institute for Basic Biology
Frozen sperm in NIBB	○
Deposited by	Showa University
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Image	osterix-DsRed_transgene map To generate osterix promoter-DsRed medaka (osterix-DsRed medaka), we amplified the 4.1 kb upstream regulatory region of the medaka osterix gene for generating osterix-DsRed medaka by using the following primers: Osterix promoter sal1 forward primer 5′- GTCGACTGAACATGTCAGTGCCATCA-3′and Osterix promoter sal1 reverse primer 5′- GTCGACCGGGACAGTTTGGAAGAAGT-3′. The amplified fragment was cloned into the TA cloning vector. We then digested the fragment with sal1, and subcloned it into the sal1 site of the I-SceI backbone vector, in which DsRed-Express/SV40polyA (Clontech) was inserted into the HindIII and AflII sites of pBSKI2 (AMAGEN)
Image	TRAP-GFP_transgene map.png A 3-kb upstream regulatory region of the medaka TRAP gene was amplified using the primers TRAP pro F-HindIII, 5′-CCCAAGCTTGGGTCGCACTCTGGATACAACAGC-3′,andTRAPproR,5′-TTCAACTGAAGCAACACACC-3′.Theamplified fragment was cloned into the TA cloning vector. We then digested the 5′-half fragment with HindIII and BamHI, and inserted the I-SceI backbone vector, which was inserted into EGFP/SV40polyA (Clontech) at the HindIII/AflII sites in pBSKI2 vector(AMAGEN).
Image	trap EGFP osterix DsRed Lateral view of a part of a vertebral column. Position of osteoclasts and osteoblasts are shown by TRAP-GFP and osterix-DsRed expressions, respectively. na, neural arch; ct, centrum; ha, hemal arch. (Chatani et al., Journal of the Showa University society 2017)