Analysis of axon collaterals of cortical projection neurons by Tet double infection (TEDI) method

• Project/Area Number: 25430018
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Neurophysiology / General neuroscience
• Research Institution: Institute of Physical and Chemical Research (2015); National Institute for Basic Biology (2013-2014)
• Principal Investigator: Akiya Watakabe 国立研究開発法人理化学研究所, 脳科学総合研究センター, 研究員 (40290910)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000); Fiscal Year 2015: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2014: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000); Fiscal Year 2013: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: 大脳皮質 / 逆行性 / 神経解剖学 / マーモセット / ウイルスベクター / 投射ニューロン / 視床 / 層構造 / AAV / 逆行性ウイルス / 神経回路 / 遺伝子導入 / 領野 / レンチウイルス / TET-OFF / 皮質間投射 / 視床投射 / 錐体細胞

Outline of Final Research Achievements

In this project, I endeavored to establish and further polish up my new viral technology termed TEDI (TET double Infection method), in which retrograde and anterograde viral vectors were combined to selectively transduce a particular type of projection neurons. Using this technique, I was able to selectively visualize and analyze the morphologies of the axon collaterals of the corticothalamic and corticocortical neurons in mice. I also examined the infection conditions of various serotypes of AAV in marmosets, which result is useful in planning an efficient TEDI in non-human primates.

Research Products

• [Journal Article] In situ hybridization analyses of claustrum-enriched genes in marmosets. (2016)
  • Author(s): Watakabe A.
  • Journal Title: J Comp Neurol Volume: x Issue: 6 Pages: 1442-1458
  • DOI: 10.1002/cne.24021
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Long-term two-photon calcium imaging of neuronal populations with subcellular resolution in adult non-human primates. (2015)
  • Author(s): Sadakane O., Masamizu Y., Watakabe A., Terada S., Ohtsuka M., Takaji M., Mizukami H., Ozawa K., Kawasaki H., Matsuzaki M., Yamamori T.
  • Journal Title: Cell Reports Volume: 13 Issue: 9 Pages: 1989-1999
  • DOI: 10.1016/j.celrep.2015.10.050
  • NAID: 120005829368
  • Peer Reviewed / Open Access
• [Journal Article] in vivo two-photon imaging of dendritic spines in marmoset neocortex. (2015)
  • Author(s): Osamu Sadakane, Akiya Watakabe, Masanari Ohtsuka, Masafumi Takaji, Tetsuya Sasaki, Masatoshi Kasai, Tadashi Isa, Go Kato, Junichi Nabekura, Hiroaki Mizukami, Keiya Ozawa, Hiroshi Kawasaki, and Tetsuo Yamamori
  • Journal Title: eNeuro Volume: 2(4) Issue: 4 Pages: 1-10
  • DOI: 10.1523/eneuro.0019-15.2015
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Comparative analyses of adeno-associated viral vector serotypes 1, 2, 5, 8 and 9 in marmoset, mouse and macaque cerebral cortex. (2015)
  • Author(s): Watakabe A, Ohtsuka M, Kinoshita M, Takaji M, Isa K, Mizukami H, Ozawa K, Isa T, and Yamamori T.
  • Journal Title: Neurosci. Res. Volume: 93 Pages: 144-157
  • DOI: 10.1016/j.neures.2014.09.002
  • Peer Reviewed / Open Access
• [Journal Article] Simultaneous visualization of extrinsic and intrinsic axon collaterals in Golgi-like detail for mouse corticothalamic and corticocortical cells: a double viral infection method (2014)
  • Author(s): Watakabe, A., Takaji, M., Kato, S., Kobayashi, K., Mizukami, H., Ozawa, K., Ohsawa, S., Matsui, R., Watanabe, D., and Yamamori, T.
  • Journal Title: Frontiers in neural circuits Volume: 8 Pages: 110-110
  • DOI: 10.3389/fncir.2014.00110
  • Peer Reviewed / Open Access
• [Journal Article] Characterization of claustral neurons by comparative gene expression profiling and dye-injection analyses. Front Syst Neurosci, (2014)
  • Author(s): Watakabe A, Ohsawa S, Ichinohe N, Rockland KS, Yamamori T.
  • Journal Title: Front Syst Neurosci, Volume: 8 (98)00098-, Pages: 98-98
  • DOI: 10.3389/fnsys.2014.00098
  • Peer Reviewed / Open Access
• [Presentation] Two-photon calcium imaging using genetically-encoded calcium indicator in primate neocortex (2015)
  • Author(s): *O. SADAKANE1,2, Y. MASAMIZU3, A. WATAKABE1,2, S.-I. TERADA3,4, M. OHTSUKA1,2, M. TAKAJI1,2, H. MIZUKAMI5, K. OZAWA5,6, H. KAWASAKI7, M. MATSUZAKI3, T. YAMAMORI1,2;
  • Organizer: 北米神経学会2015
  • Place of Presentation: シカゴ・アメリカ
  • Year and Date: 2015-10-18
  • Int'l Joint Research
• [Presentation] マーモセット大脳皮質におけるGCaMPを用いた2光子カルシウムイメージング (2015)
  • Author(s): 定金 理、正水 芳人 、渡我部 昭哉、寺田 晋一郎 、大塚 正成、高司 雅史、水上 浩明、小澤 敬也、河崎 洋志、松崎 政紀、山森 哲雄
  • Organizer: 第３８回日本神経科学大会
  • Place of Presentation: 神戸国際会議場・神戸市・兵庫県
  • Year and Date: 2015-07-29
• [Presentation] Corticothalamic and corticocortical cells in mouse M1 exhibit layer- and area-specific elaboration of axon collaterals (2014)
  • Author(s): Watakabe A, Takaji M, Kato S, Kobayashi K, Mizukami H, Ozawa K, Ohsawa S, Matsui R, Watanabe D, Yamamori T.
  • Organizer: 北米神経学会
  • Place of Presentation: 米国ワシントンDC
  • Year and Date: 2014-11-15 – 2014-11-19
• [Presentation] TET2重感染法(TEDI)を使った大脳皮質投射ニューロンサブタイプの同時標識
  • Author(s): 渡我部 昭哉、加藤 成樹、小林 和人、水上 浩明、小澤 敬也、山森 哲雄
  • Organizer: 日本神経科学学会( Neuro2013）
  • Place of Presentation: 京都国際会館