A role of microglia in neurovascular coupling during cerebal ischemia

• Project/Area Number: 15K06760
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Nerve anatomy/Neuropathology
• Research Institution: Keio University
• Principal Investigator: Tomita Yutaka 慶應義塾大学, 医学部(信濃町), 講師(非常勤) (60276251)
• Research Collaborator: SUZUKI Norihiro ; KANNO Iwao ; UNEKAWA Miyuki ; MASAMOTO Kazuto ; YUKI Hiroya ; SOGA Naoto ; WATANABE Tatsushi ; HATAKEYAMA Nao
• Project Period (FY): 2015-10-21 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2017: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2016: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2015: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: マイクログリア / neurovascular unit / 脳梗塞 / 二光子顕微鏡 / 微小循環 / 低酸素負荷 / 血管新生

Outline of Final Research Achievements

In genetically-modified mice (CX3CR1-GFP mice）expressing the cortical microglia with green fluorescence protein (GFP), microglia and cortical microvessels were imaged through a closed-cranial window by using a Tomita-Seylaz method with two-photon laser scanning fluorescence microscopy repeatedly up to 3 weeks. There were no detectable differences in the soma area, number density, and number of processes of the GFP-positive microglia, whereas migrated and newborn microglia could be detected. Furthermore, appearance and/or migration of microglia adjacent to neovascular capillary under chronic hypoxia (8% oxygen) was exhibited.
A stimulation with a blue light in mice genetically expressing a light-sensitive cation channel, channelrhodopsin-2 in cortical astrocytes or neurons elicited intensity-dependent increase of cerebral blood flow. Pharmacological study exhibited independent mechanism of CBF control.

Research Products

• [Journal Article] Enhanced susceptibility to cortical spreading depression in two types of Na+, K+-ATPase α2 subunit-deficient mice as a model of familial hemiplegic migraine 2. (2018)
  • Author(s): Unekawa, M. Ikeda, K., Tomita, Y., Kawakami, K. and Suzuki, N.
  • Journal Title: Cephalalgia Volume: 38 Issue: 9 Pages: 1515-1524
  • DOI: 10.1177/0333102417738249
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Tomita-Seylaz法による慢性頭窓作製後のマウス大脳皮質ミクログリアの長期反復in vivoリアルタイム解析 (2017)
  • Author(s): 結城浩弥, 正本和人, 畝川美悠紀, 冨田裕, 菅野巖, 鈴木則宏.
  • Journal Title: 脳循環代謝 Volume: 28 Pages: 249-256
  • NAID: 130006008228
  • Peer Reviewed
• [Journal Article] Dynamic diameter response of intraparenchymal penetrating arteries during cortical spreading depression and elimination of vasoreactivity to hypercapnia in anesthetized mice. (2017)
  • Author(s): Unekawa M, Tomita Y, Masamoto K, Toriumi H, Osada T, Kanno I, Suzuki N
  • Journal Title: J Cereb Blood Flow Metab Volume: 37 Issue: 2 Pages: 657-670
  • DOI: 10.1177/0271678x16636396
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Establishment and evaluation of a new highly metastatic tumor cell line 5a-D-Luc-ZsGreen expressing both luciferase and green fluorescent protein (2016)
  • Author(s): Sudo H, Tsuji A B, Sugyo A, Takuwa H, Masamoto K, Tomita Y, Suzuki N, Imamura T, Koizumi M, Saga T
  • Journal Title: Int. J. Oncol. Volume: 48 Issue: 2 Pages: 525-532
  • DOI: 10.3892/ijo.2015.3300
  • Peer Reviewed / Open Access
• [Journal Article] Long-term effects of cerebral hypoperfusion on neural density and function using misery perfusion animal model. (2016)
  • Author(s): Nishino A, Tajima Y, Takuwa H, Masamoto K, Taniguchi J, Wakizaka H, Kokuryo D, Urushihata T, Aoki I, Kanno I, Tomita Y, Suzuki N, Ikoma Y, Ito H.
  • Journal Title: Scientific reports Volume: 27 Issue: 1 Pages: 25072-25072
  • DOI: 10.1038/srep25072
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Dynamic flow velocity mapping from fluorescent dye transit times in the brain surface microcirculation of anesthetized rats and mice. (2016)
  • Author(s): Hoshikawa R, Kawaguchi H, Takuwa H, Ikoma Y, Tomita Y, Unekawa M, Suzuki N, Kanno I, Masamoto K.
  • Journal Title: Microcirculation Volume: 26 Issue: 6 Pages: 416-425
  • DOI: 10.1111/micc.12285
  • Peer Reviewed
• [Journal Article] アストロサイトによる脳血流調節 (2016)
  • Author(s): 正本和人, 畝川美悠紀, 冨田裕, 菅野巌, 鈴木則宏
  • Journal Title: 分子精神医学 Volume: 16 Pages: 17-24
• [Journal Article] Unveiling astrocytic control of cerebral blood flow with optogenetics (2015)
  • Author(s): Masamoto K, Unekawa M, Watanabe T, Toriumi H, Takuwa H, Kawaguchi H, Kanno I, Matsui K, Tanaka K F, Tomita Y, Suzuki N
  • Journal Title: Scientific reports Volume: 5 Issue: 1 Pages: 11455-11455
  • DOI: 10.1038/srep11455
  • Peer Reviewed / Open Access
• [Journal Article] Hyperperfusion counteracted by transient rapid vasoconstriction followed by long-lasting oligemia induced by cortical spreading depression in anesthetized mice. (2015)
  • Author(s): Unekawa M, Tomita Y, Toriumi H, Osada T, Masamoto K, Kawaguchi H, Itoh Y, Kanno I, Suzuki N.
  • Journal Title: J Cereb Blood Flow Metab Volume: 35 Issue: 4 Pages: 689-698
  • DOI: 10.1038/jcbfm.2014.250
  • Peer Reviewed / Open Access
• [Presentation] 大脳皮質拡延性抑制／脱分極が結ぶ片頭痛と脳卒中 (2018)
  • Author(s): 畝川美悠紀, 冨田裕, 鈴木則宏
  • Organizer: 第43回日本脳卒中学会学術集会
  • Invited
• [Presentation] Effect of spreading depolarization on cerebral blood flow and development of infarction under experimental ischemia in anesthetized mice. (2017)
  • Author(s): Unekawa M, Tomita Y, Toriumi H, Osada T, Masamoto K, Kanno I, Suzuki N
  • Organizer: 28th International Symposium on Cerebral Blood Flow, Metabolism & Function
  • Place of Presentation: ベルリン（ドイツ）
  • Year and Date: 2017-04-01
  • Int'l Joint Research
• [Presentation] Repeated longitudinal in vivo imaging of cortical microglia under chronic hypoxia in the mice using twophoton microscopy with a closed cranial window technique. (2017)
  • Author(s): Kanno I, Masamoto K, Yuki H, Sugashi T, Unekawa M, Tomita Y, Suzuki N
  • Organizer: 28th International Symposium on Cerebral Blood Flow, Metabolism & Function
  • Place of Presentation: ベルリン（ドイツ）
  • Year and Date: 2017-04-01
  • Int'l Joint Research
• [Presentation] Spatiotemporal changes in cerebral blood flow induced by transcranial photostimulation to cortical neurons or astrocytes in the transgenic mice expressing channelrhodopsin-2. (2017)
  • Author(s): Hatakeyama N, Masamoto K, Unekawa M, Takuwa H, Kanno I, Matsui K, Tanaka KF, Tomita Y, Suzuki N
  • Organizer: 28th International Symposium on Cerebral Blood Flow, Metabolism & Function
  • Place of Presentation: ベルリン（ドイツ）
  • Year and Date: 2017-04-01
  • Int'l Joint Research
• [Presentation] Longitudinal two-photon imaging of microvascular remodeling and microglial response to chronic hypoxia in in vivo mouse cortex (2017)
  • Author(s): Yuki T, Masamoto K, Unekawa M, Sugashi T, Tomita Y, Kanno I, Suzuki N
  • Organizer: 第42回日本微小循環学会総会
  • Place of Presentation: 富山国際会議場(富山県富山市)
  • Year and Date: 2017-03-25
• [Presentation] 慢性低灌流モデルマウスにおける安静時及び賦活時の脳神経機能評価 (2017)
  • Author(s): 漆畑拓弥, 田桑弘之, 高堂裕平, 正本和人, 菅野巖, 冨田裕, 鈴木則宏, 生駒洋子, 小畠隆行, 伊藤浩
  • Organizer: 第42回日本脳卒中学会学術集会
  • Place of Presentation: 大阪国際会議場(大阪府大阪市)
  • Year and Date: 2017-03-16
• [Presentation] 中大脳動脈閉塞時に発生する拡延性脱分極が脳血流と梗塞形成に及ぼす影響 (2017)
  • Author(s): 畝川美悠紀, 冨田裕, 鳥海春樹, 長田高志, 正本和人, 菅野巖, 鈴木則宏
  • Organizer: 第42回日本脳卒中学会学術集会
  • Place of Presentation: 大阪国際会議場(大阪府大阪市)
  • Year and Date: 2017-03-16
• [Presentation] 神経賦活とグリア賦活による脳血流反応の比較：オプトジェネティクスによる検討 (2017)
  • Author(s): 畠山菜緒, 正本和人, 畝川美悠紀, 田桑弘之, 菅野巖, 松井広, 田中謙二, 冨田裕, 鈴木則宏
  • Organizer: 第42回日本脳卒中学会学術集会
  • Place of Presentation: 大阪国際会議場(大阪府大阪市)
  • Year and Date: 2017-03-16
• [Presentation] 脳虚血後のグリア細胞の形状変化におけるin vivo長期解析 (2017)
  • Author(s): 渡部真子, 須貸拓馬, 蜂谷亮太, 安部貴人, 畝川美悠紀, 鳥海春樹, 冨田裕, 鈴木則宏, 田桑弘之, 菅野巖, 正本和人
  • Organizer: 日本機械学会 第30 回バイオエンジニアリング講演会
• [Presentation] 低酸素誘発性脳微小血管リモデリングにおけるミクログリアの関与 (2017)
  • Author(s): 曽我直人, 結城浩弥, 須貸拓馬, 畝川美悠紀, 冨田裕, 菅野巖, 鈴木則宏, 正本和人
  • Organizer: 酸素ダイナミクス研究会
• [Presentation] Na+,K+-ATPase a2サブユニット遺伝子欠損マウスにおける皮質拡延性抑制に対する感受性と脳内c-Fos発現分布 (2017)
  • Author(s): 畝川美悠紀, 池田啓子, 冨田裕, 川上潔, 鈴木則宏
  • Organizer: 第60回日本脳循環代謝学会学術集会
• [Presentation] Widespread c-Fos expression after cortical spreading depression; possible differences between two types of animal models of familial hemiplegic migraine 2. (2017)
  • Author(s): Unekawa M., Ikeda K., Tomita Y., Kawakami K., Suzuki N.
  • Organizer: The XXIII World Congress of Neurology
  • Int'l Joint Research
• [Presentation] Enhanced susceptibility to cortical spreading depression and different degree in two-types of Na+,K+-ATPase a2 subunit-deficient mice as a model of familial hemiplegic migraine 2. (2017)
  • Author(s): Unekawa M., Ikeda K., Tomita Y., Kawakami K., Suzuki N.
  • Organizer: 18th Congress of the International Headache Society
  • Int'l Joint Research
• [Presentation] Hemodynamic response to optogenetic activation of ChR2-expressing neurons or astrocytes in the mouse cortex. (2017)
  • Author(s): Hatakeyama N., Masamoto K., Unekawa M., Takuwa H., Kanno I., Matsui K., Tanaka K. F., Tomita Y., Suzuki N.
  • Organizer: 第9回光操作研究会
• [Presentation] Cerebral blood flow responses to photostimulation in the transgenic miceexpressing channelrhodopsin-2 in the cortical neurons or astrocytes. (2017)
  • Author(s): Hatakeyama N., Masamoto K., Unekawa M., Takuwa H., Kanno I., Matsui K., Tanaka K., Tomita Y., Suzuki N.
  • Organizer: The XXIII World Congress of Neurology
  • Int'l Joint Research
• [Presentation] Tomita-Seylaz法による慢性頭窓作成後のマウス大脳皮質ミクログリアのin vivo長期反復解析 (2016)
  • Author(s): 結城浩弥, 正本和人, 畝川美悠紀, 冨田裕, 菅野巖, 鈴木則宏
  • Organizer: 第59回日本脳循環代謝学会学術集会
  • Place of Presentation: あわぎんホール(徳島県徳島市)
  • Year and Date: 2016-11-11
• [Presentation] 一時的脳虚血時に発生する拡延性脱分極に伴う血流変化と梗塞形成に対する影響 (2016)
  • Author(s): 畝川美悠紀, 冨田裕, 鳥海春樹, 長田高志, 正本和人, 菅野巖, 鈴木則宏
  • Organizer: 第59回日本脳循環代謝学会学術集会
  • Place of Presentation: あわぎんホール(徳島県徳島市)
  • Year and Date: 2016-11-11
• [Presentation] オプトジェネティクスを用いたアストロサイト・ニューロン賦活による脳血流空間伝播の比較 (2016)
  • Author(s): 畠山菜緒, 正本和人, 畝川美悠紀, 田桑弘之, 菅野巖, 松井広, 田中謙二, 冨田裕, 鈴木則宏
  • Organizer: 第59回日本脳循環代謝学会学術集会
  • Place of Presentation: あわぎんホール(徳島県徳島市)
  • Year and Date: 2016-11-11
• [Presentation] 貧困灌流モデルマウスにおける神経細胞分布密度および脳血管・神経機能評価 (2016)
  • Author(s): 田桑弘之, 漆畑拓弥, 田島洋佑, 西野明日香, 正本和人, 菅野巖, 冨田裕, 鈴木則宏, 生駒洋子, 伊藤浩
  • Organizer: 第59回日本脳循環代謝学会学術集会
  • Place of Presentation: あわぎんホール(徳島県徳島市)
  • Year and Date: 2016-11-11
• [Presentation] Acceleration of red blood cells in intraparenchymal capillaries and dilation of arterial diameter in response to hypercapnia were diminished after passage of cortical spreading depression in anesthetized mice. (2016)
  • Author(s): Tomita Y, Unekawa M, Toriumi H, Osada T, Masamoto K, Kawaguchi H, Itoh Y, Kanno I, Suzuki N.
  • Organizer: 第41回日本微小循環学会総会
  • Place of Presentation: コクヨホール(東京都港区)
  • Year and Date: 2016-09-23
• [Presentation] Dynamic flow velocity mapping based on transient time measurements of fluorescent dye (2016)
  • Author(s): Hoshikawa R, Kawaguchi H, Takuwa H, Ikoma Y, Tomita Y, Unekawa M, Suzuki N, Kanno I, Masamoto K.
  • Organizer: 第41回日本微小循環学会総会
  • Place of Presentation: コクヨホール(東京都港区)
  • Year and Date: 2016-09-23
• [Presentation] Cerebral microvascular restructing and microglial adaptation to chronic hypoxia in the animal models. (2016)
  • Author(s): Masamoto K, Kanno I, Tomita Y, Suzuki N.
  • Organizer: 第41回日本微小循環学会総会
  • Place of Presentation: コクヨホール(東京都港区)
  • Year and Date: 2016-09-23
  • Invited
• [Presentation] In vivo imaging and quantification of glucose transfer using 2-NBDG in the mouse cortex with two-photon microscopy. (2016)
  • Author(s): Takeda H, Suzuki H, Murata R, Takuwa H, Kanno I, Tomita Y, Suzuki N, Yamada K, Masamoto K.
  • Organizer: 第41回日本微小循環学会総会
  • Place of Presentation: コクヨホール(東京都港区)
  • Year and Date: 2016-09-23
• [Presentation] Heterogeneous response of microcirculation to spreading depolarization evoked during middle cerebral artery occlusion in anesthetized mice. (2016)
  • Author(s): Unekawa M, Tomita Y, Toriumi H, Osada T, Masamoto K, Kanno I, Suzuki N.
  • Organizer: 第41回日本微小循環学会総会
  • Place of Presentation: コクヨホール(東京都港区)
  • Year and Date: 2016-09-23
• [Presentation] マウス大脳における神経細胞の空間分布と機能活動に関する定量解析 (2016)
  • Author(s): 武田寛史, 田桑弘之, 季斌, 佐原成彦, 冨田裕, 鈴木則宏, 菅野巖, 須原哲也, 樋口真人, 正本和人
  • Organizer: 第25回日本バイオイメージング学会学術集会
  • Place of Presentation: 名古屋市立大学薬学部 宮田専治記念ホール(愛知県名古屋市)
  • Year and Date: 2016-09-05
• [Presentation] Response of cerebral blood flow to optogenetic photostimulation of cortical neurons and astrocytes (2016)
  • Author(s): 冨田裕, 渡部竜志, 正本和人, 畝川美悠紀, 鳥海春樹, 田桑弘之, 菅野巌, 松井広, 田中謙二, 鈴木則宏
  • Organizer: 第57回日本神経学会学術大会
  • Place of Presentation: 神戸コンベンションセンター(兵庫県神戸市)
  • Year and Date: 2016-05-18
• [Presentation] Response of cerebral blood flow to optogenetic photostimulation of cortical neurons and astrocytes (2016)
  • Author(s): 冨田裕, 渡部竜志, 正本和人, 畝川美悠紀, 鳥海春樹, 田桑弘之, 菅野巌, 松井広, 田中謙二, 鈴木則宏
  • Organizer: 第57回日本神経学会学術大会(神戸)
  • Place of Presentation: 神戸コンベンションセンター(兵庫県神戸市)
  • Year and Date: 2016-05-18
• [Presentation] 生体二光子顕微鏡法を用いたグリア脳血管の長期相互作用の解析 (2016)
  • Author(s): 新タ雅啓, 中原智美, 星川椋, 結城浩弥, 田桑弘之, 冨田裕, 鈴木則宏, 菅野巖, 正本和人
  • Organizer: 第25回日本神経科学大会
  • Place of Presentation: パシフィコ横浜(神奈川県横浜市)
• [Presentation] オプトジェネティクスを用いた脳賦活による脳血流反応の空間伝播解析 (2015)
  • Author(s): 渡部竜志, 正本和人, 畝川美悠紀, 冨田裕, 田桑弘之, 菅野巖, 松井広, 田中謙二, 鈴木則宏
  • Organizer: 第4回ニューロフォトニクス研究会
  • Place of Presentation: ホテルグランドヒル市ヶ谷(東京都新宿区)
  • Year and Date: 2015-12-03
• [Presentation] 二光子顕微鏡を用いた生体マウス脳内ミクログリアの三次元長期イメージングと定量解析 (2015)
  • Author(s): 結城浩弥, 正本和人, 畝川美悠紀, 冨田裕, 鳥海春樹, 海老根妙子, 菅野巖, 鈴木則宏
  • Organizer: 第4回ニューロフォトニクス研究会
  • Place of Presentation: ホテルグランドヒル市ヶ谷(東京都新宿区)
  • Year and Date: 2015-12-03
• [Presentation] 家族性片麻痺性片頭痛２型モデルマウスにおける皮質拡延性抑制感受性および反応性 (2015)
  • Author(s): 畝川美悠紀, 池田啓子, 冨田裕, 鳥海春樹, 海老根妙子, 川上潔, 鈴木則宏
  • Organizer: 第43回日本頭痛学会総会
  • Place of Presentation: 京王プラザホテル(東京都新宿区)
  • Year and Date: 2015-11-13
• [Presentation] 大脳皮質性拡延性抑制が実質内穿通動脈口径と局所脳血流および二酸化炭素負荷に対する反応性に及ぼす影響 (2015)
  • Author(s): 畝川美悠紀, 冨田裕, 鳥海春樹, 長田高志, 正本和人, 菅野巖, 鈴木則宏
  • Organizer: 第27回日本脳循環代謝学会総会
  • Place of Presentation: 富山国際会議場(富山県富山市)
  • Year and Date: 2015-10-30