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齊藤 秀俊(さいとう ひでとし) データ更新日:2019.08.01

准教授 /  薬学研究院 臨床薬学部門 ライフイノベーション分野


主な研究テーマ
慢性疼痛モデル病態へのミクログリアの関与
キーワード:microglia, choronic pain
2015.06~2019.01.
神経障害性疼痛発症における脊髄グリア細胞の役割の解明
キーワード:神経障害性疼痛、グリア、ATP受容体、多光子励起イメージング
2006.04.
研究業績
主要原著論文
1. Hidetoshi Saitoh, Junya Masuda, Ryu Kawada, Chinami Kojima, Sosuke Yoneda, Takahiro Masuda, Kazuhide Inoue, Tsuda Makoto, Transcription factor MafB contributes to the activation of spinal microglia underlying neuropathic pain development, GLIA, 10.1002/glia.23570, 67, 4, 729-740, 2019.04, Microglia, which are pathological effectors and amplifiers in the central nervous system, undergo various forms of activation. A well-studied microglial-induced pathological paradigm, spinal microglial activation following peripheral nerve injury (PNI), is a key event for the development of neuropathic pain but the transcription factors contributing to microglial activation are less understood. Herein, we demonstrate that MafB, a dominant transcriptional regulator of mature microglia, is involved in the pathology of a mouse model of neuropathic pain. PNI caused a rapid and marked increase of MafB expression selectively in spinal microglia but not in neurons. We also found that the microRNA mir-152 in the spinal cord which targets MafB expression decreased after PNI, and intrathecal administration of mir-152 mimic suppressed the development of neuropathic pain. Reduced MafB expression using heterozygous Mafb deficient mice and by intrathecal administration of siRNA alleviated the development of PNI-induced mechanical hypersensitivity. Furthermore, we found that intrathecal transfer of Mafb deficient microglia did not induce mechanical hypersensitivity and that conditional Mafb knockout mice did not develop neuropathic pain after PNI. We propose that MafB is a key mediator of the PNI-induced phenotypic alteration of spinal microglia and neuropathic pain development..
2. Katsuyuki Matsushita, Hidetoshi Saitoh, Chinami Kojima, Masuda Takahiro, TSUDA MAKOTO, Kazuhide Inoue, Sumio Hoka, Chemokine (C-C motif) Receptor 5 Is an Important Pathological Regulator in the Development and Maintenance of Neuropathic Pain., Anesthesiology, 120, 6, 1491-503, 2014.06, BACKGROUND:
The chemokine family has been revealed to be involved in the pathogenesis of neuropathic pain. In this study, the authors investigated the role of chemokine (C-C motif) ligand 3 and its receptors chemokine (C-C motif) receptor 1 and chemokine (C-C motif) receptor (CCR) 5 in neuropathic pain.
METHODS:
A spinal nerve injury model was established in adult male Wistar rats. The von Frey test and hot plate test were performed to evaluate neuropathic pain behavior, and real-time quantitative reverse transcription polymerase chain reaction, in situ hybridization, and immunohistochemistry were performed to understand the molecular mechanisms.
RESULTS:
The expression levels of chemokine (C-C motif) ligand 3 and CCR5 messenger RNA in the spinal cord were up-regulated after nerve injury, which was possibly due to CD11b-positive microglia. Single intrathecal administration of recombinant chemokine (C-C motif) ligand 3 produced biphasic tactile allodynia; each phase of pain behavior was induced by different receptors. Intrathecal injection of CCR5 antagonist suppressed the development of tactile allodynia (12.81 ± 1.33 g vs. 3.52 ± 0.41 g [mean ± SEM, drug vs. control in paw-withdrawal threshold]; P < 0.05, n = 6 each) and could reverse established tactile allodynia (10.87 ± 0.91 g vs. 3.43 ± 0.28 g; P < 0.05, n = 8 and 7). Furthermore, Oral administration of CCR5 antagonist could reverse established tactile allodynia (8.20 ± 1.27 g vs. 3.18 ± 0.46 g; P < 0.05, n = 4 each).
CONCLUSIONS:
Pharmacological blockade of CCR5 was effective in the treatment of the development and maintenance phases of neuropathic pain. Thus, CCR5 antagonists may be potential new drugs for the treatment of neuropathic pain..
3. Uesugi A, Kataoka A, Tozaki-Saitoh H, Koga Y, Tsuda M, Robaye B, Boeynaems JM, Inoue K., Involvement of protein kinase D in uridine diphosphate-induced microglial macropinocytosis and phagocytosis., Glia, 60, 7, 1094-105, 2012.07, The clearance of tissue debris by microglia is a crucial component of maintaining brain homeostasis. Microglia continuously survey the brain parenchyma and utilize extracellular nucleotides to trigger the initiation of their dynamic responses. Extracellular uridine diphosphate (UDP), which leaks or is released from damaged neurons, has been reported to stimulate the phagocytotic activity of microglia through P2Y(6) receptor activation. However, the intracellular mechanisms underlying microglial P2Y(6) receptor signals have not been identified. In this study, we demonstrated that UDP stimulation induced immediate and long-lasting dynamic movements in the cell membrane. After 60 min of UDP stimulation, there was an upregulation in the number of large vacuoles formed in the cell that incorporate extracellular fluorescent-labeled dextran, which indicates microglial macropinocytosis. In addition, UDP-induced vacuole formation and continuous membrane motility were suppressed by the protein kinase D (PKD) inhibitors, Gö6976 and CID755673, unlike Gö6983, which is far less sensitive to PKD. The inhibition of PKD also reduced UDP-induced incorporation of fluorescent-labeled dextran and soluble β-amyloid and phagocytosis of microspheres. UDP induced rapid phosphorylation and membrane translocation of PKD, which was abrogated by the inhibition of protein kinase C (PKC) with Gö6983. However, Gö6983 failed to suppress UDP-induced incorporation of microspheres. Finally, we found that inhibition of PKD by CID755673 significantly suppressed UDP-induced engulfment of IgG-opsonized microspheres. These data suggest that a PKC-independent function of PKD regulates UDP-induced membrane movement and contributes to the increased uptake of extracellular fluid and microspheres in microglia..
4. Toyomitsu E, Tsuda M, Yamashita T, Tozaki-Saitoh H, Tanaka Y, Inoue K., CCL2 promotes P2X4 receptor trafficking to the cell surface of microglia., Purinergic Signaling, 8, 2, 301-10, 2012.06, P2X4 receptors (P2X4Rs), a subtype of the purinergic P2X family, play important roles in regulating neuronal and glial functions in the nervous system. We have previously shown that the expression of P2X4Rs is upregulated in activated microglia after peripheral nerve injury and that activation of the receptors by extracellular ATP is crucial for maintaining nerve injury-induced pain hypersensitivity. However, the regulation of P2X4R expression on the cell surface of microglia is poorly understood. Here, we identify the CC chemokine receptor CCR2 as a regulator of P2X4R trafficking to the cell surface of microglia. In a quantitative cell surface biotinylation assay, we found that applying CCL2 or CCL12, endogenous ligands for CCR2, to primary cultured microglial cells, increased the levels of P2X4R protein on the cell surface without changing total cellular expression. This effect of CCL2 was prevented by an antagonist of CCR2. Time-lapse imaging of green fluorescent protein (GFP)-tagged P2X4R in living microglial cells showed that CCL2 stimulation increased the movement of P2X4R-GFP particles. The subcellular localization of P2X4R immunofluorescence was restricted to lysosomes around the perinuclear region. Notably, CCL2 changed the distribution of lysosomes with P2X4R immunofluorescence within microglial cells and induced release of the lysosomal enzyme β-hexosaminidase, indicating lysosomal exocytosis. Moreover, CCL2-stimulated microglia enhanced Akt phosphorylation by ATP applied extracellularly, a P2X4R-mediated response. These results indicate that CCL2 promotes expression of P2X4R protein on the cell surface of microglia through exocytosis of P2X4R-containing lysosomes, which may be a possible mechanism for pain hypersensitivity after nerve injury..
5. Tsuda M, Tozaki-Saitoh H, Inoue K., Platelet-activating factor and pain., Biological and pharmaceutical bulletin, 34, 8, 1159-62, 2012.06, Platelet-activating factor (PAF) is a phospholipid mediator that regulates the functions of a variety of cells in the peripheral tissues and in the nervous system. Findings that injection of PAF exogenously at the skin or in the spinal cord induced pain hypersensitivity gave us much attention to its role in pain signaling. Studies using pharmacological and genetic tools to control the functions of the PAF receptor (PAFR) revealed that the PAF/PAFR system plays a role in tissue injury-induced pain, but not in the acute physiological pain evoked by thermal and mechanical stimuli. Recent investigations have focused on the roles of PAFR in pathological chronic pain such as the neuropathic pain that occurs after nerve injury for which there is currently no effective therapy. Nerve injury upregulated PAFRs in dorsal root ganglion (DRG) neurons. Studies using PAFR antagonists and PAFR-deficient mice indicated a crucial role of PAFR in production of tumor necrosis factor α (TNFα) and interleukin-1β (IL-1β) in the DRG and in developing and maintaining neuropathic pain. Thus, blocking PAFRs may be a viable therapeutic strategy for treating neuropathic pain..
6. Kataoka A, Koga Y, Uesugi A, Tozaki-Saitoh H, Tsuda M, Inoue K., Involvement of vasodilator-stimulated phosphoprotein in UDP-induced microglial actin aggregation via PKC- and Rho-dependent pathways., Purinergic Signaling, 7, 4, 403-11, 2011.12, Microglia are major immunocompetent cells in the central nervous system and retain highly dynamic motility. The processes which allow these cells to move, such as chemotaxis and phagocytosis, are considered part of their functions and are closely related to purinergic signaling. Previously, we reported that the activation of the P2Y(6) receptor by UDP stimulation in microglia evoked dynamic cell motility which enhanced their phagocytic capacity, as reported by Koizumi et al. (Nature 446(7139):1091-1095, 2007). These responses require actin cytoskeletal rearrangement, which is seen after UDP stimulation. However, the intracellular signaling pathway has not been defined. In this study, we found that UDP in rat primary microglia rapidly induced the transient phosphorylation at Ser157 of vasodilator-stimulated phosphoprotein (VASP). VASP, one of actin binding protein, accumulated at the plasma membrane where filamentous (F)-actin aggregated in a time-dependent manner. The phosphorylation of VASP was suppressed by inhibition of PKC. UDP-induced local actin aggregations were also abrogated by PKC inhibitors. The Rho inhibitor CT04 and the expression of p115-RGS, which suppresses G(12/13) signaling, attenuated UDP-induced phosphorylation of VASP and actin aggregation. These results indicate that PKC- and Rho-dependent phosphorylation of VASP is involved in UDP-induced actin aggregation of microglia..
7. Tozaki-Saitoh H, Tsuda M, Inoue K., Role of purinergic receptors in CNS function and neuroprotection., Advances in pharmacology , 61, 495-528, 2011.10, The purinergic receptor family contains some of the most abundant receptors in living organisms. A growing body of evidence indicates that extracellular nucleotides play important roles in the regulation of neuronal and glial functions in the nervous system through purinergic receptors. Nucleotides are released from or leaked through nonexcitable cells and neurons during normal physiological and pathophysiological conditions. Ionotropic P2X and metabotropic P2Y purinergic receptors are expressed in the central nervous system (CNS), participate in the synaptic processes, and mediate intercellular communications between neuron and gila and between glia and other glia. Glial cells in the CNS are classified into astrocytes, oligodendrocytes, and microglia. Astrocytes express many types of purinergic receptors, which are integral to their activation. Astrocytes release adenosine triphosphate (ATP) as a "gliotransmitter" that allows communication with neurons, the vascular walls of capillaries, oligodendrocytes, and microglia. Oligodendrocytes are myelin-forming cells that construct insulating layers of myelin sheets around axons, and using purinergic receptor signaling for their development and for myelination. Microglia also express many types of purinergic receptors and are known to function as immunocompetent cells in the CNS. ATP and other nucleotides work as "warning molecules" especially by activating microglia in pathophysiological conditions. Studies on purinergic signaling could facilitate the development of novel therapeutic strategies for disorder of the CNS..
8. Kuboyama K, Harada H, Tozaki-Saitoh H, Tsuda M, Ushijima K, Inoue K., Astrocytic P2Y(1) receptor is involved in the regulation of cytokine/chemokine transcription and cerebral damage in a rat model of cerebral ischemia., Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism., 10.1038/jcbfm.2011.49, 31, 9, 1930-41, 2011.09, After brain ischemia, significant amounts of adenosine 5'-triphosphate are released or leaked from damaged cells, thus activating purinergic receptors in the central nervous system. A number of P2X/P2Y receptors have been implicated in ischemic conditions, but to date the P2Y(1) receptor (P2Y(1)R) has not been implicated in cerebral ischemia. In this study, we found that the astrocytic P2Y(1)R, via phosphorylated-RelA (p-RelA), has a negative effect during cerebral ischemia/reperfusion. Intracerebroventricular administration of the P2Y(1)R agonist, MRS 2365, led to an increase in cerebral infarct volume 72 hours after transient middle cerebral artery occlusion (tMCAO). Administration of the P2Y(1)R antagonist, MRS 2179, significantly decreased infarct volume and led to recovered motor coordination. The effects of MRS 2179 occurred within 24 hours of tMCAO, and also markedly reduced the expression of p-RelA and interleukin-6, tumor necrosis factor-α, monocyte chemotactic protein-1/chemokine (C-C motif) ligand 2 (CCL2), and interferon-inducible protein-10/chemokine (C-X-C motif) ligand 10 (CXCL10) mRNA. P2Y(1)R and p-RelA were colocalized in glial fibrillary acidic protein-positive astrocytes, and an increase in infarct volume after MRS 2365 treatment was inhibited by the nuclear factor (NF)-κB inhibitor ammonium pyrrolidine dithiocarbamate. These results provide evidence that the P2Y(1)R expressed in cortical astrocytes may help regulate the cytokine/chemokine response after tMCAO/reperfusion through a p-RelA-mediated NF-κB pathway.
9. Biber K, Tsuda M, Tozaki-Saitoh H, Tsukamoto K, Toyomitsu E, Masuda T, Boddeke H, Inoue K. , Neuronal CCL21 up-regulates microglia P2X4 expression and initiates neuropathic pain development, EMBO Journal, 30, 9, 1864-1873, 2011.05, 末梢神経傷害後に引き起こされる疼痛は神経障害性疼痛と呼ばれ、その病態生理学的なメカニズムは未だ解明に至っていない。有効な治療法の確立には発症メカニズムの解明が必要であるが、当報告ではまず物理的に傷害を受けた一次求心性神経からCCL21と呼ばれるタンパク質が産生され、神経の投射先である脊髄後角に運搬されていることを見出した。CCL21を持たないマウスでは疼痛発症は顕著に抑制され、CCL21を外因的に添加することによって疼痛の形成が見られた。脊髄組織内の細胞を詳細に解析したところCCL21はミクログリアと呼ばれる細胞内のP2X4受容体の発現を亢進する作用を持つことがわかり、傷害された神経から脊髄内ミクログリアの活性化までの一連のプロセスを解明した。.
10. Maeda M, Tsuda M, Tozaki-Saitoh H, Inoue K, Kiyama H. , Nerve injury-activated microglia engulf myelinated axons in a P2Y12 signaling-dependent manner in the dorsal horn, Glia, 58, 15, 1838-1846, 2010.11.
11. Tozaki-Saitoh H, Tsuda M, Inoue K. , Purinergic regulation of microglia
, 日本薬理学雑誌, 136, 2, 93-97,
, 2010.08.
12. Shiratori M, Tozaki-Saitoh H, Yoshitake M, Tsuda M, Inoue K., P2X7 receptor activation induces CXCL2 production in microglia through NFAT and PKC/MAPK pathways., Journal of Neurochemistry, 10.1111/j.1471-4159.2010.06809.x, 114, 3, 810-9, 2010.08.
13. Tsuda M, Tozaki-Saitoh H, Inoue K. , Pain and purinergic signaling.
, Brain Research Review, 63, 1-2, 222-232,
, 2010.05.
14. Tsuda M, Tozaki-Saitoh H, Inoue K., Pain and purinergic signaling., Brain Research Reviews, 10.1016/j.brainresrev.2009.11.003, 63, 1-2, 222-232 , 2010.05.
15. Tsuda M, Masuda T, Kitano J, Shimoyama H, Tozaki-Saitoh H, Inoue K., IFN-{gamma} receptor signaling mediates spinal microglia activation driving neuropathic pain., Proc Natl Acad Sci U S A., 2009.04.
16. Kataoka A, Tozaki-Saitoh H, Koga Y, Tsuda M, Inoue K., Activation of P2X receptors induces CCL3 production in microglial cells through transcription factor NFAT.
, J Neurochem., 108(1):115-25, 2009.01.
17. Tozaki-Saitoh H, Tsuda M, Miyata H, Ueda K, Kohsaka S, Inoue K. , P2Y12 Receptors in Spinal Microglia Are Required for Neuropathic Pain after Peripheral Nerve Injury., The Journal of Neuroscience, 28(19):4949-56, 2008.05.
18. Tozaki-Saitoh H, Koizumi S, Sato Y, Tsuda M, Nagao T, Inoue K., Retinoic acids increase P2X2 receptor expression through the 5'-flanking region of P2rx2 gene in rat phaeochromocytoma PC-12 cells., Molecullar Pharmacology, 70(1),319-28., 2006.07.
主要総説, 論評, 解説, 書評, 報告書等
1. 齊藤 秀俊, 津田 誠, 井上 和秀, 慢性痛とグリア細胞, Clinical Neuroscience, 2015.07.
2. 齊藤 秀俊, 津田 誠, 井上 和秀, P2Y 受容体(ATP 受容体)の生理的および薬理的作用, Clinical Neuroscience, 2015.05.
3. 齊藤 秀俊, 津田 誠, 井上 和秀, P2Y 受容体(ATP 受容体)の種類, Clinical Neuroscience, 2015.04.
4. 齊藤 秀俊, 津田 誠, 井上 和秀, P2Y受容体(ATP 受容体)関連分子をターゲットとした臨床応用, Clinical Neuroscience, 2015.06.
5. Tsuda M, Tozaki-Saitoh H, Inoue K., Purinergic system, microglia and neuropathic pain., Current opinion in Pharmacology, 2012.02, Extracellular nucleotides play pivotal roles in the regulation of neuronal and glial functions in the nervous system through P2X receptors (P2XRs) and P2Y receptors (P2YRs). A growing body of evidence shows that microglia express several subtypes of P2XRs and P2YRs, and that these receptors play a key role in pain signaling in the spinal cord under pathological conditions, such as following peripheral nerve injury (neuropathic pain). Following peripheral nerve injury, dorsal horn microglia become activated and show upregulated expression of purinergic receptors, and interference with the function or expression of these receptors strongly suppresses neuropathic pain. This article highlights recent advances that further increase our understanding of the mechanisms by which microglial purinergic receptors contribute to the pathogenesis of neuropathic pain..
6. 津田誠、齊藤秀俊、井上和秀, 神経障害性疼痛におけるATP受容体の役割, ペインクリニック, 2008.02.
主要学会発表等
1. 齊藤秀俊、八田拓哉、津田誠, Social defeat stress modulates nocifensive response in remission phase of chronic pain models., 第42回日本神経科学大会/第62回日本神経化学会大会 (NEURO2019), 2019.07.
2. 齊藤秀俊,津田誠, 心理社会的ストレスによる慢性疼痛の変調:ミクログリアの関与
Modulation of pain chronicity by psychosocial stress: an involvement of microglia
, 第115回日本精神神経学会学術総会, 2019.06.
3. 齊藤秀俊、増田潤哉、米田聡介、小嶋ちなみ、川田竜、井上和秀、津田誠, 転写因子MafBは脊髄ミクログリアの活性化と神経障害性疼痛に関与する, 第92回日本薬理学会年会, 2019.03.
4. 齊藤秀俊、増田潤哉、川田竜、小嶋ちなみ、米田聡介、井上和秀、津田誠, 脊髄後角ミクログリアの転写因子MafBは末梢神経障害後の神経障害性疼痛発症に関与する, 第35回日本薬学会九州支部大会, 2018.11.
5. 笹木泉、齊藤秀俊、津田誠, アドレナリン受容体を介したミクログリアの炎症応答調節, 第35回日本薬学会九州支部大会, 2018.11.
6. 齊藤秀俊,八田拓哉,井上和秀,津田誠, 神経障害性疼痛寛解期の社会的敗北ストレス負荷は痛覚閾値の再低下を引き起こす, 第71回日本薬理学会西南部会, 2018.11.
7. Hidetoshi Saitoh, Takuya Hatta, Kazuhide Inoue, Makoto Tsuda, Social defeat stress arouses recurrence of pain in ameliorationg state, 18th World Congress of Basic and Clinical Pharmacology (WCP2018), 2018.07.
8. 齊藤秀俊, 増田潤哉, 米田聡介, 小嶋ちなみ, 川田竜, 井上和秀, 津田誠, 神経障害性疼痛モデルにおいて脊髄ミクログリアの活性化に関与する転写因子, 第27回神経行動薬理若手研究者の集い, 2018.06.
9. 齊藤 秀俊, 井上 和秀, 津田 誠, 転写因子MafBは末梢神経障害後の脊髄後角ミクログリアの活性化を調節し神経障害性疼痛に関与する, 第39回日本疼痛学会, 2017.06.
10. 齊藤 秀俊, 増田潤哉, 小嶋ちなみ, 川田竜, 津田 誠, 井上 和秀, Transcription factor MafB mediates activation process of spinal microglia that contributes to neuropathic pain development, 第90回日本薬理学会年会, 2017.03.
11. 齊藤 秀俊, Kazushi Tachibana, 井上 和秀, 津田 誠, 転写因子IRF8によるミクログリアの発達制御と中枢神経系への影響, 第26回神経行動薬理若手研究者の集い, 2017.03.
12. 松田 烈士, 齊藤 秀俊, 津田 誠, 井上 和秀, 侵害刺激時における脊髄アストロサイトシグナリングのin vivo imaging, 第67回日本薬理学会西南部会, 2014.11.
13. 松田烈士, 増田 隆博, 西本奈央, 齊藤 秀俊, 津田 誠, 井上 和秀, 高坂新一, 田村智彦, Transcriptional regulation of microglial motility by IRF8, 第87回日本薬理学会年会, 2014.03, Microglia are the resident immune cells of the central nervous system (CNS). In the response to the tissue damage in the CNS, microglia immediately trigger directional movement toward the site of injury. This microglial responses have been known to be regulated by purinergic signaling. Recently, we identified the transcription factor interferon regulatory factor 8 (IRF8) is expressed specifically in microglia in the CNS, and play a central role in regulating microglial gene expression(e.g. Tlr2, P2ry12, and Cx3cr1). However, whether IRF8 participates in microglial cell motility is remained unclear. Here we report that IRF8 may contribute to microglial motility by controlling an expression pattern of microglial genes..
14. 松下克之, 齊藤 秀俊, 小嶋ちなみ, 津田 誠, 井上 和秀, C-C chemokine receptor type 5 is an important pathological regulator in the development and maintenance of neuropathic pain, 第87回日本薬理学会年会, 2014.03, Neuropathic pain is a reflection of aberrant functioning of pathologically altered nervous systems, which is often caused by peripheral nerve injury and represented by one hallmark called tactile allodynia (pain hypersensitivity to innocuous stimuli). The mechanism underlying development of tactile allodynia remains largely unknown. Recent studies have revealed that several molecules of chemokine family is involved in the pathogenesis of neuropathic pain. However, chemokine family and also chemokine receptor family consist of large number of molecules, and there are complicated signal crosstalk between each ligand and receptor set.
Here we found that single intrathecal administration of recombinant CC chemokine ligand 3 (CCL3) produce two phase tactile allodynia-like behavior. The first phase clearly suppressed by a C-C chemokine receptor type 1 (CCR1) antagonist pretreatment but second phase was not affected. The second phase was dose-dependently suppressed by a C-C chemokine receptor type 5 (CCR5) antagonist that treated simultaneously with CCL3. In the model of neuropathic pain that caused by surgically injured peripheral nerve, the level of CCL3 and CCR5 mRNA but not CCR1 mRNA was significantly increased in ipsilateral spinal cord after nerve injury. This increase was observed from day1 to day14 after nerve injury. Intrathecal injection of the CCR5 antagonist and CCL3 neutralizing antibody successfully suppressed the development of neuropathic pain. Furthermore, we found the CCR5 antagonist can reverse the established neuropathic pain.
These results suggest that one of the chemokine signaling triggered by CCL3 has crucial role in the mechanism of neuropathic pain. Thus, CCR5 might be a effective therapeutic target for pain induced after nerve injury..
15. 小嶋ちなみ, 齊藤 秀俊, 津田 誠, 井上 和秀, CCL21による脊髄ミクログリアのMafB発現誘導, 第86回日本薬理学会年会, 2013.03.
16. 川田竜, 齊藤 秀俊, 増田 隆博, 津田 誠, 井上 和秀, 転写因子MafBは末梢神経損傷後の神経障害性疼痛発症に寄与する, 第86回日本薬理学会年会, 2013.03.
17. Emika Toyomitsu, Makoto Tsuda, Tomohiro Yamashita, Hidetoshi Tozaki-Saitoh, Yoshitaka Tanaka, Kazuhide Inoue, CCL2 promotes P2X4 receptor trafficking to the cell surface of microglia, International symposium on purinergic signaling, 2012.05, P2X4 receptors, a subtype of the purinergic P2X family, play important roles in regulating neuronal and glial functions in the nervous system. We have previously shown that the expression of P2X4 receptors is upregulated in activated microglia after peripheral nerve injury and that activation of the receptors by extracellular ATP is crucial for maintaining nerve injury-induced pain hypersensitivity. However, the regulation of P2X4 receptor expression on the cell surface of microglia is poorly understood. Here, we identify the CC chemokine receptor CCR2 as a regulator of P2X4 receptor trafficking to the cell surface of microglia. We found that applying CCL2 or CCL12, endogenous ligands for CCR2, to primary cultured microglial cells, increased the levels of P2X4 receptor protein on the cell surface without changing total cellular expression. This effect of CCL2 was prevented by an antagonist of CCR2. Time-lapse imaging of green fluorescent protein-tagged P2X4 receptor in living microglial cells showed that CCL2 stimulation increased the movement of P2X4-GFP particles. The subcellular localization of P2X4 receptor immunofluorescence was restricted to lysosomes around the perinuclear region. Notably, CCL2 changed the distribution of lysosomes with P2X4 receptor immunofluorescence within microglial cells and induced release of the lysosomal enzyme, indicating lysosomal exocytosis. Moreover, CCL2-stimulated microglia enhanced Akt phosphorylation by ATP applied extracellularly, a P2X4 receptor-mediated response. These results indicate that CCL2 promotes expression of P2X4 receptor protein on the cell surface of microglia through exocytosis of P2X4 receptor-containing lysosomes, which may be a possible mechanism for pain hypersensitivity after nerve injury..
18. 米田聡介 増田潤也 齊藤秀俊 津田誠 井上和秀, Role of transcription factor MafB in microglial activation after peripheral nerve injury, 第85回日本薬理学会年会, 2012.03, Neuropathic pain is a chronic pain condition that occurs after nerve damage induced by bone compression in cancer, diabetes, or physical injury. Accumulating evidence has revealed that microglial activation in spinal dorsal horn are involved in neuropathic pain. After nerve injury, microglia in the normal state are converted to an activated state and increase expression of multiple molecules, including cell surface receptors and inflammatory cytokines. However, little is known about the mechanism of microglial activation after nerve injury.
In the present study we examined the involvement of transcription factor MafB in microglia activation after nerve injury. We found that MafB mRNA was increased in spinal cord after peripheral nerve injury (PNI) and that MafB protein was up-regulated specifically in microglia. To elucidate MafB function in microglia, we used lentiviruses encoding shRNAs to knockdown MafB in microglial cell line BV2. Down-regulation of MafB decreased the expression of receptors and inflammatory cytokines. Moreover treatment with several cytokines increased after PNI up-regulated MafB expression in microglia.
These results suggest that MafB upregulated in microglia after PNI is one of the key factors for microglial activation.
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19. 齊藤秀俊、宮田広行、津田誠、井上和秀, 神経障害性疼痛におけるミクログリアP2Y12受容体の関与, 第39回薬物活性シンポジウム, 2011.11, 神経の傷害や機能異常を原因とした神経障害性疼痛では、触刺激を痛みとして感覚してしまうアロディニアと呼ばれる症状が頻発するが、その発症機序には未解明な点が多く、これらを明らかにすることは急務の課題である。現在までに、神経障害性疼痛の発症機序において脊髄ミクログリアの活性化は重要な役割を担っており、さらにはミクログリアに発現する細胞外ヌクレオチドに対する受容体の関与が明らかにされている。P2Y12受容体は細胞外ヌクレオチドの一つであるADPを受容し中枢においてはミクログリアの障害センサーとして働くことが示唆されているが、我々は脊髄ミクログリアのP2Y12受容体が神経因性疼痛の発症機序においても重要な役割を担っていることを報告した。しかしながらその詳細なメカニズムの解明には未だ至ってはいない。
今回我々は、初代培養ミクログリアのP2Y12受容体刺激は一過性の細胞内カルシウム上昇を引き起こし、これによってケモカインCCL3の発現が惹起されることを見出した。CCL3の髄腔内投与は即時に動物の痛み行動を誘起し、この影響は数日間持続するものであった。CCL3中和抗体を神経障害性モデル動物の脊髄に投与すると疼痛形成は有意に抑制された。
以上の結果は、神経障害性疼痛の発症機序でのP2Y12受容体の役割において、ケモカイン発現誘導受容体としての関与を示唆するものである。
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20. Junya Masuda, Tozaki-Saitoh Hidetoshi, Sosuke Yoneda, Makoto Tsuda, Kazuhide Inoue, Transcriptional factor MafB regulates microglial proliferation and gene expression, Neuroscience 2011 , 2011.11, MafB is a transcriptional factor that is highly expressed in monocytes and macrophages in the hematopoietic system, and its expression is strongly upregulated after inducing macrophage differentiation of myeloid progenitor cells. Thus, MafB has potential to determine the macrophage characteristics. In the central nervous system (CNS), microglia, a type of glial cells that play immune roles in the CNS, are recently revealed that the cells are derived from primitive myeloid progenitors. In the pathological states, they undergo activation represented in hypertrophic morphology, proliferation, induction of inflammatory genes, and phagocytic activity. However, the role of MafB in the process of microglial activation is completely unknown. Here we report that MafB expressed in microglia plays crucial roles in regulating cell proliferation and gene expression. We investigated the MafB function by using the lentiviral vector-mediated systems in cultured primary microglia. It was found that overexpression of MafB gene provided high proliferation activity to non-proliferative primary cultured microglia compared to the control groups expressing mutated MafB. Consistent with this observation, microglial cell line BV2 cells that normally show high mitotic activity lost their highly proliferative activity by supression of MafB expression (又はMafB knockdown) using shRNA targeted to MafB (MafB knockdown). These results suggest that MafB controls microglial proliferating activity. In addition, the expression of several genes was varied by MafB expression level. Knockdown of MafB gene in BV2 cells by shRNA-expressing lentivirus leaded to the decreased mRNA expression level of P2 receptors, toll-like receptors, pro-inflammatory cytokines, and transcriptional factors, indicating that MafB broadly regulates functional phenotypes of microglia. Taken together, MafB can be a novel, critical transcriptional regulator of microglia that prominently controls the abilities of their proliferation and gene expression..
21. Hidetoshi Tozaki-Saitoh, Makoto Tsuda, Hiroshi Kiyama, Keiko Ohsawa, Kazuhide Inoue, MicroglialP2Y12receptor as a potential therapeutic target of neuropathic pain, 4thJoint German-Italian PurineClub Meeting, 2011.07, Extracellular nucleotides have been implicated as signaling molecules used by microglia to sense adverse physiological conditions. They act through purinoceptors, especially P2Y12 receptor (P2Y12R) is closely linked to microglial chemotactic function. Emerging evidence has indicated that activated spinal microglia are key cellular intermediaries in the neuropathic pain. However, the role of microglial P2Y12Rs in neuropathic pain remains unclear. Here, we show that the level of P2Y12R expression was markedly increased in the spinal cord, and that this expression was highly restricted to spinal microglia. Mice lacking P2ry12 displayed impaired tactile allodynia after nerve injury, and the intrathecal administration of P2Y12R antagonist prevented the development of tactile allodynia. Using electron microscopy we found an increased number of microglial cells adhering to and engulfing both injured and uninjured myelinated axons after nerve injury. These events were also significantly suppressed by the P2Y12R antagonist. Together, our findings indicate that activation of P2Y12Rs in spinal microglia may be a critical for physical interaction between microglia and neurons, and therefore in the pathogenesis of neuropathic pain..
22. 津田 誠、Knut Biber、齊藤 秀俊、塚本 恵子、豊満 笑加、増田 隆博、Hendrikus Boddeke、井上 和秀, ミクログリアでのP2X4受容体発現を増加させ神経障害性疼痛の形成に関与する損傷ニューロン由来因子CCL21, 第84回日本薬理学会年会, 2011.03.
23. 宮田 広行、齊藤 秀俊、津田 誠、井上 和秀, 初代培養ミクログリアにおけるP2Y12受容体を介したCCL3ケモカインの発現制御, 第84回日本薬理学会年会, 2011.03.
24. 増田 潤哉、齊藤 秀俊、津田 誠、井上 和秀, 神経障害性疼痛モデルマウスにおける活性化型脊髄ミクログリアは高い遊走能を有する, 第84回日本薬理学会年会, 2011.03.
25. 齊藤 秀俊、前田 光代、津田 誠、木山 博資、井上 和秀, 神経障害性疼痛における脊髄ミクログリアP2Y12受容体の関与, 第84回日本薬理学会年会, 2011.03.
26. 片岡 彩子、上杉 歩未、齊藤 秀俊、津田 誠、井上 和秀, UDP刺激誘発性のミクログリア貪食におけるPKDの関与, 第84回日本薬理学会年会, 2011.03.
27. 片岡 彩子、齊藤 秀俊、上杉 歩未、古賀 結衣、津田 誠、井上 和秀, UDPによるミクログリアの貪食促進作用におけるPKDの関与, 第33回日本神経科学大会 第53回日本神経化学会大会 第20回日本神経回路学会大会合同大会, 2010.09.
28. 増田 潤哉、齋藤 秀俊、津田 誠、井上 和秀, 急性脳スライスにおけるミクログリアの状態のリアルタイムイメージング解析, 第33回日本神経科学大会 第53回日本神経化学会大会 第20回日本神経回路学会大会合同大会, 2010.09.
29. Hidetoshi Tozaki-Saitoh, Makoto Tsuda, Hiroyuki Miyata, Kazuaki Ueda, Kazuhide Inoue., P2Y12 receptor in spinal microglia are required for neuropathic pain, International symposium on Purinergic signalling in New strategy of drug discovery, Joint with JSPS Core-to-Core Program, A satellite symposium fro IUPS 2009, 2009.07.
30. 齊藤秀俊、津田誠、井上和秀, 神経因性疼痛における脊髄ミクログリアP2Y12受容体の関与, 日本疼痛学会, 2008.07.
31. 齊藤秀俊、津田誠、井上和秀, 神経因性疼痛発症メカニズムにおける脊髄ミクログリアP2Y12受容体の関与, 日本神経科学会, 2008.07.
32. Hidetoshi Tozaki-Saitoh, Makoto Tsuda, Inoue Kazuhide, P2Y12 receptors in spinal microglia are required for neuropathic pain after peripheral nerve injury, PURINES 2008, 2008.07.
33. Hidetoshi Tozaki-Saitoh, Makoto Tsuda, Hiroyuki Miyata, Kazuhide Inoue, P2Y12 receptors in spinal microglia are required for neuropathic pain after peripheral nerve injury, 日本薬理学会, 2008.03.
34. 齊藤 秀俊、津田 誠、小泉 修一、井上 和秀, P2X4受容体発現制御を介したレチノイン酸シグナルの神経因性疼痛への関与
, 第80回日本薬理学会年会, 2007.03, [URL].
35. Hidetoshi Tozaki-Saitoh, Makoto Tsuda, Kazuhide Inoue, Schuichi Koizumi, Youichi Shinozaki, Yoji Sato, Upregulation of microglial P2X4 receptors by retinoic acid, Neuroscience 2006 , 2006.10, [URL].
学会活動
所属学会名
日本神経化学会
日本薬学会
日本神経科学学会
日本疼痛学会
日本薬理学会
学会大会・会議・シンポジウム等における役割
2018.11.17~2018.11.18, 第35回日本薬学会九州支部大会, 座長(Chairmanship).
2019.03.13~2019.03.13, 第28回神経行動薬理若手研究者の集い, 座長(Chairmanship).
2016.07.14~2016.07.15, 新学術領域「グリアアセンブリ」若手の会, 座長(Chairmanship).
2014.03.21~2014.03.23, 第87回日本薬理学会年会, 座長(Chairmanship).
学術論文等の審査
年度 外国語雑誌査読論文数 日本語雑誌査読論文数 国際会議録査読論文数 国内会議録査読論文数 合計
2019年度      
2018年度      
2017年度      
2016年度      
2015年度      
2011年度      
2009年度      
2008年度      
研究資金
科学研究費補助金の採択状況(文部科学省、日本学術振興会)
2013年度~2018年度, 新学術領域研究, 分担, 脳内ミクログリアによるシナプス制御機構と慢性疼痛.
2011年度~2015年度, 基盤研究(S), 分担, 神経障害性疼痛におけるグリア機能の解明.
2011年度~2013年度, 基盤研究(B), 分担, ヒトP2X4受容体の3次元構造を基盤とした神経障害性疼痛抑制分子の探索.
2006年度~2010年度, 学術創成研究費, 分担, 神経因性疼痛発症メカニズムの解明.
2013年度~2015年度, 基盤研究(C), 代表, 神経障害性疼痛を制御するケモカインシグナルの解明.
2011年度~2012年度, 若手研究(B), 代表, マイクロRNAによるミクログリア活性化制御の解明
.
2009年度~2010年度, 若手研究(B), 代表, P2Y12受容体を介した突起進展によるミクログリア活性化シグナル複合体形成の解明.
2007年度~2008年度, 基盤研究(C), 代表, 神経因性疼痛の発症に関与するP2X4遺伝子発現制御メカニズムの解明.
競争的資金(受託研究を含む)の採択状況
2017年度~2019年度, 日本医療研究開発機構, 分担, 線維筋痛症と中枢性感作に関するトランスレーショナルリサーチ:精神神経免疫学的側面からの病態解明と評価法開発.
寄附金の受入状況
2016年度, GSKジャパン研究助成・「ストレス状況下の情動行動と前頭前野シナプス変化の生体イメージング」・代表.
2014年度, 上原財団H26-研究奨励金・「ミクログリアにおける転写因子 Mafb の機能解析」・代表.

九大関連コンテンツ

pure2017年10月2日から、「九州大学研究者情報」を補完するデータベースとして、Elsevier社の「Pure」による研究業績の公開を開始しました。
 
 
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