A novel optical assay system for the quantitative measurement of chemotaxis

Kanegasaki, Shiro; Nomura, Yujiro; Nitta, Nao; Akiyama, Shuichi; Tamatani, Takuya; Goshoh, Yasuhiro; Yoshida, Takashi; Sato, Tsuyoshi; Kikuchi, Yuji

doi:10.1016/j.jim.2003.07.008

Cited by 113 publications

(115 citation statements)

References 12 publications

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“…To address whether IRF8 is involved in microglial cell motility, we initially placed primary cultured microglial cells from wild-type (Irf8 +/+ ) or IRF8-deficient (Irf8 −/− ) mice in an EZTaxiscan chamber, which generated a concentration gradient of chemoattractants (e.g., ATP and ADP) and assessed chemotactic activity [20,21]. As previously reported [12,22], Irf8 +/+ microglia showed clear and robust chemotactic activity and moved toward the top edge of the chamber where higher concentrations of ATP were located (Fig.…”

Section: Irf8 Deficiency Affects Nucleotide-induced Chemotactic Activmentioning

confidence: 99%

IRF8 is a transcriptional determinant for microglial motility

Masuda

Nishimoto

Tomiyama

et al. 2014

Purinergic Signalling

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Microglia, the resident immune cells of the central nervous system, are constitutively mobile cells that undergo rapid directional movement toward sites of tissue disruption. However, transcriptional regulatory mechanisms of microglial motility remain unknown. In the present study, we show that interferon regulatory factor-8 (IRF8) regulates microglial motility. We found that ATP and complement component, C5a, induced chemotaxis of IRF8 wild-type microglia. However, these responses were markedly suppressed in microglia lacking IRF8 (Irf8 −/− ). In a consistent manner, phosphorylation of Akt (which plays a crucial role in ATP-induced chemotaxis) was abolished in Irf8 −/− microglia. Real-time polymerase chain reaction analysis revealed that motility-related microglial genes such as P2Y 12 receptor were significantly suppressed in Irf8 −/− microglia. Furthermore, Irf8 −/− microglia exhibited a differential expression pattern of nucleotidedegrading enzymes compared with their wild-type counterparts. Overall, our findings suggest that IRF8 may regulate microglial motility via the control of microglial gene expression.

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Section: Irf8 Deficiency Affects Nucleotide-induced Chemotactic Activmentioning

confidence: 99%

IRF8 is a transcriptional determinant for microglial motility

Masuda

Nishimoto

Tomiyama

et al. 2014

Purinergic Signalling

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“…The microchemotaxis chamber (TAXIScan; Effector Cell Institute) was used to detect real-time horizontal chemotaxis according to the method described previously (34). After serum starvation for 3 h, mature DCs (1 ϫ 10 3 cells/ l) and S1P (10 M) were put into the space in each side of the chamber and incubated for 2 h at 37°C.…”

Section: Real-time Chemotaxis Assaymentioning

confidence: 99%

Migration of CD4 T Cells and Dendritic Cells toward Sphingosine 1-Phosphate (S1P) Is Mediated by Different Receptor Subtypes: S1P Regulates the Functions of Murine Mature Dendritic Cells via S1P Receptor Type 3

Maeda¹,

Matsuyuki²,

Shimano³

et al. 2007

The Journal of Immunology

119

105

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Dendritic cells (DCs) and lymphocytes are known to show a migratory response to the phospholipid mediator, sphingosine 1-phosphate (S1P). However, it is unclear whether the same S1P receptor subtype mediates the migration of lymphocytes and DCs toward S1P. In this study, we investigated the involvement of S1P receptor subtypes in S1P-induced migration of CD4 T cells and bone marrow-derived DCs in mice. A potent S1P receptor agonist, the (S)-enantiomer of FTY720-phosphate [(S)-FTY720-P], at 0.1 nM or higher and a selective S1P receptor type 1 (S1P1) agonist, SEW2871, at 0.1 μM or higher induced a dose-dependent down-regulation of S1P1. The pretreatment with these compounds resulted in a significant inhibition of mouse CD4 T cell migration toward S1P. Thus, it is revealed that CD4 T cell migration toward S1P is highly dependent on S1P1. Mature DCs, when compared with CD4 T cells or immature DCs, expressed a relatively higher level of S1P3 mRNA. S1P at 10–1000 nM induced a marked migration and significantly enhanced the endocytosis of FITC-dextran in mature but not immature DCs. Pretreatment with (S)-FTY720-P at 0.1 μM or higher resulted in a significant inhibition of S1P-induced migration and endocytosis in mature DCs, whereas SEW2871 up to 100 μM did not show any clear effect. Moreover, we found that S1P-induced migration and endocytosis were at an extremely low level in mature DCs prepared from S1P3-knockout mice. These results indicate that S1P regulates migration and endocytosis of murine mature DCs via S1P3 but not S1P1.

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“…It also attracts eosinophils, basophils, and a subpopulation of lymphocytes. 17 HNPs and certain chemokines share characteristics such as size, cationic charge, and disulfide bonding and biologic activities such as chemotaxis and antimicrobial properties. 18,19 Although they lack amino acid sequence homology, the 3-dimensional structures of human ␤-defensins 1 and 2 and the CC chemokine CCL20 are similar, and all 3 molecules specifically interact with the CCL20 receptor CCR6 and induce similar biologic responses.…”

Section: Introductionmentioning

confidence: 99%