N from chronic PSloaded and sham-treated mice. Arrows indicate MCP-1+NeuN
N from chronic PSloaded and sham-treated mice. Arrows indicate MCP-1+NeuN+ cells and arrow heads indicate MCP-1-NeuN+ cells. Scale bars: 20 . Information are expressed as mean sem (n = 7). *P 0.05 with two-tailed Student’s t-test. (C) Immunofluorescence staining with MCP-1 (red) and GFAP (pink) in PVN from chronic PS-loaded and sham-treated mice. Scale bars: 20 . Data are expressed as mean sem (n = 4-6). *P 0.05 with two-tailed Student’s t-test. (D) Frequency of GFP+CCR2+ cells inside the peripheral blood obtained by FACS (n = 4-6). Data are expressed as mean sem. *P 0.05 with two-tailed Student’s t-test. (E) The number of CCR2+ cells within the hypothalamus obtained by FACS (n = four). Total events in FACS have been 20000. Data are expressed as mean sem. *P 0.05 with ANOVA followed by Tukey’s several comparison. (F) eNOS Compound Effects of a CCR2 antagonist, RS102895, on infiltration of bone marrow-derived microglia in PVN. PVN is shown by dotted line. Scale bars: 50 . Variety of GFP-positive cells within a section such as PVN is shown. (G) Effect of RS102895 around the anxiety-like behavior induced by chronic PS. Data are expressed as mean sem. *P 0.05 with ANOVA followed by Tukey’s a number of comparison (n = 4).doi: ten.1371/journal.pone.0081744.gand not received bone marrow transplantation, the MCP-1/ CCR2 axis in the brain was JAK3 Species involved in the mechanism of the abnormal behavior induced by chronic PS. The recruitment of bone marrow cells into the peripheral circulation is regulated by the SDF-1/CXCR4 interaction amongst bone marrow niche cells and hematopoietic stem cells [302]. A current study showed that circadian oscillation originating in the suprachiasmatic nucleus within the brainstimulates 3-adrenergic receptors on bone marrow stromal cells by means of sympathetic pathways [32]. Activation of 3-adrenergic receptors on bone marrow stromal cells induces the reduction of SDF-1 production inside the stromal cells, resulting in the decrease in the concentration of SDF-1 in the bone marrow. Within the specific microenvironments, niche, some stromal cells keep the proliferation and differentiation of hematopoietic stem cells, which express CXCR4 on their surfaces. ThePLOS 1 | plosone.orgChronic Stress and Bone Marrow-Derived MicrogliaFigure 4. Expression of SDF-1 within the bone marrow and frequency of GFP+CXCR4+ cells in peripheral blood. Effects of 3adrenergic blockade on the infiltration of bone marrow-derived microglia in the PVN. (A) Chronic PS decreased the SDF-1 concentration within the bone marrow (n = 3-4) and elevated the frequency of CXCR4+ monocytes inside the peripheral blood (n = 4-6). (B, C) 3-adrenoceptor antagonist SR59230A blocks the infiltration of bone marrow-derived microglia in the PVN. Variety of GFPpositive cells was counted within a section including PVN. Data are expressed as imply sem. *P 0.05 (n = 3-11). PVN shown by dotted line. Scale bars: 50 .doi: 10.1371/journal.pone.0081744.gPLOS A single | plosone.orgChronic Stress and Bone Marrow-Derived MicrogliaFigure five. 3 dimensional image of immunofluorescence staining of IL-1, pNMDA receptor, and IL-1 receptor in PVN from chronic PS-loaded mouse. (A) GFP-positive cells (green) overlapped with IL-1 (red). (B, C) Good reactions for pNMDAR (pink) and IL-1R (pink) detected around the membrane of neurons (red) adjacent to bone marrow-derived microglia (green) in PVN. Scale bars: 10 .doi: 10.1371/journal.pone.0081744.gSDF-1/CXCR4 interaction is an important axis for the hematopoiesis. For that reason, the circadian oscillation stimulates.