Isolation of bone marrow-derived microglia and resident microglia from hypothalamic tissue and comparison of expression of various molecules in chronic PS-loaded and sham-treated mice.
(A) Representative FACS chart in chronic PS-loaded mice with whole body radiation and the number of isolated GFP-CD45low (resident microglia) and GFP+CD45low (bone marrow-derived microglia) from mice with whole body radiation and the radiation with head protection (n = 4−6). Total events in FACS were 20000. Data are expressed as mean ± sem. *P < 0.05 with two-tailed Student’s t-test. (B−E) mRNA expression of chemokine receptors: CCR2 (n = 6−8), CX3CR1 (n = 3−5), CXCR4 (n = 3−5) (B), exiting amino acid transporters: EAAT1 (n = 4−5), EAAT2 (n = 3−4) (C), purinergic P2X receptors: P2X4 (n = 3) and P2X7 (n = 3) and P2Y receptors: P2Y1 (n = 3), P2Y12 (n = 3−4) (D), IL-1β (n = 4−6) and TNF-α (n = 3−5) (E). (F) The length of axis of GFP+Iba-1+ microglia (bone marrow-derived microglia, BMDM) and GFP-Iba-1+ microglia (resident microglia. RM) in chronic PS-loaded and sham mice (n = 4). Scale bars: 10 µm. Data are expressed as mean ± sem. *P < 0.05, **P < 0.01 with ANOVA followed by Tukey’s multiple comparison.