Figure 2

PDGF-BB phosphorylates its receptors in spinalmicroglia. (A) The immunoreactivity of phosphorylated PDGFRβ protein was detected by a specific antibody for p-PDGFRβ 30 min after intrathecal administration of vehicle or PDGF-BB (10 pmol) in the L5 spinal dorsal horn. Scale bar, 200 μm. (B) The intensity of p-PDGFRβ immunofluorescence was quantified in the dorsal horn region of vehicle treated rats and PDGF-BB treated rats. Data represent the means ± SEM of the immunofluorescence intensity (n = 5). ***P < 0.001 vs vehicle by Student's t-test. (C) Double immunofluorescence labeling of the dorsal horn 30 min after intrathecal PDGF-BB administration with p-PDGFRβ (green) and cell markers (red); OX-42, a microglia marker; GFAP, an astrocytes marker; CC1, an oligodendrocytes marker; NeuN and MAP2, neurons markers. Scale bars, 20 μm. (D) PDGFRα (116 bp) and PDGFRβ (145 bp) mRNA expression in primary microglia by RT-PCR analysis. Spinal cord, cerebral cortex, and spleen are positive controls. (E) Triple immunofluorescence labeling of p-PDGFRβ (green) with OX-42 (red) and DAPI (blue), a nuclear marker, in primary microglia treated with PBS as a control or PDGF-BB (50 ng/ml) for 10 min. Scale bar, 20 μm.