Figure 6
Immunohistochemistry of GLT1 with Iba1, mRNA expression of GLT1 and glutamate contents in the of SOD1(G93A) mice after treatments. (a) Immunohistochemical analysis of GLT1 (red) with YFP (green) signals at ventral horn in the spinal cord of SOD1(G93A) mice at 10 weeks after bone marrow transplantation with gene transduction by LV-CD68-YFP (upper row panels) or LV-CD68-GLT1-YFP (lower row panels). Left panels show YFP signals, middle panels show GLT1 staining, and right panels show merged images of left and middle panels in the same row. Arrowheads indicate YFP and GLT1 double-positive cells. Arrows indicate GLT1 single-positive cells. Scale bar = 50 µm. (b) GLT1-positive area intensity in SOD1(G93A) mice (n = 6) and SOD1(G93A) mice with the empty vector [SOD1(G93A) + LV-CD68-YFP, n = 6] or treatment vector [SOD1(G93A) + LV-CD68-GLT1-YFP, n = 6]. Error bars represent the mean + SD. *p < 0.05 between the treatment group and others. (c) Immunohistochemical analysis of Iba1 (red) and GLT1 (blue) with YFP (green) signals at ventral horn in the spinal cord of SOD1(G93A) mice at 10 weeks after bone marrow transplantation (18-week-old) with gene transduction by LV-CD68-GLT1-YFP. The most left panel shows YFP signals (green), the second, third and fourth panel from the left side show Iba1 (red), GLT1 (blue) staining and the merged images of three left side panels. Arrowheads indicate YFP, Iba1 and GLT1 triple-positive cells. Arrows indicate GLT1 positive cells without YFP signals. Scale bar = 50 µm. (d) mRNA expression level of GLT1 in SOD1(G93A) mice (n = 6) and SOD1(G93A) mice with the empty vector [SOD1(G93A) + LV-CD68-YFP, n = 6] or treatment vector [SOD1(G93A) + LV-CD68-GLT1-YFP n = 6]. Data were normalized to GAPDH mRNA expression. (e) Glutamate concentration in cerebrospinal fluid in SOD1(G93A) mice, SOD1(G93A) mice with the empty vector [SOD1(G93A) + LV-CD68-YFP] or treatment vector [SOD1(G93A) + LV-CD68-GLT1-YFP] (n = 4 in each group). Error bars represent the mean + SD. *p < 0.05 between the treatment group and others. CSF cerebrospinal fluid.
