Figure 2.
Figure 2. Double transgenic mice lacking TrkB in GFAP+ cells are protected from EAE. (A) TrkB depletion in astrocytes in GFAP-cre:TrkBflox/flox mice. Immunohistochemistry for β-galactosidase (β-Gal; left), GFAP (middle), and TrkB (right) in the spinal cord of TrkBflox/flox (top) and GFAP-cre:TrkBflox/flox (bottom) naive mice (brown staining with DAB and hematoxylin counterstaining). (B) Immunohistochemistry for GFAP (left) and TrkB (middle) and double immunofluorescence (right) in the spinal cord of GFAP-cre:TrkBflox/flox EAE mice. DAPI was used to stain for nuclei. Arrows indicate TrkB-positive immune cells. (A and B) The antibody directed to the ECD of TrkB (Cell Signaling Technology) was used. Bars, 50 µm. (C) Quantification of TrkB-T1 immunoreactivity in spinal cord white matter of naive and EAE mice. AU, arbitrary units. (D) Colocalization between TrkB-T1 and GFAP in naive and EAE mice. Data are given as mean percentage of GFAP signal. (C and D) Four mice/group from two independent EAE experiments and two naive mice/group were analyzed. Error bars represent SEM. (E) EAE disease course. Clinical expression of MOG35–55-induced EAE in GFAP-cre:TrkBflox/flox mice (n = 13) compared with TrkBflox/flox control littermates (n = 20). P-value lines refer to significant differences over time. Error bars indicate SEM. (F) Table summarizing EAE clinical parameters in GFAP-cre:TrkBflox/flox mice and control littermates. Data from three EAE experiments were pooled, analyzed, and given as mean ± SEM. Only diseased animals were included in the analysis for the onset, max score, score at peak, and cumulative disease index (C.D.I.). p.i., postimmunization. (G) Proliferation of draining lymph node (drLNs; top) or spleen (bottom) cells of GFAP-cre:TrkBflox/flox (n = 5) and TrkBflox/flox (n = 5) mice at day 10 after immunization with MOG35–55 peptide, as assessed by [3H]thymidine incorporation. Values are expressed as percentage of unstimulated control cells. Similar observations were obtained in a second experiment. Error bars indicate SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Double transgenic mice lacking TrkB in GFAP+ cells are protected from EAE. (A) TrkB depletion in astrocytes in GFAP-cre:TrkBflox/flox mice. Immunohistochemistry for β-galactosidase (β-Gal; left), GFAP (middle), and TrkB (right) in the spinal cord of TrkBflox/flox (top) and GFAP-cre:TrkBflox/flox (bottom) naive mice (brown staining with DAB and hematoxylin counterstaining). (B) Immunohistochemistry for GFAP (left) and TrkB (middle) and double immunofluorescence (right) in the spinal cord of GFAP-cre:TrkBflox/flox EAE mice. DAPI was used to stain for nuclei. Arrows indicate TrkB-positive immune cells. (A and B) The antibody directed to the ECD of TrkB (Cell Signaling Technology) was used. Bars, 50 µm. (C) Quantification of TrkB-T1 immunoreactivity in spinal cord white matter of naive and EAE mice. AU, arbitrary units. (D) Colocalization between TrkB-T1 and GFAP in naive and EAE mice. Data are given as mean percentage of GFAP signal. (C and D) Four mice/group from two independent EAE experiments and two naive mice/group were analyzed. Error bars represent SEM. (E) EAE disease course. Clinical expression of MOG35–55-induced EAE in GFAP-cre:TrkBflox/flox mice (n = 13) compared with TrkBflox/flox control littermates (n = 20). P-value lines refer to significant differences over time. Error bars indicate SEM. (F) Table summarizing EAE clinical parameters in GFAP-cre:TrkBflox/flox mice and control littermates. Data from three EAE experiments were pooled, analyzed, and given as mean ± SEM. Only diseased animals were included in the analysis for the onset, max score, score at peak, and cumulative disease index (C.D.I.). p.i., postimmunization. (G) Proliferation of draining lymph node (drLNs; top) or spleen (bottom) cells of GFAP-cre:TrkBflox/flox (n = 5) and TrkBflox/flox (n = 5) mice at day 10 after immunization with MOG35–55 peptide, as assessed by [3H]thymidine incorporation. Values are expressed as percentage of unstimulated control cells. Similar observations were obtained in a second experiment. Error bars indicate SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001.

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