Journal: PLoS ONE

Article Title: Aberrant Immune Responses in a Mouse with Behavioral Disorders

doi: 10.1371/journal.pone.0020912

Figure Lengend Snippet: Sera from postnatal day 21 mice were added to wells coated with BALB/c SCID whole brain proteins (10 µg/well). The numbers of sera were 6 for BTBR male or female, 6 for BCF1 male, 3 for BCF1 female, 5 for CBF1 male or female, and 7 for B6 male or female. The level of brain-reactive IgG was determined by using the HRP-conjugated goat anti-mouse IgG. *, p <0.01 vs. B6 males or females, and **, p <0.05 vs. B6 males.

Article Snippet: The level of total IgG in serum or brain homogenates was determined by a sandwich ELISA using goat anti-mouse IgG Fc (Pierce, Rockford, IL) as a capture Ab and HRP-goat anti-mouse IgG as a detection Ab (Sigma).

Techniques:

Journal: Glia

Article Title: CHPG Enhances BDNF and Myelination in Cuprizone-Treated Mice through Astrocytic Metabotropic Glutamate Receptor 5

doi: 10.1002/glia.24003

Figure Lengend Snippet: Western blots demonstrated BDNF, MBP, and PLP protein levels in the corpus callosum of mice subjected to a 4-week cuprizone (Cupz) lesion and injected intraperitoneally with CHPG (40 mg/kg) or 0.9% saline (Sal) vehicle 6 and 24 hours prior to dissection. Graph represents densitometric analysis of Western blots normalized to GAPDH and presented as percent saline-injected control-fed (Ctrl) mice, analyzed by ANOVA followed by Tukey’s multiple comparisons test. *Significantly different from saline-injected and CHPG-injected control-fed mice, *p<0.05, #Significantly different from saline-injected cuprizone-fed mice, #p<0.05. Each experiment consisted of one mouse per treatment group and each experiment was repeated four times. Data are presented as mean ± SEM.

Article Snippet: Antibodies for Western blots included a rabbit polyclonal antibody to BDNF (1:200, Cat# sc-546, Santa Cruz Biotechnology, RRID:AB_630940), a mouse monoclonal antibody to myelin basic protein (MBP) (1:400, Cat# MCA184S, Serotec, RRID:AB_322319), and a goat polyclonal antibody to PLP (1:1,000, Cat# sc-18529, Santa Cruz Biotechnology, RRID:AB_2165798).

Techniques: Western Blot, Injection, Saline, Dissection, Control

Journal: Glia

Article Title: CHPG Enhances BDNF and Myelination in Cuprizone-Treated Mice through Astrocytic Metabotropic Glutamate Receptor 5

doi: 10.1002/glia.24003

Figure Lengend Snippet: A) Western blots demonstrated BDNF, MBP, and PLP protein levels in the corpus callosum of hGFAP-CreERT2-BDNFfl/fl-eGFP mice (Cre+) or BDNFfl/fl-eGFP mice lacking cre expression (Cre−) subjected to a 4-week cuprizone (Cupz) lesion and injected intraperitoneally with CHPG (40 mg/kg) or 0.9% saline (Sal) vehicle 6 and 24 hours prior to dissection. Graph represents densitometric analysis of Western blots normalized to GAPDH and presented as percent saline-injected cuprizone-fed mice. *Significantly different, *p<0.05, analyzed by ANOVA followed by Tukey’s multiple comparisons test. Each experiment consisted of one mouse per treatment group and each experiment was repeated four times. Data are presented as mean ± SEM. B) Western blots demonstrated BDNF, MBP, and PLP protein levels in the corpus callosum of PLP-CreERT2-TrkBfl/fl-eGFP mice (Cre+) or TrkBfl/fl-eGFP mice lacking cre expression (Cre−) subjected to a 4-week cuprizone (Cupz) lesion and injected intraperitoneally with CHPG (40 mg/kg) or 0.9% saline (Sal) vehicle 6 and 24 hours prior to dissection. Graph represents densitometric analysis of Western blots normalized to GAPDH and presented as percent saline-injected cuprizone-fed mice. *Significantly different, *p<0.05, analyzed by ANOVA followed by Tukey’s multiple comparisons test. Each experiment consisted of one mouse per treatment group and each experiment was repeated four times. Data are presented as mean ± SEM.

Article Snippet: Antibodies for Western blots included a rabbit polyclonal antibody to BDNF (1:200, Cat# sc-546, Santa Cruz Biotechnology, RRID:AB_630940), a mouse monoclonal antibody to myelin basic protein (MBP) (1:400, Cat# MCA184S, Serotec, RRID:AB_322319), and a goat polyclonal antibody to PLP (1:1,000, Cat# sc-18529, Santa Cruz Biotechnology, RRID:AB_2165798).

Techniques: Western Blot, Expressing, Injection, Saline, Dissection

Journal: Glia

Article Title: CHPG Enhances BDNF and Myelination in Cuprizone-Treated Mice through Astrocytic Metabotropic Glutamate Receptor 5

doi: 10.1002/glia.24003

Figure Lengend Snippet: A) Western blots demonstrated BDNF, MBP, and PLP protein levels in the corpus callosum of mice subjected to a 4-week cuprizone (Cupz) lesion and injected intraperitoneally with CHPG (40 mg/kg) or 0.9% saline (Sal) vehicle 1 hour following stereotaxic injection of the mGluR5 antagonist MPEP (150 μM) or saline. Mice were dissected 24 hours after CHPG injection. Graph represents densitometric analysis of Western blots normalized to GAPDH and presented as percent saline-injected cuprizone-fed mice. *Significantly different, *p<0.05, analyzed by ANOVA followed by Tukey’s multiple comparisons test. Each experiment consisted of one mouse per treatment group and each experiment was repeated five times. Data are presented as mean ± SEM. B) mGluR5+GFAP+ immunofluorescent staining in the corpus callosum of hGFAP-CreERT2-mGluR5fl/fl-eGFP mice (Cre+) or mGluR5fl/fl-eGFP mice lacking cre expression (Cre−) subjected to a 4-week cuprizone lesion. Data presented as percent mGluR5+GFAP+ cellular profiles per total GFAP+ cellular profiles relative to Cre− mice. *Significantly different from Cre− cuprizone control, *p<0.05, analyzed by paired t-test. Each experiment consisted of one mouse per treatment group and each experiment was repeated three times. Data are presented as mean ± SEM. Scale bar, 20 μm. C) Western blots demonstrated BDNF, MBP, and PLP protein levels in the corpus callosum of hGFAP-CreERT2-mGluR5fl/fl-eGFP mice (Cre+) or mGluR5fl/fl-eGFP mice lacking cre expression (Cre−) subjected to a 4-week cuprizone (Cupz) lesion and injected intraperitoneally with CHPG (40 mg/kg) or 0.9% saline (Sal) vehicle. Mice were dissected 24 hours after CHPG injection. Graph represents densitometric analysis of Western blots normalized to GAPDH and presented as percent saline-injected cuprizone-fed mice. *Significantly different, *p<0.05, analyzed by ANOVA followed by Tukey’s multiple comparisons test. Each experiment consisted of one mouse per treatment group and each experiment was repeated five times. Data are presented as mean ± SEM.

Article Snippet: Antibodies for Western blots included a rabbit polyclonal antibody to BDNF (1:200, Cat# sc-546, Santa Cruz Biotechnology, RRID:AB_630940), a mouse monoclonal antibody to myelin basic protein (MBP) (1:400, Cat# MCA184S, Serotec, RRID:AB_322319), and a goat polyclonal antibody to PLP (1:1,000, Cat# sc-18529, Santa Cruz Biotechnology, RRID:AB_2165798).

Techniques: Western Blot, Injection, Saline, Staining, Expressing, Control

Journal: Glia

Article Title: CHPG Enhances BDNF and Myelination in Cuprizone-Treated Mice through Astrocytic Metabotropic Glutamate Receptor 5

doi: 10.1002/glia.24003

Figure Lengend Snippet: A) Western blots demonstrated BDNF, MBP, and PLP protein levels in the corpus callosum of mice subjected to a 4-week cuprizone (Cupz) lesion and injected intraperitoneally with CHPG (20 mg/kg) or 0.9% saline (Sal) vehicle every other day for an additional 2 weeks while remaining on cuprizone feed. Graph represents densitometric analysis of Western blots normalized to GAPDH and presented as percent saline-injected control-fed (Ctrl) mice, analyzed by ANOVA followed by Tukey’s multiple comparisons test. *Significantly different from saline-injected and CHPG-injected control-fed mice, *p<0.05, #Significantly different from saline-injected cuprizone-fed mice, #p<0.05. Each experiment consisted of two control mice ± CHPG and two or four cuprizone mice ± CHPG. Each experiment was repeated eight times. Data are presented as mean ± SEM. B) CC1 immunofluorescent staining in the corpus callosum of mice subjected to a 4-week cuprizone (Cupz) lesion and injected intraperitoneally with CHPG (20 mg/kg) or 0.9% saline (Sal) vehicle every other day for an additional 2 weeks while remaining on cuprizone feed. Data presented as percent saline-injected cuprizone-fed (Cupz) mice, analyzed by paired t-test. *Significantly different from saline-injected cuprizone-fed mice, *p<0.05. Each experiment consisted of one mouse per treatment group and each experiment was repeated six times. Data are presented as mean ± SEM. Scale bar, 20 μm.

Article Snippet: Antibodies for Western blots included a rabbit polyclonal antibody to BDNF (1:200, Cat# sc-546, Santa Cruz Biotechnology, RRID:AB_630940), a mouse monoclonal antibody to myelin basic protein (MBP) (1:400, Cat# MCA184S, Serotec, RRID:AB_322319), and a goat polyclonal antibody to PLP (1:1,000, Cat# sc-18529, Santa Cruz Biotechnology, RRID:AB_2165798).

Techniques: Western Blot, Injection, Saline, Control, Staining

Journal: Acta neuropathologica communications

Article Title: BDNF augmentation reverses cranial radiation therapy-induced cognitive decline and neurodegenerative consequences.

doi: 10.1186/s40478-024-01906-9

Figure Lengend Snippet: Fig. 2 Riluzole treatment restores hippocampal BDNF in the irradiated hippocampus. 10–12 weeks old WT male mice received cranial radiation therapy (RT) followed by riluzole (RZ) treatment (13 mg/kg) in drinking water for 6–7 weeks. An ELISA-based quantification of BDNF from the micro-dissected hippocampus showed RT-induced reductions in the RT + Vehicle group. Importantly, RZ treatment in the cranially irradiated mice showed significant restoration of BDNF levels. Data are presented as mean ± SEM (N = 6–10 mice per group). P values were derived from two-way ANOVA and Bonferroni’s multiple comparisons test

Article Snippet: BDNF levels were quantified using a commercially available ELISA kit (E-EL-M0203, Elabscience Biotechnology) and uncoated ELISA plates (Nunc MaxiSorp, Biolegend).

Techniques: Irradiation, Enzyme-linked Immunosorbent Assay, Derivative Assay