Journal: Neurochemical Research

Article Title: Reactive Astrocytes Release GDNF to Promote Brain Recovery and Neuronal Survival Following Ischemic Stroke

doi: 10.1007/s11064-025-04370-6

Figure Lengend Snippet: The effects of GDNF overexpression in astrocytes on synaptic proteins following PT. (a) An illustration of the viral injection. IC: ischemic core; PIR: peri-infarct region. (b) Experimental timeline. (c , d) Fluorescent images showing the expressions of GDNF and mRFP in cortical astrocytes 7 days after PT. (e-l) WB images (e) and summary data (f-l) of protein expressions in cortical tissues 7 days after PT. In (c-l) rAAV5-GDNF and rAAV5-mRFP virus were injected 2 weeks before PT. N = 4 mice for each group. * p < 0.05, ** p < 0.01, *** p < 0.001; (Student’s t-test)

Article Snippet: The primary antibodies include a mouse anti-GFAP monoclonal antibody (1:300; Cat. No. MAB360, Sigma), a rabbit anti-GDNF polyclonal antibody (1:300, Cat. No. SC-328, Santa Cruz Biotechnology), a rabbit anti-Iba 1 polyclonal antibody (1:300, Cat. No. 019-19741, Fujifilm).

Techniques: Over Expression, Injection, Virus

Journal: Neurochemical Research

Article Title: Reactive Astrocytes Release GDNF to Promote Brain Recovery and Neuronal Survival Following Ischemic Stroke

doi: 10.1007/s11064-025-04370-6

Figure Lengend Snippet: Overexpression of GDNF in astrocytes reduced brain lesion and promoted motor function recovery after PT. (a) Representative images of Nissl staining showing the brain infarct areas of viral injected mice 2 days after PT. The white dash lines outline the damage area. (b , c) Quantification of infarct volumes at 2 (b) and 7 (c) days after PT. N = 4 mice for each group 2 days after PT and 7 mice for each group 7 days after PT. (d-g) Evaluation of motor behavior function by cylinder (d , e) , hanging wire (f) , and grip force (g) tests at different timepoints before and after PT. N = 8 and 7 mice injected with rAAV5-mRFP and rAAV5-GDNF vectors. * p < 0.05, ** p < 0.01, ***<0.001; (Student’s t-test)

Article Snippet: The primary antibodies include a mouse anti-GFAP monoclonal antibody (1:300; Cat. No. MAB360, Sigma), a rabbit anti-GDNF polyclonal antibody (1:300, Cat. No. SC-328, Santa Cruz Biotechnology), a rabbit anti-Iba 1 polyclonal antibody (1:300, Cat. No. 019-19741, Fujifilm).

Techniques: Over Expression, Staining, Injection

Journal: Neurochemical Research

Article Title: Reactive Astrocytes Release GDNF to Promote Brain Recovery and Neuronal Survival Following Ischemic Stroke

doi: 10.1007/s11064-025-04370-6

Figure Lengend Snippet: GDNF overexpression promoted proliferation of reactive astrocytes and reduced oxidative stress in PIR following PI. (a-d) Representative fluorescent images and signal analysis of GFAP (a , b) and Iba 1 (c , d) expressions in the PIR from rAAV5-mRFP and rAAV5-GDNF viruses injected mice at day 4 post stroke. The right panels were the high-resolution images of the boxed region in the left panels. N = 4 mice for each group. (e-f) Fluorescent images of DHE and Dapi and analysis of DHE signal in the PIR of mRFP and GDNF viruses transduced mice at 4 days after stroke. The right panels were the high-resolution images of the boxed region in the left panels. Notice the decreases in DHE signal in GDNF overexpression mice. Data were averaged from 10 images for each group. * p < 0.05, ** p < 0.01, *** p < 0.001; (Student’s t-test)

Article Snippet: The primary antibodies include a mouse anti-GFAP monoclonal antibody (1:300; Cat. No. MAB360, Sigma), a rabbit anti-GDNF polyclonal antibody (1:300, Cat. No. SC-328, Santa Cruz Biotechnology), a rabbit anti-Iba 1 polyclonal antibody (1:300, Cat. No. 019-19741, Fujifilm).

Techniques: Over Expression, Injection

Journal: Neurochemical Research

Article Title: Reactive Astrocytes Release GDNF to Promote Brain Recovery and Neuronal Survival Following Ischemic Stroke

doi: 10.1007/s11064-025-04370-6

Figure Lengend Snippet: Astrocyte-derived GDNF reduced neuronal death after OGD. (a , b) WB images and analysis of GDNF expression in primary cultured astrocytes (a) and GDNF content in the medium (b) at different conditions. N = 4 independent experiments. For OGD condition, primary cultured astrocytes were subjected to 6 h OGD and followed by 24 h reperfusion for western blot and ELISA analysis. (c) Schematic diagram showing experimental design. Primary astrocytes with or without transfection of GDNF plasmid were subjected to 6 h OGD. Astrocyte conditioned medium from astrocyte cultures not subjected or subjected to OGD, namely ACM and ACM(OGD) of different conditions were collected 24 h later and added to primary neuronal cultures after subjected to 1 h OGD followed by 24 h reoxygenation for neuronal viability and death assays. (d-f) Neuronal viability, percentages of PI + neurons and neurons with condensed nuclei after 1 h OGD and followed by 24 h reoxygenation at different conditions. The percentage of PI + neurons was calculated based on the total number of neurons based on DAPI staining. (g) Fluorescent images of neurons stained with PI and Dapi at different conditions. Data in (d) were averaged values from 6 replicates of representative experiment; data in (e , f) were averaged from cell counting of 9 images in each condition. * p < 0.05, ** p < 0.01, *** p < 0.001; (one-way ANOVA test)

Article Snippet: The primary antibodies include a mouse anti-GFAP monoclonal antibody (1:300; Cat. No. MAB360, Sigma), a rabbit anti-GDNF polyclonal antibody (1:300, Cat. No. SC-328, Santa Cruz Biotechnology), a rabbit anti-Iba 1 polyclonal antibody (1:300, Cat. No. 019-19741, Fujifilm).

Techniques: Derivative Assay, Expressing, Cell Culture, Western Blot, Enzyme-linked Immunosorbent Assay, Transfection, Plasmid Preparation, Staining, Cell Counting

Journal: Neurochemical Research

Article Title: Reactive Astrocytes Release GDNF to Promote Brain Recovery and Neuronal Survival Following Ischemic Stroke

doi: 10.1007/s11064-025-04370-6

Figure Lengend Snippet: Reactive astrocytes-derived GDNF triggered the activation of RET receptors in primary neurons after OGD. (a-e) WB images of protein expressions (a) and analysis (b-e) in primary cultured neurons after OGD. Summary data in (b-e) were averaged from 4 replicates. * p < 0.05, ** p < 0.01; (one-way ANOVA test)

Article Snippet: The primary antibodies include a mouse anti-GFAP monoclonal antibody (1:300; Cat. No. MAB360, Sigma), a rabbit anti-GDNF polyclonal antibody (1:300, Cat. No. SC-328, Santa Cruz Biotechnology), a rabbit anti-Iba 1 polyclonal antibody (1:300, Cat. No. 019-19741, Fujifilm).

Techniques: Derivative Assay, Activation Assay, Cell Culture

Journal: Neurochemical Research

Article Title: Reactive Astrocytes Release GDNF to Promote Brain Recovery and Neuronal Survival Following Ischemic Stroke

doi: 10.1007/s11064-025-04370-6

Figure Lengend Snippet: Astrocyte-derived GDNF protected neurons against OGD through the activation of neuronal GDNF receptors. (a-c) Neuronal viability and death based on PI staining and condensed nuclei after 1 h OGD and followed by 24 h reoxygenation at different conditions. (d) Representative fluorescent images of neurons stained with PI and Dapi at different conditions. Data in (a) were averaged values for 6 replicates of representative experiment; data in (b , c) were averaged from cell counting in 9 images in each condition. * p < 0.05, ** p < 0.01, *** p < 0.001; (one-way ANOVA test)

Article Snippet: The primary antibodies include a mouse anti-GFAP monoclonal antibody (1:300; Cat. No. MAB360, Sigma), a rabbit anti-GDNF polyclonal antibody (1:300, Cat. No. SC-328, Santa Cruz Biotechnology), a rabbit anti-Iba 1 polyclonal antibody (1:300, Cat. No. 019-19741, Fujifilm).

Techniques: Derivative Assay, Activation Assay, Staining, Cell Counting

Journal: Neurochemical Research

Article Title: Reactive Astrocytes Release GDNF to Promote Brain Recovery and Neuronal Survival Following Ischemic Stroke

doi: 10.1007/s11064-025-04370-6

Figure Lengend Snippet: Reactive astrocyte-derived GDNF inhibited mitochondrial fission and apoptosis of primary neurons after OGD. (a-g) WB images (a) and analysis (b-g) of protein expressions in primary neurons after OGD. Data were from 3–4 independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001; (one-way ANOVA test)

Article Snippet: The primary antibodies include a mouse anti-GFAP monoclonal antibody (1:300; Cat. No. MAB360, Sigma), a rabbit anti-GDNF polyclonal antibody (1:300, Cat. No. SC-328, Santa Cruz Biotechnology), a rabbit anti-Iba 1 polyclonal antibody (1:300, Cat. No. 019-19741, Fujifilm).

Techniques: Derivative Assay

Journal: Neurochemical Research

Article Title: Reactive Astrocytes Release GDNF to Promote Brain Recovery and Neuronal Survival Following Ischemic Stroke

doi: 10.1007/s11064-025-04370-6

Figure Lengend Snippet: Schematic diagram illustrating the role of astrocytic GDNF in neuronal survival and brain recovery after ischemic stroke. Astrocytes are activated following ischemic stroke and become reactive astrocytes. Reactive astrocytes release GDNF into extracellular space and binds to GFRα1 protein anchor on neuronal cell membrane. The GDNF-GFRα1 complexes trigger the phosphorylation of RET receptors in neurons and initiate intracellular signaling pathways, which suppress mitochondrial fission and reduce oxidative stress. These changes inhibit caspase regulated cell apoptosis and promote neuronal survival and brain recovery after ischemic stroke

Article Snippet: The primary antibodies include a mouse anti-GFAP monoclonal antibody (1:300; Cat. No. MAB360, Sigma), a rabbit anti-GDNF polyclonal antibody (1:300, Cat. No. SC-328, Santa Cruz Biotechnology), a rabbit anti-Iba 1 polyclonal antibody (1:300, Cat. No. 019-19741, Fujifilm).

Techniques: Membrane, Phospho-proteomics, Protein-Protein interactions