rabbit polyclonal anti-gdnf antibody  (Thermo Fisher)

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

Journal: PeerJ

Article Title: Stress-induced changes in cognitive function and intestinal barrier integrity can be ameliorated by venlafaxine and synbiotic supplementations

doi: 10.7717/peerj.17033

Figure Lengend Snippet: Histological images of hippocampus (A–E) and pyknotic index in the dentate gyrus of the hippocampus in male rats under stress exposures (F). The black arrow indicates a normal granular cell, characterized by a dark-staining nucleus and scanty cytoplasm. The black asterisk indicates pyknotic nuclei, small condensed, dark-staining nuclei with eosinophilic cytoplasm. Relative glial cell-derived neurotrophic factor (GDNF) protein in hippocampus in stressed rats (G) and the representative expression of GDNF to GAPDH (H). * p < 0.05, ** p < 0.01 and *** p < 0.001 compared to vehicle-treated control rats. † p < 0.05 compared to vehicle-treated stressed rats ( n = 4 rats/group). Con, control; Str, stressed+vehicle; Str+Vlx, stressed+venlafaxine; Str+Syn, stressed+synbiotic supplement; Str+Vlx+Syn, stressed+venlafaxine+synbiotic supplement.

Article Snippet: These membranes were then subjected to overnight incubation at 4 °C with either a 1:1,000 dilution of rabbit polyclonal anti-GDNF antibody (catalog no. Cat #PA5-89957; Invitrogen, Thermo Fisher Scientific, Waltham, MA, USA), a 1:5,000 dilution of rabbit polyclonal anti-β-actin antibody (catalog no. AB8227; Abcam, Cambridge, UK) or GAPDH antibody (catalog no. Cat #PA1-988; Invitrogen, Thermo Fisher Scientific, Waltham, MA, USA).

Techniques: Staining, Derivative Assay, Expressing, Control

Journal: PeerJ

Article Title: Stress-induced changes in cognitive function and intestinal barrier integrity can be ameliorated by venlafaxine and synbiotic supplementations

doi: 10.7717/peerj.17033

Figure Lengend Snippet: Histological images of ileum (A–E) and inflammatory scores in male rats under stress exposures (F). The morphology of the ileum revealed desquamation of the villi (black star), villous edema (black triangle), loss of villi (asterisks) and layer separation (black arrow). 70-kDa FITC-dextran intestinal permeability (G), relative glial cell-derived neurotrophic factor (GDNF) protein in ileum in stressed rats (H) and the representative expression of GDNF beta actin (I). * p < 0.05, ** p < 0.01 and *** p < 0.001 compared to vehicle-treated control rats. † p < 0.05 and ††† p < 0.001 compared to vehicle-treated stressed rats ( n = 4 rats/group). Con, control; Str, stressed+vehicle; Str+Vlx, stressed+venlafaxine; Str+Syn, stressed+synbiotic supplement; Str+Vlx+Syn, stressed+venlafaxine+synbiotic supplement.

Article Snippet: These membranes were then subjected to overnight incubation at 4 °C with either a 1:1,000 dilution of rabbit polyclonal anti-GDNF antibody (catalog no. Cat #PA5-89957; Invitrogen, Thermo Fisher Scientific, Waltham, MA, USA), a 1:5,000 dilution of rabbit polyclonal anti-β-actin antibody (catalog no. AB8227; Abcam, Cambridge, UK) or GAPDH antibody (catalog no. Cat #PA1-988; Invitrogen, Thermo Fisher Scientific, Waltham, MA, USA).

Techniques: Permeability, Derivative Assay, Expressing, Control

Journal: Brain Sciences

Article Title: Overexpression of GDNF in Spinal Cord Attenuates Morphine Analgesic Tolerance in Rats with Bone Cancer Pain

doi: 10.3390/brainsci12091188

Figure Lengend Snippet: Chronic morphine reduced the level of GDNF in the spinal cord of BCP rats. ( A , B ) The immunochemistry test demonstrated that GDNF was located in the superficial laminae of the spinal cord. Compared with the sham operated rats treated with normal saline (sham+NS), the expression of GDNF was reduced in the BCP rats treated with normal saline (BCP+NS) and morphine (BCP+MT). n = 3 rats. *** p < 0.001, significantly different from sham +NS group. # p < 0.05, significantly different from the BCP+NS group. Scale bar = 200 μm. ( C ) Western blot demonstrated that expression of GDNF was decreased in BCP+MT rats compared with both the sham+NS and BCP+NS rats. n = 4 rats. *** p < 0.001, significantly different from the sham+NS group. # p < 0.05, significantly different from the BCP+NS group. ( D ) Western blot demonstrated that expression of GDNF was time-dependently reduced after chronic morphine treatment. n = 4 rats. ** p < 0.01, significantly different from the expression on Day 1.

Article Snippet: The sections of spinal cord were blocked with 10% normal donkey serum and 0.1% Triton X-100 in PBS for 1 h at 24 °C and then treated with rabbit anti-GDNF polyclonal antibody (1:100, Santa Cruz), mouse anti-MOR antibody (1:100, Abcam), and incubated at 4 °C in the refrigerator overnight.

Techniques: Saline, Expressing, Western Blot

Journal: Brain Sciences

Article Title: Overexpression of GDNF in Spinal Cord Attenuates Morphine Analgesic Tolerance in Rats with Bone Cancer Pain

doi: 10.3390/brainsci12091188

Figure Lengend Snippet: LV-GDNF intrathecal injection upregulated GDNF in the spinal cord of MT rats with BCP. ( A ) GFP expression was detected in the spinal cord of bone cancer pain and morphine tolerance rats treated with lentivirus-mediated GDNF (BM+LV-GDNF) and lentivirus of negative control (BM+LV-NC), but not in the BM rats treated with normal saline (BM+NS). Scale bar = 200 μm. ( B,C ) The overexpression of GDNF was detected by the immunochemistry test and Western blot in the spinal cord of the BM+LV-GDNF rats. n = 4 rats. *** p < 0.001, significantly different from the BM+LV-GDNF group.

Article Snippet: The sections of spinal cord were blocked with 10% normal donkey serum and 0.1% Triton X-100 in PBS for 1 h at 24 °C and then treated with rabbit anti-GDNF polyclonal antibody (1:100, Santa Cruz), mouse anti-MOR antibody (1:100, Abcam), and incubated at 4 °C in the refrigerator overnight.

Techniques: Injection, Expressing, Negative Control, Saline, Over Expression, Western Blot

Journal: Brain Sciences

Article Title: Overexpression of GDNF in Spinal Cord Attenuates Morphine Analgesic Tolerance in Rats with Bone Cancer Pain

doi: 10.3390/brainsci12091188

Figure Lengend Snippet: Upregulation of GDNF alleviated morphine tolerance in bone cancer pain rats. The mechanical ( A ) and thermal ( B ) withdrawal threshold indicated that LV-GDNF injection alleviated the morphine analgesic tolerance in BCP rats. n = 8 rats. * p < 0.05, ** p < 0.01, *** p < 0.001, significantly different from bone cancer pain and morphine tolerance rats treated with normal saline (BM+NS). ( C ) The percentage of the maximum possible effect (%MPE) of morphine was detected by the tail-flick latency test in the bone cancer pain rats. n = 8 rats. ** p < 0.01, *** p < 0.001, significantly different from BM rats treated with normal saline (BM+NS). ( D ) Time course of morphine analgesic effect in BM rats was detected by the thermal paw withdrawal latency on POD 15. n = 8 rats. ** p < 0.01, *** p < 0.001, significantly different from BM+NS group. ###, p < 0.001, compared with the corresponding baseline.

Article Snippet: The sections of spinal cord were blocked with 10% normal donkey serum and 0.1% Triton X-100 in PBS for 1 h at 24 °C and then treated with rabbit anti-GDNF polyclonal antibody (1:100, Santa Cruz), mouse anti-MOR antibody (1:100, Abcam), and incubated at 4 °C in the refrigerator overnight.

Techniques: Injection, Saline, Tail Flick Test

Journal: Brain Sciences

Article Title: Overexpression of GDNF in Spinal Cord Attenuates Morphine Analgesic Tolerance in Rats with Bone Cancer Pain

doi: 10.3390/brainsci12091188

Figure Lengend Snippet: Upregulation of GDNF recovered MOR expression in morphine tolerance rats with bone cancer pain. ( A ) MOR expression was reduced in the spinal cord of bone cancer pain rats treated with normal saline (BCP+NS), and further downregulated in BCP rats with morphine tolerance (BCP+MT). Scale bar = 200 μm. ( B ) The double immunochemistry test demonstrated that GDNF and MOR were coexpressed in the spinal cord of rats. Scale bar = 200 μm. ( C , D ) The expression of MOR in the spinal cord of morphine tolerance rats with bone cancer pain was restored by LV-GDNF injection, but not LV-NC intrathecal injection. n = 4 rats. ** p < 0.01, significantly different from the LV-NC group.

Article Snippet: The sections of spinal cord were blocked with 10% normal donkey serum and 0.1% Triton X-100 in PBS for 1 h at 24 °C and then treated with rabbit anti-GDNF polyclonal antibody (1:100, Santa Cruz), mouse anti-MOR antibody (1:100, Abcam), and incubated at 4 °C in the refrigerator overnight.

Techniques: Expressing, Saline, Injection

Journal: Glia

Article Title: GLAST-CreER T2 mediated deletion of GDNF increases brain damage and exacerbates long-term stroke outcomes after focal ischemic stroke in mouse model.

doi: 10.1002/glia.23848

Figure Lengend Snippet: FIGURE 1 GDNF is dramatically upregulated after PT. (a) WB images of GDNF at different times after PT. (b) WB analysis of GDNF showing dramatic increase after PT. Data were analyzed as fold change of GDNF/β-actin compared with normal control mice. N = 3–5 mice for each time point. *, #, p < .05, **, ##, p < .005 compared with the control group, ANOVA test. (c) Confocal images of immunostaining show the colocalization of GDNF with GFAP+ RAs (*) in the PIR 4 days after PT (2015). The bottom panels are the high-resolution images of the boxed region A1 and the orthogonal analysis. C, contralateral hemisphere; I, ipsilateral hemisphere; IC, ischemic core; PIR, peri-infarct region

Article Snippet: The primary antibodies include a rabbit anti-GDNF polyclonal antibody (1:200; SC-328, Santa Cruz Biotechnology, Santa Cruz, CA), a rabbit anti-Ki67 (1:600, AB9260, Sigma), a rabbit anti-doublecortin (DCX) (1:1,000; Ab18723, Abcam, MA.

Techniques: Control, Immunostaining

Journal: Glia

Article Title: GLAST-CreER T2 mediated deletion of GDNF increases brain damage and exacerbates long-term stroke outcomes after focal ischemic stroke in mouse model.

doi: 10.1002/glia.23848

Figure Lengend Snippet: FIGURE 2 Generation and characterization of GLAST-GDNF−/−cKO mice. (a) Breeding scheme to obtain tamoxifen (TAM) inducible and astrocyte-specific GLAST-GDNF−/−cKO mice. (b) Gel images of genotyping. (c) Experimental timeline. (d) Average body weights of total, male and female of GDNFf/f and GLAST-GDNF−/−cKO mice. (e) Nissl staining of brain sections showing cortical and hippocampal structure. (f,g) Photographs of whole brain (f) and average brain weight (g). (h,i) WB analysis of GDNF levels in the cortex and hippocampus of GDNFf/f and GLAST-GDNF−/−cKO mice using mouse anti-GDNF antibody. Summary data were obtained from the averaged values of N = 5 mice for each group. J-K) WB analysis of pre- and post-synaptic proteins in the cortex (and hippocampus). Summary data were obtained from the averaged values of N = 5 mice for each group

Article Snippet: The primary antibodies include a rabbit anti-GDNF polyclonal antibody (1:200; SC-328, Santa Cruz Biotechnology, Santa Cruz, CA), a rabbit anti-Ki67 (1:600, AB9260, Sigma), a rabbit anti-doublecortin (DCX) (1:1,000; Ab18723, Abcam, MA.

Techniques: Staining

Journal: Glia

Article Title: GLAST-CreER T2 mediated deletion of GDNF increases brain damage and exacerbates long-term stroke outcomes after focal ischemic stroke in mouse model.

doi: 10.1002/glia.23848

Figure Lengend Snippet: FIGURE 3 Deletion of astrocytic GDNF reduces adult neurogenesis in DG under normal conditions. (a) Experimental protocol for Brdu injection. (b) Immunostaining of Brdu and NeuN for the illustration of adult neurogenesis in the dentate gyrus (DG) of hippocampus. SGZ- subgranular zone; GCL-granular cell layer; ML-molecular layer. (c,e,g) Fluorescent images of Brdu/Dapi (c), Ki67/Dapi (e) and DCX/Dapi double staining in DG of GDNFf/f and GLAST-GDNF−/−cKO mice. (d,f,h) Summary data of Brdu+ cells (d), Ki67+ cells (f) and DCX+ neuroblasts (h). Four- weeks old GLAST-CreERT2:GDNFf/f mice were injected with TAM (100 mg/kg) and sacrificed 2 weeks after last injection (also see Figure 2). Data were averaged from N = 4 mice for each genotype and 2 slices of each mouse. *p < .05, t test

Article Snippet: The primary antibodies include a rabbit anti-GDNF polyclonal antibody (1:200; SC-328, Santa Cruz Biotechnology, Santa Cruz, CA), a rabbit anti-Ki67 (1:600, AB9260, Sigma), a rabbit anti-doublecortin (DCX) (1:1,000; Ab18723, Abcam, MA.

Techniques: Injection, Immunostaining, Double Staining

Journal: Glia

Article Title: GLAST-CreER T2 mediated deletion of GDNF increases brain damage and exacerbates long-term stroke outcomes after focal ischemic stroke in mouse model.

doi: 10.1002/glia.23848

Figure Lengend Snippet: FIGURE 4 GLAST-mediated GDNF deletion reduces neuronal stem cell and progenitor cell survival in DG under normal conditions. (a) Brdu injection and experimental protocol. (b–e) Summary results of immunostaining for Brdu+ cells in the SGZ (b) and GCL (c), Brdu+Sox2+ cells (d) and Brdu+NeurD1+ cells (e) in SGZ 4 weeks after Brdu injection. Data in (b–e) were averaged from N = 6 mice for each genotype and 2 slices for each mouse. *p < .05, t test. (f–i) Confocal images of Brdu+ and NeuroD1 (f,g), and Brdu and Sox2 double staining (h,i) of GDNFf/f mice (f) and GLAST- GDNF−/−cKO mice (g) 28 days after TAM administration. Images in the right panels are the high-resolution images of the boxed regions in the left panels. Arrowheads point to the cells with colocalization of indicated markers

Article Snippet: The primary antibodies include a rabbit anti-GDNF polyclonal antibody (1:200; SC-328, Santa Cruz Biotechnology, Santa Cruz, CA), a rabbit anti-Ki67 (1:600, AB9260, Sigma), a rabbit anti-doublecortin (DCX) (1:1,000; Ab18723, Abcam, MA.

Techniques: Injection, Immunostaining, Double Staining

Journal: Glia

Article Title: GLAST-CreER T2 mediated deletion of GDNF increases brain damage and exacerbates long-term stroke outcomes after focal ischemic stroke in mouse model.

doi: 10.1002/glia.23848

Figure Lengend Snippet: FIGURE 5 Deletion of GDNF in astrocyte increases ischemic infarction and neuronal degeneration after PT. (a) Representative Nissl staining of brain sections showing brain infarct areas of GDNFf/f and GLAST-GDNF−/−cKO mice 2 days after PT. The dashed lines outline the infarct regions. IC-ischemic core. (b,c) Infarct areas and infarct volumes at different days after PT. Data were averaged from N = 6 mice for each group 2 days after PT, N = 6 GNDFf/f mice and 7 GLAST-GDNF−/−mice for 4 days after PT, and N = 6 mice for each group 14 days after PT. White dash lines outline the total damage area, and the area between red and white dash lines is the area of damaged hipppocampus. (d) The volume percentage of hippocampal damage. Data were averaged from N = 6 mice for each group 2 days after PT, N = 6 GNDFf/f mice and 7 GLAST- GDNF−/−mice for 4 days after PT. (e,f) Representative images of FJB staining (d) and the densities of FJB+ cells (e) in the PIR 2 days after PT. N = 6 mice for each group and 2 slice for each mouse. (g,h) Confocal images of double immunostaining of GFAP and GDNF in control and GDNF cKO mice 4 days after PT. Arrows indicate colocalization of GDNF and GFAP in reactive astrocytes while arrow heads reactive astrocytes without expressing GDNF. (i,j) WB analysis of GDNF in the cortex 24 days after PT. N = 7 mice for each group. *p < .05, **p < .005, ***p < .0005, t test

Article Snippet: The primary antibodies include a rabbit anti-GDNF polyclonal antibody (1:200; SC-328, Santa Cruz Biotechnology, Santa Cruz, CA), a rabbit anti-Ki67 (1:600, AB9260, Sigma), a rabbit anti-doublecortin (DCX) (1:1,000; Ab18723, Abcam, MA.

Techniques: Staining, Double Immunostaining, Control, Expressing

Journal: Glia

Article Title: GLAST-CreER T2 mediated deletion of GDNF increases brain damage and exacerbates long-term stroke outcomes after focal ischemic stroke in mouse model.

doi: 10.1002/glia.23848

Figure Lengend Snippet: FIGURE 6 Deletion of astrocytic GDNF attenuates cell proliferation in the PIR after PT. (a–d) Fluorescent images of Brdu+ (a,b) and Ki67+ (c,d) cells in the PIR in GDNFf/f and GLAST-GDNF−/−cKO mice 2 days after PT. (b) and (d) are the high resolution images of the boxed regions in (a) and (b). PIR-Peri infarct region, IC-ischemic core. (e–g) Summary of the densities of Brdu+ and Ki67+ cells in the PIR 2 (e,f) and 4 (g,h) days after PT. Mice were injected with Brdu immediately after PT and sacrificed 2 and 4 days later. N = 3 mice and 2 slices for each mouse in each group. *p < .05, t test

Article Snippet: The primary antibodies include a rabbit anti-GDNF polyclonal antibody (1:200; SC-328, Santa Cruz Biotechnology, Santa Cruz, CA), a rabbit anti-Ki67 (1:600, AB9260, Sigma), a rabbit anti-doublecortin (DCX) (1:1,000; Ab18723, Abcam, MA.

Techniques: Injection

Journal: Glia

Article Title: GLAST-CreER T2 mediated deletion of GDNF increases brain damage and exacerbates long-term stroke outcomes after focal ischemic stroke in mouse model.

doi: 10.1002/glia.23848

Figure Lengend Snippet: FIGURE 7 Deletion of astrocytic GDNF attenuates reactive gliosis in the PIR after PT. (a–c) Double immunostaining images of Brdu and GFAP in the PIR of GDNFf/f and GLAST-GDNF−/−cKO mice at 2 (a), 4 (b), and 14 (c) days after PT. Scale bar is 50 μm in (a–c). (d) Quantification of GFAP immunofluorescent signal 2 days after PT. (e) Densities of GFAP+ RAs. (f) Brdu+ cells. (g) Brdu+GFAP+ cells. (h) The ratio of Brdu+GFAP+ cells to GFAP+ cells. (i) the ratio of Brdu+GFAP+ cells to Brdu+ cells. Data in (d–i) were averaged values from 3 to 4 mice and 2 slices for each mouse. *p < .05, **p < .005, t test

Article Snippet: The primary antibodies include a rabbit anti-GDNF polyclonal antibody (1:200; SC-328, Santa Cruz Biotechnology, Santa Cruz, CA), a rabbit anti-Ki67 (1:600, AB9260, Sigma), a rabbit anti-doublecortin (DCX) (1:1,000; Ab18723, Abcam, MA.

Techniques: Double Immunostaining

Journal: Glia

Article Title: GLAST-CreER T2 mediated deletion of GDNF increases brain damage and exacerbates long-term stroke outcomes after focal ischemic stroke in mouse model.

doi: 10.1002/glia.23848

Figure Lengend Snippet: FIGURE 8 Deletion of astrocytic GDNF increase oxidative stress. (a,b) Double immunostaining of G6PD and GFAP in the PIR (upper panel) and contralateral side (lower panel) of GDNFf/f mice (a) and GLAST-GDNF−/−mice (b) 4 days after PT. Notice that G6PD is highly colocalized with GFAP in PIR in the control mice and the reduction of G6PD expression in the cKO mice. (c) Summary data of G6PD+ cells (left), G6PD +GFAP+ RAs (middle) and the ratio of G6PD+RAs to total GFAP+ RAs, that is, G6PD+GFAP+/GFAP+ (right) in GDNFf/f mice and GLAST- GDNF−/−mice 4 days after PT. The data were averaged from four slices of each genotype. (d,e) Fluorescent images of DHE and Dapi in the PIR (upper panel) of GDNFf/f mice (d) and GLAST-GDNF−/−mice (e) 2 days after PT. The bottom panels are the high-resolution images of the boxed region of upper panels. Notice the increases in DHE signal in the cKO mice. (f) Summary data of DHE fluorescent intensity in the PIR of GDNFf/f

Article Snippet: The primary antibodies include a rabbit anti-GDNF polyclonal antibody (1:200; SC-328, Santa Cruz Biotechnology, Santa Cruz, CA), a rabbit anti-Ki67 (1:600, AB9260, Sigma), a rabbit anti-doublecortin (DCX) (1:1,000; Ab18723, Abcam, MA.

Techniques: Double Immunostaining, Control, Expressing

Journal: Glia

Article Title: GLAST-CreER T2 mediated deletion of GDNF increases brain damage and exacerbates long-term stroke outcomes after focal ischemic stroke in mouse model.

doi: 10.1002/glia.23848

Figure Lengend Snippet: FIGURE 9 Deletion of astrocytic GDNF increases behavioral deficits after PT. (a–d) Four motor behavioral tests, that is, hanging wire (a), cylinder (b), rotorad (c), and grip force (d) tests were conducted on GDNFf/f and GLAST-GDNF−/−cKO mice before and different times after PT. (e–k) Open field test for the evaluation of the general motor activity and anxiety. Time courses of six parameters from 10 min open field tests including total travel distance (e), corner distance (f), total immobile time (g), the ratio of corner travel distance to total travel distance (h), the time to explore the corners (i), the ratio of center distance to total distance (j), and the time to explore the center (k). The value of Day 0 in each test is the data before PT for ischemic mouse group. Mouse numbers were indicated in the figures. *p < .05, **p < .005, ***p < .0005, ANOVA test [Color figure can be viewed at wileyonlinelibrary.com]

Article Snippet: The primary antibodies include a rabbit anti-GDNF polyclonal antibody (1:200; SC-328, Santa Cruz Biotechnology, Santa Cruz, CA), a rabbit anti-Ki67 (1:600, AB9260, Sigma), a rabbit anti-doublecortin (DCX) (1:1,000; Ab18723, Abcam, MA.

Techniques: Activity Assay