human gfra1 r d systems  (R&D Systems)

Journal: Frontiers in Pharmacology

Article Title: Enhanced therapeutic potential of paeoniflorin and vitamin B12 in intracerebropeduncle ethidium bromide-induced multiple sclerosis-like pathology

doi: 10.3389/fphar.2026.1792674

Figure Lengend Snippet: The figures illustrate the docking interaction of paeoniflorin with the GDNF receptor (PDB ID: 1AGQ). (A) Shows a surface representation of the GDNF receptor, with the active site highlighted in yellow, where the ligand paeoniflorin (red) is bound. (B) 2D interaction diagram displays key interactions between paeoniflorin and the receptor, with hydrogen bonds and hydrophobic interactions clearly indicated. (C) Provides a 3D view of the docking, showing paeoniflorin (blue) interacting with GDNF, with hydrogen bonds represented by green dashed lines and key regions labeled. (D) Offers a zoomed-in view of the binding site, emphasizing hydrogen bonds and hydrophobic contacts, depicted within a ribbon structure for clarity.

Article Snippet: The ELISA kits for evaluating cellular and molecular targets included GDNF [E-EL-H1495; Elabscience GFRA1 [PKSH033670; Elabscience]; RET [AN00810P; Elabscience], AKT [E-EL-R0807 98T, Elabscience, Wuhan, China] ( ); ERK1/2 [E-AB-70292; Elabscience] ( ); and GSK3-Beta [KLR0989, KRISHGEN, Maharashtra, India] ( ).

Techniques: Labeling, Binding Assay

Journal: Frontiers in Pharmacology

Article Title: Enhanced therapeutic potential of paeoniflorin and vitamin B12 in intracerebropeduncle ethidium bromide-induced multiple sclerosis-like pathology

doi: 10.3389/fphar.2026.1792674

Figure Lengend Snippet: (A–H) PNN neuroprotective role in mitigating EBRO-induced alterations in levels of cellular and molecular targets in MS rat model: GDNF (A) , GFRA1 (B) , AKT (C) , ERK1/2 (D) , GSK3-Beta (E) , in brain homogenates, and GDNF, GFRA1, AKT, ERK1/2, GSK3-Beta in CSF levels (F–H) . Statistical analysis was performed using a one-way ANOVA followed by Tukey’s post hoc test to determine significant differences among groups (A–H) . Data were presented as mean ± standard deviation (SD), with statistical significance set at p < 0.01. Each experimental group consisted of eight wistar rats (n = 8). β v/s Sham Control, Vehicle Control, and PNN Perse; δ v/s EBRO; δα1 v/s EBRO + PNN50; δα2 v/s EBRO + PNN100, EBRO + PNN50; and δα3 v/s EBRO + VB12 (30), EBRO + PNN100, EBRO + PNN50. To identify significant differences between groups, a one-way ANOVA and Tukey’s post hoc test were used for statistical analysis (A–H) . The statistical significance level was set at p < 0.01, and the data were displayed as mean ± standard deviation (SD). There were eight wistar rats (n = 8) in each experimental group. β v/s Sham Control, Vehicle Control, and PNN Perse; δ v/s EBRO; δα1 v/s EBRO + PNN50; δα2 v/s EBRO + PNN100, EBRO + PNN50; and δα3 v/s EBRO + VB12 (30), EBRO + PNN100, EBRO + PNN50.

Article Snippet: The ELISA kits for evaluating cellular and molecular targets included GDNF [E-EL-H1495; Elabscience GFRA1 [PKSH033670; Elabscience]; RET [AN00810P; Elabscience], AKT [E-EL-R0807 98T, Elabscience, Wuhan, China] ( ); ERK1/2 [E-AB-70292; Elabscience] ( ); and GSK3-Beta [KLR0989, KRISHGEN, Maharashtra, India] ( ).

Techniques: Standard Deviation, Control

Journal: Frontiers in Pharmacology

Article Title: Enhanced therapeutic potential of paeoniflorin and vitamin B12 in intracerebropeduncle ethidium bromide-induced multiple sclerosis-like pathology

doi: 10.3389/fphar.2026.1792674

Figure Lengend Snippet: (A–H) PNN neuroprotective role in mitigating EBRO-induced alterations in levels of cellular and molecular targets in MS rat model: GDNF (A) , GFRA1 (B) , AKT (C) , ERK1/2 (D) , GSK3-Beta (E) , in brain homogenates, and GDNF, GFRA1, AKT, ERK1/2, GSK3-Beta in CSF levels (F–H) . Statistical analysis was performed using a one-way ANOVA followed by Tukey’s post hoc test to determine significant differences among groups (A–H) . Data were presented as mean ± standard deviation (SD), with statistical significance set at p < 0.01. Each experimental group consisted of eight wistar rats (n = 8). β v/s Sham Control, Vehicle Control, and PNN Perse; δ v/s EBRO; δα1 v/s EBRO + PNN50; δα2 v/s EBRO + PNN100, EBRO + PNN50; and δα3 v/s EBRO + VB12 (30), EBRO + PNN100, EBRO + PNN50. To identify significant differences between groups, a one-way ANOVA and Tukey’s post hoc test were used for statistical analysis (A–H) . The statistical significance level was set at p < 0.01, and the data were displayed as mean ± standard deviation (SD). There were eight wistar rats (n = 8) in each experimental group. β v/s Sham Control, Vehicle Control, and PNN Perse; δ v/s EBRO; δα1 v/s EBRO + PNN50; δα2 v/s EBRO + PNN100, EBRO + PNN50; and δα3 v/s EBRO + VB12 (30), EBRO + PNN100, EBRO + PNN50.

Article Snippet: The ELISA kits for evaluating cellular and molecular targets included GDNF [E-EL-H1495; Elabscience GFRA1 [PKSH033670; Elabscience]; RET [AN00810P; Elabscience], AKT [E-EL-R0807 98T, Elabscience, Wuhan, China] ( ); ERK1/2 [E-AB-70292; Elabscience] ( ); and GSK3-Beta [KLR0989, KRISHGEN, Maharashtra, India] ( ).

Techniques: Standard Deviation, Control

Journal: Andrology

Article Title: Effects of neonatal hypothyroidism on testicular development and undifferentiated spermatogonia in prepubertal rats

doi: 10.1111/andr.70116

Figure Lengend Snippet: Hematoxylin and eosin staining and immunostaining images of testes with anti‐DDX4, anti‐GFRA1, and anti‐SOX9 antibodies in the control, 0.001% 6‐n‐propyl‐2‐thiouracil (PTU), 0.01% PTU, and 0.03% PTU groups on postnatal day 20 in transient hypothyroidism (tHT) model rats. In all groups, the development of the seminiferous tubule lumen structure was observed; however, as the PTU concentration increased, the proliferation and differentiation of spermatocytes into mature stages of the cell life cycle were not observed, and the seminiferous tubules were narrower, with wider surrounding interstitial areas (A‒D). Immunostaining with the anti‐DDX4 antibody (E‒H) revealed positive germ cells with large, round, or semicircular nuclei located in the marginal region adjacent to the basement membrane of the seminiferous tubules. The number of positive cells per cross‐section of the seminiferous tubules was higher in the three PTU‐treated groups than in the control group. Immunostaining with the anti‐GFRA1 antibody (I‒L) showed positive cells in the marginal region of the seminiferous tubules, with a higher number of positive cells per cross‐section in the three PTU‐treated groups. Staining with the anti‐SOX9 antibody (M‒P) revealed positive Sertoli cells with spindle‐shaped nuclei at the periphery of the seminiferous tubules, displaying similar staining patterns in all groups. Negative control for DDX4, GFRA1, and SOX9 immunostaining in the testis of the tHT model on postnatal day 20 (Q‒X). Scale bar = 100 µm.

Article Snippet: Paraffin‐embedded testicular sections (5 μm thick) were deparaffinized, rehydrated, and subjected to antigen retrieval using 10 mM sodium citrate buffer (pH 6.0) at 95°C for 40 min. After cooling to room temperature (20°C–25°C) and blocking with 5% skim milk diluted in phosphate‐buffered saline with Tween 20 for 1 h, the sections were incubated overnight at 4°C with primary antibodies against GFRA1 (1:100; AF560; R&D Systems) and Ki‐67 (1:1000; ab15580; Abcam).

Techniques: Staining, Immunostaining, Control, Concentration Assay, Membrane, Negative Control

Journal: Andrology

Article Title: Effects of neonatal hypothyroidism on testicular development and undifferentiated spermatogonia in prepubertal rats

doi: 10.1111/andr.70116

Figure Lengend Snippet: Hematoxylin and eosin staining and images of immunostaining images of testes with anti‐DDX4, anti‐GFRA1, and anti‐SOX9 antibodies in the control group and 0.001% 6‐n‐propyl‐2‐thiouracil (PTU), 0.01% PTU, and 0.03% PTU groups on postnatal day 7 in both the continuous hypothyroidism (cHT) and transient hypothyroidism (tHT) models. Within the seminiferous tubules, germ cells and Sertoli cells were similarly observed across all groups; however, as the PTU concentration increased, the seminiferous tubules appeared narrower, and the surrounding interstitial areas appeared wider (A‒D). Immunostaining with the anti‐DDX4 (E‒H) and anti‐GFRA1 (I‒L) antibodies showed positive staining in germ cells, which exhibited large, round nuclei located within or at the periphery of the seminiferous tubules. Compared to the control group, the three PTU‐treated groups had a higher number of positive cells per cross‐section of the seminiferous tubules. Staining with the anti‐SOX9 antibody (M‒P) revealed positive Sertoli cells with spindle‐shaped nuclei at the periphery of the seminiferous tubules, showing a similar staining pattern across all groups. Scale bar = 100 µm.

Article Snippet: Paraffin‐embedded testicular sections (5 μm thick) were deparaffinized, rehydrated, and subjected to antigen retrieval using 10 mM sodium citrate buffer (pH 6.0) at 95°C for 40 min. After cooling to room temperature (20°C–25°C) and blocking with 5% skim milk diluted in phosphate‐buffered saline with Tween 20 for 1 h, the sections were incubated overnight at 4°C with primary antibodies against GFRA1 (1:100; AF560; R&D Systems) and Ki‐67 (1:1000; ab15580; Abcam).

Techniques: Staining, Immunostaining, Control, Concentration Assay

Journal: Andrology

Article Title: Effects of neonatal hypothyroidism on testicular development and undifferentiated spermatogonia in prepubertal rats

doi: 10.1111/andr.70116

Figure Lengend Snippet: Hematoxylin and eosin staining and immunostaining images of testes with anti‐DDX4, anti‐GFRA1, and anti‐SOX9 antibodies in the control, 0.001% 6‐n‐propyl‐2‐thiouracil (PTU), 0.01% PTU, and 0.03% PTU groups on postnatal day 20 in continuous hypothyroidism (cHT) model rats. In the control group, the seminiferous tubules exhibited clear lumen formation, whereas no lumen formation was observed in any of the PTU‐treated groups (A‒D). These groups also showed markedly reduced seminiferous tubule diameters and expanded interstitial areas compared to the control group. Immunostaining with the anti‐DDX4 antibody (E‒H) revealed positive germ cells in all groups, but the number of DDX4‐positive cells per cross‐section was consistently lower in the PTU‐treated groups. Although GFRA1‐positive (I‒L) and SOX9‐positive (M‒P) cells were present in the PTU‐treated groups, the significant morphological differences in seminiferous tubules and developmental stages made direct quantitative comparisons with the control group inappropriate. Scale bar = 100 µm.

Article Snippet: Paraffin‐embedded testicular sections (5 μm thick) were deparaffinized, rehydrated, and subjected to antigen retrieval using 10 mM sodium citrate buffer (pH 6.0) at 95°C for 40 min. After cooling to room temperature (20°C–25°C) and blocking with 5% skim milk diluted in phosphate‐buffered saline with Tween 20 for 1 h, the sections were incubated overnight at 4°C with primary antibodies against GFRA1 (1:100; AF560; R&D Systems) and Ki‐67 (1:1000; ab15580; Abcam).

Techniques: Staining, Immunostaining, Control

Journal: Andrology

Article Title: Effects of neonatal hypothyroidism on testicular development and undifferentiated spermatogonia in prepubertal rats

doi: 10.1111/andr.70116

Figure Lengend Snippet: Comparison of positive cell counts for immunostaining (DDX4, GFRA1, and SOX9) among the four groups on postnatal days 7 and 20 in the transient hypothyroidism (tHT) model. On postnatal day 7, the three 6‐n‐propyl‐2‐thiouracil (PTU)‐treated groups exhibited a comparable increase in the number of DDX4‐positive cells per cross‐section of the seminiferous tubules compared to the control group (A and C). However, no significant differences were observed among the four groups for SOX9 (E). Furthermore, on postnatal day 20, the three PTU‐treated groups showed a significant increase in the average number of DDX4‐positive cells in the peripheral region per cross‐section of the seminiferous tubules compared to the control group (B). Additionally, the 0.01% PTU and 0.03% PTU groups exhibited a significant increase in the number of GFRA1‐positive cells per cross‐section compared to that in the control group (D). As with postnatal day 7, no significant differences were observed among the four groups for SOX9 (F). An asterisk indicates a significant difference from the control group ( * p < 0.05).

Article Snippet: Paraffin‐embedded testicular sections (5 μm thick) were deparaffinized, rehydrated, and subjected to antigen retrieval using 10 mM sodium citrate buffer (pH 6.0) at 95°C for 40 min. After cooling to room temperature (20°C–25°C) and blocking with 5% skim milk diluted in phosphate‐buffered saline with Tween 20 for 1 h, the sections were incubated overnight at 4°C with primary antibodies against GFRA1 (1:100; AF560; R&D Systems) and Ki‐67 (1:1000; ab15580; Abcam).

Techniques: Comparison, Immunostaining, Control

Journal: Andrology

Article Title: Effects of neonatal hypothyroidism on testicular development and undifferentiated spermatogonia in prepubertal rats

doi: 10.1111/andr.70116

Figure Lengend Snippet: GFRA1/Ki‐67 double immunofluorescence staining in the transient hypothyroidism (tHT) model (0.01% 6‐n‐propyl‐2‐thiouracil [PTU]) on postnatal day 7. Two GFRA1‐positive cells (green) were observed along the periphery of the seminiferous tubule located at the center of the image. Ki‐67‐positive nucleoli (red) were present within the nuclei of these cells. Scale bar = 50 µm.

Article Snippet: Paraffin‐embedded testicular sections (5 μm thick) were deparaffinized, rehydrated, and subjected to antigen retrieval using 10 mM sodium citrate buffer (pH 6.0) at 95°C for 40 min. After cooling to room temperature (20°C–25°C) and blocking with 5% skim milk diluted in phosphate‐buffered saline with Tween 20 for 1 h, the sections were incubated overnight at 4°C with primary antibodies against GFRA1 (1:100; AF560; R&D Systems) and Ki‐67 (1:1000; ab15580; Abcam).

Techniques: Double Immunofluorescence Staining

Journal: PLOS One

Article Title: GDNF receptor GFRα1 is necessary for the maintenance of dopaminergic neurons in the adult substantia nigra

doi: 10.1371/journal.pone.0331369

Figure Lengend Snippet: (A-E’) Representative confocal images of the dSTR showing RNAscope mRNA signal for Gfra1 (red), immunolabeling for DARPP-32 (green), and counterstaining with DAPI (blue) in coronal brain sections from C57BL6/J mouse at P0 (A), P5 (B), P15(C), P30 (D) or P60 (E). Scale bar = 10 μm. (F) Expression levels (± SEM) of Gfra1 (N) mRNA quantified by RNAscope puncta analysis in the dSTR across postnatal developmental ages. Statistical comparisons among ages are described by the table beside. N = 8 dSTR from 4 mice for all ages except P0 (N = 10 dSTR from 5 mice). *p < 0,05; ***p < 0,001; One-way Anova. (G-G’) Representative confocal low magnification images of the striatum showing dTomato epifluorescence (red), immunolabeling for DARPP-32 (green) and counterstaining with DAPI (blue) in coronal brain sections of Gfr α 1 CreERT2/+ ; Rosa26 dTOM (GFRα1 +/- ) mouse injected with tamoxifen at 3 months. GFRα1 dTom+ cells can be observed predominantly in the lateral septum (LS) and sparsely in the ventral striatum (vSTR) but not in the dorsal striatum (dSTR). Scale bar = 400 μm.

Article Snippet: All animals were bred in a C57BL6/J background (Charles River, Netherlands) except from Gfra1 flox , which was kept in a CD1 background (Charles River, Netherlands).

Techniques: RNAscope, Immunolabeling, Expressing, Injection