glycine receptor  (Addgene inc)

Journal: Frontiers in Immunology

Article Title: Glycine receptors in circulating white blood cells regulated by neuroinflammation

doi: 10.3389/fimmu.2026.1749275

Figure Lengend Snippet: Identification and localization of GlyRα subunits in human PBMCs and CD14+ monocytes. (A) Relative mRNA expression of GlyRα1, α2, and α3 in CD14+ monocytes (white bars) and PBMCs (gray bars), normalized to β2-microglobulin housekeeping gene. GlyRα1 showed the highest expression, followed by GlyRα2 and α3. Data are presented as mean ± SEM (B) Quantification of PBMCs expressing 55.21 ± 2.8% of GlyRα1/2+ cells and 46.87 ± 3.4% of GlyRα3+ cells, with 43.12 ± 2.2% of GlyRα1/2+/GlyRα3+ PBMCs. GlyRα1/2+ cells were significantly more numerous than dual-positive cells (p=0.015*). Statistical significance was assessed using one-way ANOVA; bars depict mean ± SEM; *p<0.05. (C) Representative immunofluorescence images of PBMCs showing GlyRα1/2 (red), GlyRα3 (green), and nuclei stained with DAPI (blue, arrowhead). The merged panel shows the co-localization of GlyRα1/2 and GlyRα3, with a few cells negative for GlyRs.

Article Snippet: After blocking with 5% normal goat serum (NGS) for 1 hour, the slides and coverslips were stained with primary antibodies targeting human and mouse GlyRα1 (1:500, #NB300-113, Novus Bio, Littleton, CO), GlyRα3 (1:100, #75-417, Neuromab, UC Davis/NIH), CD11b (1:100, #Sc-1186, Santa Cruz Biotechnology), CD-68 (1:100, #Sc-20060, Santa Cruz Biotechnology), and Iba1 (1:400, #MABN92, Millipore Sigma) ( ) overnight at 4°C.

Techniques: Expressing, Immunofluorescence, Staining

Journal: Frontiers in Immunology

Article Title: Glycine receptors in circulating white blood cells regulated by neuroinflammation

doi: 10.3389/fimmu.2026.1749275

Figure Lengend Snippet: Identification of GlyRα subunits in mouse brain, spleen, bone marrow, and blood. Normalized mRNA expression levels of GlyRα1, α2, and α3 in mouse brain (A) , spleen (C) , bone marrow (BM; E ), and blood (G) show the highest expression of GlyRα2, followed by GlyRα3 and α1. Statistical significance was determined using one-way ANOVA followed by Tukey’s post hoc test. Data are shown as mean ± SEM; p<0.01** (n=4-6 mice per group). (B) Representative immunofluorescence images of brain sections stained for GlyRα1/2 ( red ), GlyRα3 ( green ), and nuclei (DAPI, blue ). GlyRα1/2 and α3 signals show co-expression within neurons of the hippocampus. (D) Spleen sections display strong GlyRα1/2 (red) staining in both red (arrow) and white pulps (arrow head), whereas GlyRα3 (green) staining is localized to the macrophage zone of red pulp. (F, H) Immunostaining of BM and WBCs shows strong membrane expression of GlyRα1/2 and α3. GlyRα3 staining marks distinct cell populations distributed in the cytoplasm and membrane. The merged image shows GlyRα1/2 and α3 co-localization, indicating heterogeneity in receptor subunit expression among different cell BM and WBC subsets.

Article Snippet: After blocking with 5% normal goat serum (NGS) for 1 hour, the slides and coverslips were stained with primary antibodies targeting human and mouse GlyRα1 (1:500, #NB300-113, Novus Bio, Littleton, CO), GlyRα3 (1:100, #75-417, Neuromab, UC Davis/NIH), CD11b (1:100, #Sc-1186, Santa Cruz Biotechnology), CD-68 (1:100, #Sc-20060, Santa Cruz Biotechnology), and Iba1 (1:400, #MABN92, Millipore Sigma) ( ) overnight at 4°C.

Techniques: Expressing, Immunofluorescence, Staining, Immunostaining, Membrane

Journal: Frontiers in Immunology

Article Title: Glycine receptors in circulating white blood cells regulated by neuroinflammation

doi: 10.3389/fimmu.2026.1749275

Figure Lengend Snippet: Blood-based GlyR response under different inflammatory conditions. (A) Representative immunofluorescence images showing GlyRα1/2 (red), GlyRα3 (green), and DAPI (blue) staining in WBCs from control, systemic- and neuroinflammation groups. Scale bar: 10µm (B) . Fold-change expression of GlyRα1, α2, and α3 in circulating WBCs (n=4-6 mice per group) showed that neuroinflammation significantly upregulated all three GlyRα subunits, while systemic inflammation selectively increased GlyRα1 and α2 expression. This suggests enhanced peripheral glycinergic activation in response to neuroinflammation. The experiment was repeated twice in duplicates. Statistical significance was assessed by one-way ANOVA followed by Tukey’s post hoc test (*p<0.05, **p<0.01, and ***p<0.001). (C) Heat-map representing the fold-change expression profiles of GlyRα subunits in WBCs across different inflammatory conditions. The control group was normalized to 1, presented as green and red for upregulation. Distinct clustering patterns highlighted the strong upregulation of GlyRα subunits under neuroinflammation.

Article Snippet: After blocking with 5% normal goat serum (NGS) for 1 hour, the slides and coverslips were stained with primary antibodies targeting human and mouse GlyRα1 (1:500, #NB300-113, Novus Bio, Littleton, CO), GlyRα3 (1:100, #75-417, Neuromab, UC Davis/NIH), CD11b (1:100, #Sc-1186, Santa Cruz Biotechnology), CD-68 (1:100, #Sc-20060, Santa Cruz Biotechnology), and Iba1 (1:400, #MABN92, Millipore Sigma) ( ) overnight at 4°C.

Techniques: Immunofluorescence, Staining, Control, Expressing, Activation Assay

Journal: Frontiers in Immunology

Article Title: Glycine receptors in circulating white blood cells regulated by neuroinflammation

doi: 10.3389/fimmu.2026.1749275

Figure Lengend Snippet: Identification and localization of GlyRα subunits in human PBMCs and CD14+ monocytes. (A) Relative mRNA expression of GlyRα1, α2, and α3 in CD14+ monocytes (white bars) and PBMCs (gray bars), normalized to β2-microglobulin housekeeping gene. GlyRα1 showed the highest expression, followed by GlyRα2 and α3. Data are presented as mean ± SEM (B) Quantification of PBMCs expressing 55.21 ± 2.8% of GlyRα1/2+ cells and 46.87 ± 3.4% of GlyRα3+ cells, with 43.12 ± 2.2% of GlyRα1/2+/GlyRα3+ PBMCs. GlyRα1/2+ cells were significantly more numerous than dual-positive cells (p=0.015*). Statistical significance was assessed using one-way ANOVA; bars depict mean ± SEM; *p<0.05. (C) Representative immunofluorescence images of PBMCs showing GlyRα1/2 (red), GlyRα3 (green), and nuclei stained with DAPI (blue, arrowhead). The merged panel shows the co-localization of GlyRα1/2 and GlyRα3, with a few cells negative for GlyRs.

Article Snippet: After blocking with 5% normal goat serum (NGS) for 1 hour, the slides and coverslips were stained with primary antibodies targeting human and mouse GlyRα1 (1:500, #NB300-113, Novus Bio, Littleton, CO), GlyRα3 (1:100, #75-417, Neuromab, UC Davis/NIH), CD11b (1:100, #Sc-1186, Santa Cruz Biotechnology), CD-68 (1:100, #Sc-20060, Santa Cruz Biotechnology), and Iba1 (1:400, #MABN92, Millipore Sigma) ( ) overnight at 4°C.

Techniques: Expressing, Immunofluorescence, Staining

Journal: Frontiers in Immunology

Article Title: Glycine receptors in circulating white blood cells regulated by neuroinflammation

doi: 10.3389/fimmu.2026.1749275

Figure Lengend Snippet: Identification of GlyRα subunits in mouse brain, spleen, bone marrow, and blood. Normalized mRNA expression levels of GlyRα1, α2, and α3 in mouse brain (A) , spleen (C) , bone marrow (BM; E ), and blood (G) show the highest expression of GlyRα2, followed by GlyRα3 and α1. Statistical significance was determined using one-way ANOVA followed by Tukey’s post hoc test. Data are shown as mean ± SEM; p<0.01** (n=4-6 mice per group). (B) Representative immunofluorescence images of brain sections stained for GlyRα1/2 ( red ), GlyRα3 ( green ), and nuclei (DAPI, blue ). GlyRα1/2 and α3 signals show co-expression within neurons of the hippocampus. (D) Spleen sections display strong GlyRα1/2 (red) staining in both red (arrow) and white pulps (arrow head), whereas GlyRα3 (green) staining is localized to the macrophage zone of red pulp. (F, H) Immunostaining of BM and WBCs shows strong membrane expression of GlyRα1/2 and α3. GlyRα3 staining marks distinct cell populations distributed in the cytoplasm and membrane. The merged image shows GlyRα1/2 and α3 co-localization, indicating heterogeneity in receptor subunit expression among different cell BM and WBC subsets.

Article Snippet: After blocking with 5% normal goat serum (NGS) for 1 hour, the slides and coverslips were stained with primary antibodies targeting human and mouse GlyRα1 (1:500, #NB300-113, Novus Bio, Littleton, CO), GlyRα3 (1:100, #75-417, Neuromab, UC Davis/NIH), CD11b (1:100, #Sc-1186, Santa Cruz Biotechnology), CD-68 (1:100, #Sc-20060, Santa Cruz Biotechnology), and Iba1 (1:400, #MABN92, Millipore Sigma) ( ) overnight at 4°C.

Techniques: Expressing, Immunofluorescence, Staining, Immunostaining, Membrane

Journal: Frontiers in Immunology

Article Title: Glycine receptors in circulating white blood cells regulated by neuroinflammation

doi: 10.3389/fimmu.2026.1749275

Figure Lengend Snippet: GlyRα subunit response to LPS-treated monocytes and neutrophils (A) Experimental workflow showing BM cells from C57BL/6 mice were differentiated into macrophages (BMMs), treated with LPS (50 and 100 ng/mL), and analyzed by qRT-PCR and immunofluorescence. (B) LPS-induced dose-dependent increases in IL-1β and TNFα expression with IL-2 downregulation, confirming macrophage activation. Data was calculated using the 2 ^-ΔΔCt method (mean ± SEM) and analyzed using one-way ANOVA (*p<0.05, **p<0.01, and ***p<0.001). (C, D) Representative immunofluorescence images showing expression and co-localization of GlyRα1/2 and 3 ( red ) with CD11b+ monocytes/neutrophils ( green ). The enhanced GlyRα1/2 and α3 immunoreactivity in LPS-treated CD11b+ cells indicates activation-dependent upregulation. (E, F) Representative immunofluorescence images showing staining and colocalization of the macrophage marker CD68 ( green ) with GlyRα1/2 (E) and α3 ( F , red ) in control and LPS-treated BMMs. During inflammation, a noticeable increase in GlyRα1/2 and α3 fluorescence intensity was observed at the cell periphery, co-localizing with CD68, indicating possible increased receptor surface accumulation or transcriptional upregulation upon activation.

Article Snippet: After blocking with 5% normal goat serum (NGS) for 1 hour, the slides and coverslips were stained with primary antibodies targeting human and mouse GlyRα1 (1:500, #NB300-113, Novus Bio, Littleton, CO), GlyRα3 (1:100, #75-417, Neuromab, UC Davis/NIH), CD11b (1:100, #Sc-1186, Santa Cruz Biotechnology), CD-68 (1:100, #Sc-20060, Santa Cruz Biotechnology), and Iba1 (1:400, #MABN92, Millipore Sigma) ( ) overnight at 4°C.

Techniques: Quantitative RT-PCR, Immunofluorescence, Expressing, Activation Assay, Staining, Marker, Control, Fluorescence

Journal: Frontiers in Immunology

Article Title: Glycine receptors in circulating white blood cells regulated by neuroinflammation

doi: 10.3389/fimmu.2026.1749275

Figure Lengend Snippet: Neuronal GlyR response to systemic and neuroinflammation in mice. (A) Fold change in mRNA expression of GlyRα1, α2, and α3 in mouse brains (n=4-6 mice per group). Neuroinflammation induced upregulation of GlyRα1 and α3, with no change in GlyRα2 expression. Data are shown as mean ± SEM, calculated using the 2 ^-ΔΔCt method. The experiment was performed twice in duplicate. Statistical significance was determined using one-way ANOVA, with *p<0.05, **p<0.01, and ***p<0.001. (B) Heat map displaying relative fold-change expression of GlyRα subunits in control, systemic, and neuroinflammatory brain samples. Control values were normalized to 1, with fold changes represented by the color scale (green = baseline/control, red = upregulation). Clustering showed marked upregulation of GlyRα1 and α3 during neuroinflammation.

Article Snippet: After blocking with 5% normal goat serum (NGS) for 1 hour, the slides and coverslips were stained with primary antibodies targeting human and mouse GlyRα1 (1:500, #NB300-113, Novus Bio, Littleton, CO), GlyRα3 (1:100, #75-417, Neuromab, UC Davis/NIH), CD11b (1:100, #Sc-1186, Santa Cruz Biotechnology), CD-68 (1:100, #Sc-20060, Santa Cruz Biotechnology), and Iba1 (1:400, #MABN92, Millipore Sigma) ( ) overnight at 4°C.

Techniques: Expressing, Control

Journal: Frontiers in Immunology

Article Title: Glycine receptors in circulating white blood cells regulated by neuroinflammation

doi: 10.3389/fimmu.2026.1749275

Figure Lengend Snippet: Non-neuronal (spleen and BM cells) GlyRα subunit expression during systemic and neuroinflammation. (A) Spleen tissue (n=4-6 mice per group) showing significant upregulation in GlyRα1, α2, and α3 under neuroinflammatory conditions, while remaining unchanged during systemic inflammation. This suggests enhanced splenic immune activation in response to central neuroinflammation. (B) BM cells (n=4-6 mie per group) displayed increased GlyRα1 and α2 expression during neuroinflammation, whereas systemic inflammation reduced the expression of all three GlyRα subunits. Data are presented as mean ± SEM using the 2 ^-ΔΔCt method. Experiments were performed twice in duplicate. Statistical significance was determined by one-way ANOVA followed by Tukey’s post hoc test (*p<0.05, **p<0.01, and ***p<0.001). (C, D) Heat maps illustrated the relative fold-change expression profiles of GlyRα1, α2, and α3 in the spleen (C) and BM cells (D) . Control samples were assigned a baseline value of 1, with fold-change values (green = baseline/control, red = upregulation). The heat maps reveal distinct upregulation patterns of GlyRα subunits in both tissues, with stronger expression observed during neuroinflammatory conditions.

Article Snippet: After blocking with 5% normal goat serum (NGS) for 1 hour, the slides and coverslips were stained with primary antibodies targeting human and mouse GlyRα1 (1:500, #NB300-113, Novus Bio, Littleton, CO), GlyRα3 (1:100, #75-417, Neuromab, UC Davis/NIH), CD11b (1:100, #Sc-1186, Santa Cruz Biotechnology), CD-68 (1:100, #Sc-20060, Santa Cruz Biotechnology), and Iba1 (1:400, #MABN92, Millipore Sigma) ( ) overnight at 4°C.

Techniques: Expressing, Activation Assay, Control

Journal: Frontiers in Immunology

Article Title: Glycine receptors in circulating white blood cells regulated by neuroinflammation

doi: 10.3389/fimmu.2026.1749275

Figure Lengend Snippet: Blood-based GlyR response under different inflammatory conditions. (A) Representative immunofluorescence images showing GlyRα1/2 (red), GlyRα3 (green), and DAPI (blue) staining in WBCs from control, systemic- and neuroinflammation groups. Scale bar: 10µm (B) . Fold-change expression of GlyRα1, α2, and α3 in circulating WBCs (n=4-6 mice per group) showed that neuroinflammation significantly upregulated all three GlyRα subunits, while systemic inflammation selectively increased GlyRα1 and α2 expression. This suggests enhanced peripheral glycinergic activation in response to neuroinflammation. The experiment was repeated twice in duplicates. Statistical significance was assessed by one-way ANOVA followed by Tukey’s post hoc test (*p<0.05, **p<0.01, and ***p<0.001). (C) Heat-map representing the fold-change expression profiles of GlyRα subunits in WBCs across different inflammatory conditions. The control group was normalized to 1, presented as green and red for upregulation. Distinct clustering patterns highlighted the strong upregulation of GlyRα subunits under neuroinflammation.

Article Snippet: After blocking with 5% normal goat serum (NGS) for 1 hour, the slides and coverslips were stained with primary antibodies targeting human and mouse GlyRα1 (1:500, #NB300-113, Novus Bio, Littleton, CO), GlyRα3 (1:100, #75-417, Neuromab, UC Davis/NIH), CD11b (1:100, #Sc-1186, Santa Cruz Biotechnology), CD-68 (1:100, #Sc-20060, Santa Cruz Biotechnology), and Iba1 (1:400, #MABN92, Millipore Sigma) ( ) overnight at 4°C.

Techniques: Immunofluorescence, Staining, Control, Expressing, Activation Assay

Journal: Frontiers in Immunology

Article Title: Glycine receptors in circulating white blood cells regulated by neuroinflammation

doi: 10.3389/fimmu.2026.1749275

Figure Lengend Snippet: Blood-based biomarker function of GlyRs during neuroinflammation. (A) Representative immunofluorescence images of the left hemisphere cerebral cortex showing Iba1+ microglia (red) at day 0 (control), day 2, and day 5 following intracerebral neuroinflammation. Nuclei were counterstained with DAPI (blue). Iba1 immunoreactivity is markedly increased on day 2, indicating robust microglial activation. By day 5, Iba1+ microglial density partially decreases, suggesting resolution of the acute inflammatory response. Scale bar: 100 µm. (B) Quantification of Iba1+ microglial density (percentage index) in cortical sections confirms peak microglial activation at day 2, corresponding to the acute phase of neuroinflammation, with partial resolution by day 5. (C–E) GlyRα1, α2, and α3 expression in blood shows significant upregulation at day 2, followed by a decline to or below baseline levels by day 5. (F–H) Analysis of brain Iba1 (black lines) and blood GlyRα subunit (colored lines) expression during neuroinflammation reveals a coordinated central-peripheral association. (F) Iba1 and GlyRα1 elevation on day 2 reflects synchronized central-peripheral immune activation. By day 5, brain Iba1 remains moderately elevated while blood GlyRα1 declines below baseline, indicating rapid systemic resolution. (G, H) GlyRα2 and α3 exhibit similar transient upregulation at day 2, inversely correlating with the partial resolution of brain Iba1 by day 5, demonstrating a temporary reciprocal modulation between cortical microglia and peripheral GlyRα subunits. Data are presented as fold change (mean ± SEM), calculated using the 2 ^-ΔΔCt method. Experiments were performed twice in duplicate. Statistical significance was determined by one-way ANOVA followed by Tukey’s post hoc test (*p<0.05, **p<0.01, and ***p<0.001).

Article Snippet: After blocking with 5% normal goat serum (NGS) for 1 hour, the slides and coverslips were stained with primary antibodies targeting human and mouse GlyRα1 (1:500, #NB300-113, Novus Bio, Littleton, CO), GlyRα3 (1:100, #75-417, Neuromab, UC Davis/NIH), CD11b (1:100, #Sc-1186, Santa Cruz Biotechnology), CD-68 (1:100, #Sc-20060, Santa Cruz Biotechnology), and Iba1 (1:400, #MABN92, Millipore Sigma) ( ) overnight at 4°C.

Techniques: Biomarker Discovery, Immunofluorescence, Control, Activation Assay, Expressing