rabbit anti igfbp2  (Bioss)

Journal: Nature Communications

Article Title: Disease-relevant upregulation of P2Y 1 receptor in astrocytes enhances neuronal excitability via IGFBP2

doi: 10.1038/s41467-024-50190-7

Figure Lengend Snippet: A , B Immunohistochemical analysis of IGFBP2 expression in the hippocampus. GFAP was used as an astrocytic marker. IGFBP2 signals that were colocalized with GFAP were quantified. Control, n = 293 ROIs from 15FOVs, 3 mice; AstroP2Y1OE, n = 331 ROIs from 12 FOVs, 3 mice (two-sided two-sample t-test). Data are presented as mean ± s.d. C Representative traces of EFS-evoked neuronal Ca 2+ signals (magenta) and astrocytic Ca 2+ signals (green). Compared with control IgG (n = 20 ROIs from 3 slices, 3 mice), IGFBP2 antibodies reduced neuronal Ca 2+ signals without affecting astrocytic Ca 2+ signals in AstroP2Y1OE mice (n = 40 ROIs from 6 slices, 3 mice) (two-sided two-sample t-test). Data are presented as mean ± s.d. D IGFBP2 at 10 ng/mL (1 h at room temperature) enhanced EFS-evoked neuronal Ca 2+ signals in the soma. ACSF (control), n = 42 cells from 5 slices, 3 mice; IGFBP2, n = 61 cells from 6 slices, 3 mice (two-sided two-sample t-test). Data are presented as mean ± s.d. E Schematic diagram for the AAV-mediated knockdown of Igfbp2 in astrocytes. The schematic diagram in Fig. was created with BioRender.com released under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International license. F Traces of neuronal Ca 2+ signals in the soma of AstroP2Y1OE mice with or without Igfbp2 knockdown in astrocytes. Control, n = 55 cells from 9 slices, 4 mice; Igfbp2 gRNA, n = 33 cells from 5 slices, 3 mice (two-sided Mann–Whitney U test). Data are presented as mean ± s.d. Source data are provided as a Source Data file.

Article Snippet: Slices were washed three times with PBS before being treated with 0.5% Triton-X100/10% normal goat serum (VectorLaboratories Cat# S-1000-20) for 1 h. After three washes with PBS, sections were incubated for 48 h with the following primary antibodies: rabbit anti-IGFBP2 antibody (1:500, Abcam, RRID:AB_2938998), mouse anti-IGFBP2 antibody (1:100, Santa Cruz Biotechnology, Cat# sc-515134), rat anti-GFAP antibody (1:2000, Thermo Fisher Scientific; RRID: AB_2532994), rabbit anti-P2Y1R antibody (1:250, Alomone Labs; RRID: AB_2040070), rabbit anti-SOX9 antibody (1:1000, Millipore; RRID: AB_2239761), rat anti-mCherry (16D7) (1:2000, Thermo Fisher Scientific, RRID: AB_2536611), mouse anti-hemagglutinin (HA) antibody (1:500, BioLegend, RRID: AB_2565336), guinea pig anti-GLT-1 antibody (1:1000, Sigma-Aldrich, RRID:AB_90949), and rabbit anti-GLAST antibody (1:200, Nittobo Medical, MSFR102120).

Techniques: Immunohistochemical staining, Expressing, Marker, Control, Knockdown, MANN-WHITNEY

Journal: Nature Communications

Article Title: Disease-relevant upregulation of P2Y 1 receptor in astrocytes enhances neuronal excitability via IGFBP2

doi: 10.1038/s41467-024-50190-7

Figure Lengend Snippet: A Representative images of GFAP, P2Y1R, and IGFBP2 immunohistochemistry in the hippocampal CA1 region. The images were taken from the side contralateral to the kainic acid injection. Note that P2Y1R and IGFBP2 were upregulated in reactive astrocytes. These astrocytes were similarly distributed in the hippocampal CA1 region. The bar graph shows a summary of P2Y1R- and IGFBP2-positive GFAP-positive astrocytes. n = 6–7 slices from 3 mice for each group (two-sided Mann–Whitney U test). Data are presented as mean ± s.d. B Immunohistochemistry for IGFBP2 and P2Y1R in astrocytes was performed in brains 3 days after MCAO. Expression of IGFBP2 and P2Y1R appeared in GFAP-positive astrocytes in the striatum ipsilateral to MCAO. There was no obvious expression of either P2Y1R or IGFBP2 in the side contralateral to MCAO. The bar graph shows a summary of P2Y1R- and IGFBP2-positive GFAP-positive astrocytes. n = 6 slices from 4 mice (two-sided Mann–Whitney U test). Data are presented as mean ± s.d. Source data are provided as a Source Data file.

Article Snippet: Slices were washed three times with PBS before being treated with 0.5% Triton-X100/10% normal goat serum (VectorLaboratories Cat# S-1000-20) for 1 h. After three washes with PBS, sections were incubated for 48 h with the following primary antibodies: rabbit anti-IGFBP2 antibody (1:500, Abcam, RRID:AB_2938998), mouse anti-IGFBP2 antibody (1:100, Santa Cruz Biotechnology, Cat# sc-515134), rat anti-GFAP antibody (1:2000, Thermo Fisher Scientific; RRID: AB_2532994), rabbit anti-P2Y1R antibody (1:250, Alomone Labs; RRID: AB_2040070), rabbit anti-SOX9 antibody (1:1000, Millipore; RRID: AB_2239761), rat anti-mCherry (16D7) (1:2000, Thermo Fisher Scientific, RRID: AB_2536611), mouse anti-hemagglutinin (HA) antibody (1:500, BioLegend, RRID: AB_2565336), guinea pig anti-GLT-1 antibody (1:1000, Sigma-Aldrich, RRID:AB_90949), and rabbit anti-GLAST antibody (1:200, Nittobo Medical, MSFR102120).

Techniques: Immunohistochemistry, Injection, MANN-WHITNEY, Expressing

Journal: Nature Communications

Article Title: Disease-relevant upregulation of P2Y 1 receptor in astrocytes enhances neuronal excitability via IGFBP2

doi: 10.1038/s41467-024-50190-7

Figure Lengend Snippet: Astrocytic expression of P2ry1/P2RY1 and Igfbp2/IGFBP2 in neurological disease models in publicly available datasets

Article Snippet: Slices were washed three times with PBS before being treated with 0.5% Triton-X100/10% normal goat serum (VectorLaboratories Cat# S-1000-20) for 1 h. After three washes with PBS, sections were incubated for 48 h with the following primary antibodies: rabbit anti-IGFBP2 antibody (1:500, Abcam, RRID:AB_2938998), mouse anti-IGFBP2 antibody (1:100, Santa Cruz Biotechnology, Cat# sc-515134), rat anti-GFAP antibody (1:2000, Thermo Fisher Scientific; RRID: AB_2532994), rabbit anti-P2Y1R antibody (1:250, Alomone Labs; RRID: AB_2040070), rabbit anti-SOX9 antibody (1:1000, Millipore; RRID: AB_2239761), rat anti-mCherry (16D7) (1:2000, Thermo Fisher Scientific, RRID: AB_2536611), mouse anti-hemagglutinin (HA) antibody (1:500, BioLegend, RRID: AB_2565336), guinea pig anti-GLT-1 antibody (1:1000, Sigma-Aldrich, RRID:AB_90949), and rabbit anti-GLAST antibody (1:200, Nittobo Medical, MSFR102120).

Techniques: Expressing, Comparison, Muscles

Journal: Nature Communications

Article Title: Disease-relevant upregulation of P2Y 1 receptor in astrocytes enhances neuronal excitability via IGFBP2

doi: 10.1038/s41467-024-50190-7

Figure Lengend Snippet: P2Y1R is upregulated in reactive astrocytes in several neurological disorders such as epilepsy, stroke, Alzheimer’s disease, and traumatic brain injury. The increase in P2Y1R signaling in astrocytes leads to neuronal hyperexcitability by increased expression of IGFBP2, which acts as an excitatory signal on neurons. Our data indicate that the functional phenotype of reactive astrocytes with upregulated P2Y1R-IGFBP2 signaling is shared in several neurological diseases including epilepsy and stroke and may contribute to neuronal hyperexcitability.

Article Snippet: Slices were washed three times with PBS before being treated with 0.5% Triton-X100/10% normal goat serum (VectorLaboratories Cat# S-1000-20) for 1 h. After three washes with PBS, sections were incubated for 48 h with the following primary antibodies: rabbit anti-IGFBP2 antibody (1:500, Abcam, RRID:AB_2938998), mouse anti-IGFBP2 antibody (1:100, Santa Cruz Biotechnology, Cat# sc-515134), rat anti-GFAP antibody (1:2000, Thermo Fisher Scientific; RRID: AB_2532994), rabbit anti-P2Y1R antibody (1:250, Alomone Labs; RRID: AB_2040070), rabbit anti-SOX9 antibody (1:1000, Millipore; RRID: AB_2239761), rat anti-mCherry (16D7) (1:2000, Thermo Fisher Scientific, RRID: AB_2536611), mouse anti-hemagglutinin (HA) antibody (1:500, BioLegend, RRID: AB_2565336), guinea pig anti-GLT-1 antibody (1:1000, Sigma-Aldrich, RRID:AB_90949), and rabbit anti-GLAST antibody (1:200, Nittobo Medical, MSFR102120).

Techniques: Expressing, Functional Assay