Journal: Scientific Reports

Article Title: Functional NMDA receptors are expressed by human pulmonary artery smooth muscle cells

doi: 10.1038/s41598-021-87667-0

Figure Lengend Snippet: Functional NMDA receptors are localized on the surface of HPASMCs. Immunofluorescence staining was performed on non-permeabilized HPASMCs. Both GluN1 and GluN2 (2B and 2D) subunits were detected on the surface of HPASMCs ( A , B ). GluN1 co-localized with GluN2B ( A ) and GluN2D ( B ) on the surface of HPASMCs, respectively. Results are representative images taken from at least three separate experiments. Scale bar = 100 µM. ( C , D ) are representative scatter plots of each fluorescent pixel from confocal images with GluN1 green fluorescent intensity along the x -axis and GluN2 red fluorescent intensity along the y -axis. The shaded area in the upper right quadrant represents colocalized pixels above background with the associated Pearson correlation coefficient indicated for all colocalized pixels. ( E ) Representative traces for cultured HPASMCs in response to 100 μM NMDA and 10 μM glycine treatment. Cells were clamped at − 50 mV and whole cell recordings were performed. ( F ) NMDA and glycine treatment evoked an inward whole-cell current of 10.9 ± 1.7 pA (Mean ± SE, **p < 0.01, n = 16). ( G ) NMDA and glycine treatment evoked increased AUC compared with baseline (*p < 0.05, n = 16).

Article Snippet: After 3 washes in TBS, sections were permeabilized and pre-blocked followed by primary antibody incubation at the following concentrations: platelet endothelial cell adhesion molecule (PECAM-1) (ab28364, Abcam, Cambridge, MA) (1/150), GluN1 (556308, BD Pharmingen, San Jose, CA) (1/1000) (this reference contains characterization of the antibody against GluN1), GluN2A (AGC-002, Alomone, Jerusalem, Israel) (1/20,000), GluN2B (AGC-003, Alomone, Jerusalem, Israel) (1/3000), GluN2C (ab105146, Abcam, Cambridge, MA) (1/30,000) or GluN2D (ab186816, Abcam, Cambridge, MA) (1/10,000).

Techniques: Functional Assay, Immunofluorescence, Staining, Cell Culture

Journal: Scientific Reports

Article Title: Distribution and relative expression of vasoactive receptors on arteries

doi: 10.1038/s41598-020-72352-5

Figure Lengend Snippet: Vasoactive receptors and ligands assayed in this study.

Article Snippet: Membranes were blocked with 5% nonfat dry milk for 1 h at room temperature and incubated overnight at 4 °C with primary antibodies against ADRA2B (1:2000; AP17872PU-N, Origene) or GAPDH (1:5000; ab8245, Abcam).

Techniques:

Journal: Scientific Reports

Article Title: Distribution and relative expression of vasoactive receptors on arteries

doi: 10.1038/s41598-020-72352-5

Figure Lengend Snippet: ( a ) Gray values of six subtypes of alpha-adrenergic receptor (ADRA) in coronary, pulmonary, renal, mesenteric and peripheral arteries. ( b ) Gray values of six subtypes of ADRA in coronary artery. ( c ) Gray values of ADRA2B in five types of arteries. ( d ) Western blot of ADRA2B in coronary (C), peripheral (P), renal (R), and mesenteric artery (M). GADPH served as a loading control. ( e ) Immunohistochemistry against six subtypes of ADRA in myocardium. Sections were stained against alpha smooth muscle actin (alpha-SM) to locate arteries. Red arrows: artery with diameter < 50 μm. Bottom right corner: artery with diameter ≥ 50 μm. As a negative control, sections were incubated with phosphate-buffered saline instead of primary antibody. Data are mean ± SD, and P < 0.05 for Student’s t test was set as the significance threshold.

Article Snippet: Membranes were blocked with 5% nonfat dry milk for 1 h at room temperature and incubated overnight at 4 °C with primary antibodies against ADRA2B (1:2000; AP17872PU-N, Origene) or GAPDH (1:5000; ab8245, Abcam).

Techniques: Western Blot, Control, Immunohistochemistry, Staining, Negative Control, Incubation, Saline

Journal: Frontiers in Cardiovascular Medicine

Article Title: Serotonin transporter downregulation is associated with aortic stenosis, and early profibrotic remodeling is mitigated by pharmacological inhibition of HTR2B receptor

doi: 10.3389/fcvm.2026.1729078

Figure Lengend Snippet: Effects of AngII on human AVICs with SERT knockdown. (A) Gene expression of SERT, HTR2B, HTR2A in AngII-treated compared to Nontreated (NT) Ctrl AVICs. (B) SERT KD by siRNA lead to increased expression of HTR2A gene expression. (C) HTR2A and HTR2B expression with siSERT or siSERT combined with AngII treatment. (D) COL1A1, SPP1, RUNX2, TGFβ1 expression in response to AngII treatment in human AVICs compared to Nontreated group. (E) COL1A1, RUNX2, TGFβ1 expression treated with siSERT alone or siSERT combined with AngII treatment. All gene expression results were calculated by the 2 −ΔΔCT method, n ≥ 4 per group. Error bars indicate SEM. P-value vs. NT. *indicates p-value <0.1, **indicates p-value <0.05, and ***indicates p -value <0.01 by Student's t -test or one-way ANOVA with post-hoc Dunnett's test.

Article Snippet: A two-day immunohistochemistry protocol was used for HTR2B receptor (ProteinTech, 26408-1-AP).

Techniques: Knockdown, Gene Expression, Expressing

Journal: Frontiers in Cardiovascular Medicine

Article Title: Serotonin transporter downregulation is associated with aortic stenosis, and early profibrotic remodeling is mitigated by pharmacological inhibition of HTR2B receptor

doi: 10.3389/fcvm.2026.1729078

Figure Lengend Snippet: Phenotype of calcified AS aortic valve leaflets, includes SERT expression downregulation and HTR2B upregulation (A) representative echocardiography images of a patient with AS prior to surgical aortic valve replacement; LA, left atrium; AV, aortic valve; LV, left ventricle. Arrow 1 (top panel, white) indicates the thickened and calcified aortic valve. Arrow 2 (top panel, white) indicates AS jet visualized by Doppler ultrasound. (B) Resected aortic leaflet of human normal AV and stenotic AV (top). Representative topographic images generated of normal AV and stenotic AV by 3-Dimensional KEYENCE Laser profiler (bottom). Colors corresponding from high to low respectively: red-orange-yellow-green-blue. (C) Alizarin Red staining of normal AV samples from heart donors (left) and AS AV leaflets resected during surgery (right); f, zona fibrosa; s, zona spongiosa; v, zona ventricularis. Alizarin Red shows accumulation of calcium deposits (red/orange). The images were taken under brightfield microscope a 20× objective. (D) Representative immunofluorescence staining of SERT and the quantification of mean fluorescent intensity in normal AV and AS AV tissue (green). (E) Representative immunofluorescence staining of HTR2B receptor and the quantification of mean fluorescent intensity in normal and AS AV tissue (green). All immunofluorescent images were taken with a confocal microscope at 20× magnification. Error bars indicate SD. **Indicates p-value < 0.05 with unpaired Student's t -test.

Article Snippet: A two-day immunohistochemistry protocol was used for HTR2B receptor (ProteinTech, 26408-1-AP).

Techniques: Expressing, Generated, Staining, Microscopy, Immunofluorescence

Journal: Frontiers in Cardiovascular Medicine

Article Title: Serotonin transporter downregulation is associated with aortic stenosis, and early profibrotic remodeling is mitigated by pharmacological inhibition of HTR2B receptor

doi: 10.3389/fcvm.2026.1729078

Figure Lengend Snippet: Bulk RNA sequencing reveals the reversal of AngII-induced remodeling in mouse AVs via the treatment with HTR2B antagonist, LY272015 . (A) Volcano plot of Differentially Expressed Genes (DEGs) number, p-value <0.05 & |log2foldchange| > 1, of the AngII-treated group compared to the Ctrl group. (B) Heatmap showing distribution of DEGs among different samples. (C) Ingenuity Pathway Analysis showing top differentially regulated pathways (-log( p -value) > 0). (D) Volcano plot of Differentially Expressed Genes (DEGs) number, p-value < 0.05 & |log2foldchange| > 1, in the AngII+LY-treated group compared to the AngII group. (E) Heatmap showing distribution of DEGs among different samples of AngII+LY and AngII groups. (F) Ingenuity Pathway Analysis showing top differentially regulated pathways (-log( p -value) > 0) in the AngII+LY-treated group compared to the AngII group. Z-scores indicate the likelihood of pathway activation or inhibition. Positive Z-score indicates activated and Negative Z-score indicates inhibited pathways.

Article Snippet: A two-day immunohistochemistry protocol was used for HTR2B receptor (ProteinTech, 26408-1-AP).

Techniques: RNA Sequencing, Activation Assay, Inhibition

Journal: The Journal of Biological Chemistry

Article Title: A bispecific antibody designed to act as a NRP2/PLXNA1 agonist mimics anticancer activity of SEMA3F

doi: 10.1016/j.jbc.2025.111056

Figure Lengend Snippet: SEMA3F dimerizes surface NRP2 and PLXNA1, inhibits U-251 MG cell proliferation, and reduces p-AKT level and CCND1 gene expression . A , effects of SEMA3F on proliferation of various cancer cell lines. Cells were treated with full-length SEMA3F for 3 days, and cell viability was measured using CellTiter-Glo, normalized to untreated cells. B , schematic illustrates the full-length SEMA3F dimer, the furin processing site, and the resulting furin-processed SEMA3F (SEMA3F-p65). C , SEMA3F-p65 does not inhibit U-251 MG cell proliferation compared to nonprocessed SEMA3F dimer. D , the anti-NRP2 antibody (aNRP2-a2) blocks SEMA3F-mediated inhibition of U-251 MG cell proliferation. E , schematic illustrating the receptor dimerization assay. NanoLuc luciferase is split into Large BiT and Small BiT with low activity, and fused to the N termini of NRP2 and PLXNA1. SEMA3F induces NRP2-PLXNA1 dimerization, bringing Large BiT and Small BiT together to enhance luciferase activity through complementation. F , SEMA3F treatment induces NRP2-PLXNA1 dimerization. Left panel : Expi293F cells coexpressing Large_BiT-NRP2 and Small_BiT-PLXNA1 were treated with luciferase substrate and SEMA3F as indicated, and luminescence was recorded over time. Right panel : dimerization ratio changes following SEMA3F treatment. G , SEMA3F reduces p-AKT levels in U-251 MG cells in an NRP2-dependent manner. U-251 MG cells were treated as indicated for 30 min, and the p-AKT/AKT ratio was measured. H , SEM3F downregulates CCND1 expression in an NRP2-dependent manner. U-251 MG cells treated for 18 h. The expression of CCND1 was quantified using qPCR and normalized to untreated control cells. I , schematic representation of SEMA3F’s anti-proliferative mechanism in U-251 MG cells. NRP, neuropilin; aNRP2, anti-NRP2; PLXNA1, plexinA1; SEMA, semaphorin; p-AKT, phosphorylation of AKT; qPCR, quantitative PCR.

Article Snippet: Dimerization between cell surface NRP2 and PLXNA1 proteins were determined using a luciferase complementation assay as described previously ( ). cDNAs of NRP2 and PLXNA1 were obtained from OriGene and R&D Systems respectively (NRP2 C220706, PLXNA1 RDC0967).

Techniques: Gene Expression, Inhibition, Luciferase, Activity Assay, Expressing, Control, Phospho-proteomics, Real-time Polymerase Chain Reaction

Journal: The Journal of Biological Chemistry

Article Title: A bispecific antibody designed to act as a NRP2/PLXNA1 agonist mimics anticancer activity of SEMA3F

doi: 10.1016/j.jbc.2025.111056

Figure Lengend Snippet: Experimental workflow for the discovery of SEMA3F-mimetic bsAbs . MAb-targeting PLXNA1 (aPLXNA1) were generated and paired with anti-NRP2 antibodies to create bispecific antibodies (bsAbs). These bsAbs were initially evaluated in a PLXNA1-NRP2 dimerization assay to identify candidates that effectively promote PLXNA1–NRP2 interaction. Positive bsAbs were then assessed using a p-AKT assay, a qPCR assay for CCND1 expression, and a cell viability assay to determine which candidates mimic SEMA3F’s effects. Ultimately, one bsAb, P1943-Nb2cL, was identified as mimicking SEMA3F’s effects across all cell-based assays. aPLXNA1, anti-PLXNA1; NRP, neuropilin; PLXNA1, plexinA1; SEMA, semaphorin; p-AKT, phosphorylation of AKT; qPCR, quantitative PCR.

Article Snippet: Dimerization between cell surface NRP2 and PLXNA1 proteins were determined using a luciferase complementation assay as described previously ( ). cDNAs of NRP2 and PLXNA1 were obtained from OriGene and R&D Systems respectively (NRP2 C220706, PLXNA1 RDC0967).

Techniques: Generated, Expressing, Viability Assay, Phospho-proteomics, Real-time Polymerase Chain Reaction

Journal: The Journal of Biological Chemistry

Article Title: A bispecific antibody designed to act as a NRP2/PLXNA1 agonist mimics anticancer activity of SEMA3F

doi: 10.1016/j.jbc.2025.111056

Figure Lengend Snippet: Identification of PLXNA1-NRP2 bsAb with SEMA3F-mimicking mechanism and activities . A , schematic illustrating structural format of PLXNA1-NRP2 bsAbs. The bsAbs are based on a human IGG4 backbone, with the light chain and heavy chain of the anti-NRP2 half linked by a 34 amino acid GS flexible peptide linker. The anti-NRP2 moiety contains a knob mutation, while the anti-PLXNA1 moiety features a hole mutation. B , screening for bsAbs that induce dimerization of cell surface PLXNA1 and NRP2. Each table cell represents a unique PLXNA1-NRP2 bsAb. bsAbs inducing a dimerization ratio change greater than 1.5 are considered positive hits and highlighted in orange . C , PLXNA1-NRP2 bsAbs that significantly reduced p-AKT level. D , PLXNA1-NRP2 bsAbs that significantly reduced CCND1 expressing in qPCR assay. E , PLXNA1-NRP2 bsAbs that significantly inhibited proliferation of U-251 MG cells. All data are presented as the mean ± SEM from three experiments. aPLXNA1, anti-PLXNA1; bsAb, bispecific antibody; NRP, neuropilin; p-AKT, phosphorylation of AKT; PLXNA1, plexinA1; qPCR, quantitative PCR; SEMA, semaphorin.

Article Snippet: Dimerization between cell surface NRP2 and PLXNA1 proteins were determined using a luciferase complementation assay as described previously ( ). cDNAs of NRP2 and PLXNA1 were obtained from OriGene and R&D Systems respectively (NRP2 C220706, PLXNA1 RDC0967).

Techniques: Mutagenesis, Expressing, Phospho-proteomics, Real-time Polymerase Chain Reaction

Journal: The Journal of Biological Chemistry

Article Title: A bispecific antibody designed to act as a NRP2/PLXNA1 agonist mimics anticancer activity of SEMA3F

doi: 10.1016/j.jbc.2025.111056

Figure Lengend Snippet: Characterization of the binding mechanism of aPLXNA1-19-43 Fab to PLXNA1 . A , epitope mapping of aPLXNA1-19-43 Fab to PLXNA1. ELISA was used to evaluate binding of aPLXNA1-19-43 Fab to various PLXNA1 fragments. Results are indicated by ELISA readings, with positive (+) binding shown in red for readings >0.1 and negative (−) binding for readings <0.1. B , determination of binding affinity of aPLXNA1-19-43 Fab to PLXNA1 ECD and LBD. Representative sensorgrams from triplicate experiments are shown. The Fab was immobilized using biosensor tips coated with anti-mouse kappa antibody and subsequently exposed to a concentration series of PLXNA1-ECD or PLXNA1-LBD ( black and gray lines ). Data were fitted to a 1:1 binding model ( red lines ) to calculate binding constants. C , cryo-EM density map of the aPLXNA1-19-43 Fab in complex with PLXNA1-LBD. The 2:2 dimeric complex is displayed, with one subunit colored as follows: PLXNA1-LBD in blue , Fab heavy chain in cyan , and Fab light chain in red . D , schematic binding model of aPLXNA1-19-43 Fab with PLXNA1. The critical residues involved in binding are enlarged in the inserted panels . E , identification of critical residues on PLXNA1 that are involved in the binding of aPLXNA1-19-43 Fab. The Fab is shown to bind to both the SEMA and PSI domains of PLXNA1, with key interactions highlighted. F , binding model of P1943-Nb2cL to the PLXNA1–NRP2–SEMA3F complex. G , schematic representation of the binding mechanism between P1943-Nb2cL and the PLXNA1–NRP2–SEMA3F complex, providing a simplified visual overview. aPLXNA1, anti-PLXNA1; ECD, extracellular domain; Fab, fragment of antigen binding; LBD, ligand-binding domain; NRP, neuropilin; PLXNA1, plexinA1; PSI, plexin-semaphorin-integrin; SEMA, semaphorin.

Article Snippet: Dimerization between cell surface NRP2 and PLXNA1 proteins were determined using a luciferase complementation assay as described previously ( ). cDNAs of NRP2 and PLXNA1 were obtained from OriGene and R&D Systems respectively (NRP2 C220706, PLXNA1 RDC0967).

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Concentration Assay, Cryo-EM Sample Prep, Ligand Binding Assay

Journal: International Journal of Molecular Sciences

Article Title: TRPV2: A Key Player in Myelination Disorders of the Central Nervous System

doi: 10.3390/ijms23073617

Figure Lengend Snippet: TRPV2 interaction with Opalin, NTM and PLP1 in vitro during basal and inflammatory conditions. ( A ) Rat TRPV2 FL protein and rat ARD of TRPV2 were immobilized on membranes and incubated with increasing concentrations of rat protein brain lysates (0–10 µg). Opalin, NTM and PLP1 were detected by immunoblot. ( B ) Immunocytochemistry in mouse mixed glial primary cultures shows TRPV2 (green), Opalin (red) and DAPI (blue). TRPV2-Opalin colocalization is shown as yellow signal (merge) and both proteins are co-expressed in some cells (arrowheads). Expression of these proteins is mainly found in the membrane. ( C ) TRPV2–Opalin colocalization expressed as Pearson’s coefficient in basal and inflammatory conditions. Both proteins show a high degree of colocalization. ( D ) TRPV2 colocalization with Opalin, expressed as Mander’s coefficient M2 (fraction of TRPV2 signal overlapping Opalin) in basal and inflammatory conditions. Both proteins show a high degree of colocalization. Results were expressed as mean ± SEM. Statistical analysis of the results were performed with an unpaired Student’s t -test ( p < 0.05 is statistically significant). Scale bar = 20 µm.

Article Snippet: TRPV2 , Rabbit , 1:200 , ACC-032, Alomone , Biotinylated horse anti-rabbit , 1:500 , BA-1100, Vector Laboratories.

Techniques: In Vitro, Incubation, Western Blot, Immunocytochemistry, Expressing

Journal: International Journal of Molecular Sciences

Article Title: TRPV2: A Key Player in Myelination Disorders of the Central Nervous System

doi: 10.3390/ijms23073617

Figure Lengend Snippet: TRPV2 expression in mixed glia cultures of mice after pro-inflammatory (LPS), anti-inflammatory (IL-4) and demyelinating (LPC) treatments. ( A – I ) Double immunohistochemical staining allowed for the determination of whether TRPV2 was expressed in microglia ( A – C ), astrocytes ( D – F ) or oligodendrocyte cells ( G – I ). TRPV2 was highly expressed in the cell body of some microglia (full arrowheads) but not all of them (empty arrowheads) in control conditions, and TRPV2 was spread afterwards through the cell body of activated microglia cells, with an expression pattern highly coincident with CD68+ lysosomes (arrows). While no expression of TRPV2 was found in astrocytes (empty arrowheads), OPCs showed either low or absent expression in control conditions (full and empty arrowheads, respectively) and increased expression throughout the cell body (arrows) and the principal soma (arrowheads) after both pro-inflammatory and anti-inflammatory treatments. ( J , K ) Quantification of TRPV2 protein (ng/mL) by ELISA in secondary mixed glial cultures in control conditions, and after LPS and IL-4 treatments ( J ) and in quaternary mixed glial cultures, containing myelin-binding protein (MBP)+ oligodendrocytes, in demyelinating conditions ( K ). Determination of TRPV2 showed a significant decrease in TRPV2 after LPS and LPC treatments ( J , K ), and an increase after IL-4 treatment ( p = 0.05) compared to control conditions ( J ). ( L , M ) Quantification of NO secondary mixed glial cultures in control conditions, and after LPS and IL-4 treatments ( L ) and in quaternary mixed glial cultures in demyelinating conditions ( M ). An increase in NO concentration was observed in cell cultures after LPS treatment compared to control conditions, while a significant decrease was observed after IL-4 treatment. After LPC treatment, NO remained stable compared to the control. Statistical analysis was performed by one-way ANOVA followed by Dunnett’s multiple comparison compared to the control in the LPS and IL-4 treated cultures (* p < 0.05, ** p < 0.01), and a paired Student’s t -test for LPC treatment (** p < 0.01). Scale bar ( A – I ) = 25 µm.

Article Snippet: TRPV2 , Rabbit , 1:200 , ACC-032, Alomone , Biotinylated horse anti-rabbit , 1:500 , BA-1100, Vector Laboratories.

Techniques: Expressing, Immunohistochemical staining, Staining, Enzyme-linked Immunosorbent Assay, Binding Assay, Concentration Assay

Journal: International Journal of Molecular Sciences

Article Title: TRPV2: A Key Player in Myelination Disorders of the Central Nervous System

doi: 10.3390/ijms23073617

Figure Lengend Snippet: TRPV2, Opalin and MSRA expression in spinal cord of wild-type (WT) and hypomyelination jimpy mice. Immunohistochemical staining against MBP ( A , B ), TRPV2 ( D , E ), Opalin ( G , H ) and MSRA ( J , K ) were performed in spinal cord sections of 21-day-old WT and jimpy mice, and immunoreactivity was quantified and analyzed as Area x Intensity. Results show that MBP was importantly reduced in both GM ( a , b ) and WM ( a’ , b’ ) of jimpy mice compared to WT ( A – C ). TRPV2 expression was mainly located in neurons in GM ( d’ ) and glial cells, putatively oligodendrocytes, and in WM ( d ) of WT. A significant decrease in TRPV2 expression in the spinal cord of jimpy mice was observed compared to WT ( E , F , e , e’ ). Opalin expression was importantly found in WM of WT mice ( G , g ), and to a lesser extent also in GM ( g’ ), but showed a marked decrease in WM and GM of jimpy mice ( H – I , h , h’ ). MSRA was the only molecule that did not suffer an important reduction of its expression in both WM ( j , k ) and GM ( j’ , k’ ) in jimpy mice ( L ). Results were expressed as mean ± SEM. Statistical analysis of the results were performed with an unpaired Student’s t -test (** p < 0.01, *** p < 0.001). Scale bar ( A – K ) = 50 µm; ( a – k , a’ – k’ ) = 20 µm.

Article Snippet: TRPV2 , Rabbit , 1:200 , ACC-032, Alomone , Biotinylated horse anti-rabbit , 1:500 , BA-1100, Vector Laboratories.

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: International Journal of Molecular Sciences

Article Title: TRPV2: A Key Player in Myelination Disorders of the Central Nervous System

doi: 10.3390/ijms23073617

Figure Lengend Snippet: TRPV2 and MSRA expression in WM of cuprizone-induced demyelination and remyelination in mice. ( A – I ) Representative images of immunohistochemical staining directed against MBP, TRPV2 and MSRA in the CC of control mice ( A , D , G ) and cuprizone-intoxicated mice after demyelination ( B , E , H , 5 weeks of treatment) and during remyelination ( C , F , I , 5 weeks of treatment + 1 week of normal diet). ( A – C ) MBP immunostaining shows the myelinated CC of WT mice in basal conditions ( A ). At 5 weeks of cuprizone treatment, the CC is largely demyelinated ( B ) with a myelination-resistant area (*), and after one week of normal diet, the CC is mostly remyelinated ( C ). Control mice show undetectable levels of TRPV2 and MSRA in the CC ( D and G ). After demyelination, variably low and high levels of TRPV2 were observed in cell bodies (empty arrowheads and arrowheads, respectively in E , F ). High levels of TRPV2 were clearly observed at remyelination (inset in F ). During remyelination, most TRPV2+ cells co-expressed APC (full arrowheads in L ), a marker for mature oligodendrocytes. In the CC area resistant to demyelination (* in E , F ), TRPV2+ cells showed high levels of expression in both conditions. An increase in MSRA levels of expression were also observed in cell bodies after demyelination and remyelination (arrowheads in H and I ). In demyelination, most cells expressed low levels of MSRA in the CC (inset in H ), while during remyelination, a lower number of MSRA+ cells expressing higher levels of MSRA were observed (inset in I ). ( J – K ) Quantification of TRPV2 and MSRA expression after cuprizone treatment. ( J ) Quantitative analysis of TRPV2 immunoreactivity in the CC confirmed the progressive increase of TRPV2 expression in demyelinating and remyelinating conditions compared to control mice, which was statistically significant in the latter. ( K ) Quantitative analysis of MSRA immunoreactivity in the CC showed that the increase of MSRA expression during demyelination was statistically significant when compared to control mice. Data are shown as ± SEM. Statistical analysis was performed by one-way ANOVA followed by Tukey’s multiple comparison test (** p < 0.01). Scale bar ( A – I ) = 50 μm; ( L ) = 100 μm.

Article Snippet: TRPV2 , Rabbit , 1:200 , ACC-032, Alomone , Biotinylated horse anti-rabbit , 1:500 , BA-1100, Vector Laboratories.

Techniques: Expressing, Immunohistochemical staining, Staining, Immunostaining, Marker

Journal: International Journal of Molecular Sciences

Article Title: TRPV2: A Key Player in Myelination Disorders of the Central Nervous System

doi: 10.3390/ijms23073617

Figure Lengend Snippet: TRPV2 expression in spinal cord samples from WT and EAE mice. ( A ) Immunohistochemical staining of TRPV2 in spinal cord of control and EAE-induced mice in the onset, peak and in the chronic phase, during remission. In control conditions, TRPV2 is expressed in neurons in GM (inset, arrows), and in oligodendrocytes in WM (inset, arrowheads). After EAE induction, an increase in TRPV2 expression is mainly located in inflammatory focuses at the peak clinical symptomatology in WM (empty arrowheads). ( B ) MSRA and TRPV2 protein expression in thoracic spinal cord samples from CFA and MOG mice (9, 14, 21 and 28 days post-immunization). ( B ) A significant increase in TRPV2 protein levels (#, p = 0.0014) is determined in MOG14 mice when compared with CFA ( p < 0.05) and when compared with MOG9 ( p < 0.05). A significant increase ( p < 0.05) in MSRA protein levels is observed in all groups of mice treated with MOG when compared with CFA mice, as well as between different groups treated with MOG: MOG9 vs. MOG21 (###, p < 0.001), MOG9 vs. MOG14 ( b , p < 0.05), MOG21 vs. MOG28 ( $$$ , p < 0.001), MOG14 vs. MOG21 ( aaa , p < 0.001). Bars show mean ± SEM; * p < 0.05, ** p < 0.01 and *** p < 0.001 vs. CFA using one-way ANOVA and Newman–Keuls post-test. Scale bars = 50 μm; (insets) = 20 μm.

Article Snippet: TRPV2 , Rabbit , 1:200 , ACC-032, Alomone , Biotinylated horse anti-rabbit , 1:500 , BA-1100, Vector Laboratories.

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: International Journal of Molecular Sciences

Article Title: TRPV2: A Key Player in Myelination Disorders of the Central Nervous System

doi: 10.3390/ijms23073617

Figure Lengend Snippet: MSRA and TRPV2 expression in frontal cortex samples from healthy subjects ( n = 4) and MS ( n = 6) patients. ( A ) TRPV2 mRNA ( p = 0.3314) is not changed between MS and healthy samples. ( B ) A significant upregulation in MSRA mRNA ( p = 0.0014) is found in MS; ( C ) A significant decrease in TRPV2 protein levels ( p = 0.0232) and a significant increase in MSRA protein levels ( p = 0.0124) were determined in MS samples by western blot. Bars show mean ± SEM; * p < 0.05, ** p < 0.01 using unpaired Student’s t -test.

Article Snippet: TRPV2 , Rabbit , 1:200 , ACC-032, Alomone , Biotinylated horse anti-rabbit , 1:500 , BA-1100, Vector Laboratories.

Techniques: Expressing, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: TRPV2: A Key Player in Myelination Disorders of the Central Nervous System

doi: 10.3390/ijms23073617

Figure Lengend Snippet: List of antibodies and reagents used in the slot blot, immunocytochemistry (ICC), immunohistochemistry (IHC), double immunofluorescence (IF) and western blot (WB).

Article Snippet: TRPV2 , Rabbit , 1:200 , ACC-032, Alomone , Biotinylated horse anti-rabbit , 1:500 , BA-1100, Vector Laboratories.

Techniques: Dot Blot, Immunocytochemistry, Immunohistochemistry, Immunofluorescence, Western Blot, Plasmid Preparation

Journal: International Journal of Molecular Sciences

Article Title: TRPV2: A Key Player in Myelination Disorders of the Central Nervous System

doi: 10.3390/ijms23073617

Figure Lengend Snippet: Primers used for quantitative polymerase chain reaction (qPCR).

Article Snippet: TRPV2 , Rabbit , 1:200 , ACC-032, Alomone , Biotinylated horse anti-rabbit , 1:500 , BA-1100, Vector Laboratories.

Techniques: Real-time Polymerase Chain Reaction

Journal: International Journal of Molecular Sciences

Article Title: TRPV2: A Key Player in Myelination Disorders of the Central Nervous System

doi: 10.3390/ijms23073617

Figure Lengend Snippet: TRPV2-centered cellular cross-talk model in physiological and pathophysiological myelination. During embryonic development, TRPV2 develops an essential role in neurite and axon outgrowth. In that period, TRPV2 is known to be expressed and activated in neurons, and putatively in OPCs, where possible cross-talk between both cells might be triggered through TRPV2 activation. In adulthood, TRPV2 expression is found in microglia, neurons and oligodendrocytes. Upon an insult such as demyelination, TRPV2, as a non-selective ion channel, is activated. Some known triggering stimuli are NO, LPC or oxidative stress, the latter causing methionine oxidation which facilitates TRPV2 sensitization. TRPV2 activation promotes a sodium/calcium inward influx from extracellular media and/or ER or endosomes. Consequently, cells are depolarized, which initiate processes such as actin polymerization or phagocytosis in microglia or innate immune cells. In those cells, TRPV2 has been involved in migration, phagocytosis or cytokine production. TRPV2 inactivation is promoted by MSRA enzyme, by reducing oxidized methionine. In our results, we observed both an increase of TRPV2 and MSRA in demyelinating scenarios. During remyelination, OPCs proliferate and migrate to the demyelinated area, where they differentiate through several stages to mature oligodendrocytes, and start myelination. Our results showed increased TRPV2 expression during this period. Overall, ubiquitous TRPV2 expression and activation in de- and remyelinating processes suggest a multicellular cross-talk that promotes myelin repair at all levels. TRPV2 is relevant in development and demyelination, making this model suitable for the recapitulation hypotheses. Created with BioRender.com (accessed on 10 January 2022) [ , , ].

Article Snippet: TRPV2 , Rabbit , 1:200 , ACC-032, Alomone , Biotinylated horse anti-rabbit , 1:500 , BA-1100, Vector Laboratories.

Techniques: Activation Assay, Expressing, Migration

Journal: Nature Communications

Article Title: Chromogranin A deficiency attenuates tauopathy by altering epinephrine–alpha-adrenergic receptor signaling in PS19 mice

doi: 10.1038/s41467-025-59682-6

Figure Lengend Snippet: A Heatmap representation of normalized read Z-scores of shared upregulated or downregulated genes in WT ( n = 5) and CgA-KO/PS19 ( n = 6) relative to PS19 ( n = 6) (log2 fold-change > 0.5, p value < 0.01) from 9-month-old mice cortices. Adra gene family showing significant expression changes are highlighted. B Z-score of hierarchical clusters I of WT, PS19 and CgA-KO/PS19. P-value was calculated using One-way Anova (F 2, 1248 = 163.9, P < 0.0001) Sidak’s multiple comparison test. C Z-score of hierarchical clusters II of WT, PS19 and CgA-KO/PS19. P-value was calculated using One-way Anova (F 2, 876 = 45.91, P < 0.0001) Sidak’s multiple comparison test. D Venn diagram showing number of upregulated genes as compared in two groups (WT/PS19 and CgA-KO/PS19). Expression of 417 genes overlaps in PS19 vs. WT and PS19 vs. CgA-KO/PS19. E Gene ontology analysis for pathways enriched in shared genes in ( D ). Number of genes involved in each pathway are shown in the bar. F Venn diagram showing number of downregulated genes as compared in two groups (WT/PS19 and CgA-KO/PS19). Expression of 293 genes overlaps in PS19 vs. WT and PS19 vs. CgA-KO/PS19. G Gene ontology analysis for pathways enriched in shared genes in ( E ) Number of genes involved in each pathway are shown in the bar. H Representative WB showing Adra1b, Adra2b and Adrb2 protein levels in three mouse groups, WT ( n = 4), PS19 ( n = 4) and CgA-KO/PS19 ( n = 4). Also shown are the levels of p-Tau (S202/T205), total PS19, CgA and tubulin (loading control). I Quantitation showing relative levels of Adra1b in ( H ). WT ( n = 4), PS19 ( n = 4) and CgA-KO/PS19 ( n = 4). P-value was calculated using One-way Anova (F 2, 9 = 0.07055, P = 0.0004) Sidak’s multiple comparison test. J Quantitation showing relative levels of Adra2b in ( H ). WT ( n = 4), PS19 ( n = 4) and CgA-KO/PS19 ( n = 4). P-value was calculated using One-way Anova (F 2, 9 = 0.8729, P = 0.0004) Sidak’s multiple comparison test. K Representative IF images of Adra1b, NeuN and DAPI in the hippocampus slices of PS19, CgA-KO/PS19 and WT mice. Scale bar = 200 μm. L Image ( J ) Quantitation of IF images as in ( H ) showing relative levels of Adra1b in PS19 ( n = 5) and CgA-KO/PS19 ( n = 5). P-value was calculated using (Unpaired two-tailed T-test with Welch’s correction, t = 2.649, df = 4.293). M Pearson correlation between Adra1b area fraction and the hippocampus volumes of PS19 (red dots) and CgA-KO/PS19 (blue dots) mice. Data are presented as mean values +/− SEM. Source data are provided as a Source Data file.

Article Snippet: Antibodies and dilution used as follows: AT8 (1:3000, Thermo Fisher Science Cat# MN1020), PHF1 (1:4000, Gift from P. Davies), CP13 (1:2000, Gift from P. Davies), HT7 (1:8000, Thermo Fisher Science Cat# MN1000), Actin (1:8000, Bio-Bharati, Cat# BB-AB0024), ADRA1B (1:2000, Abcam, Cat# AB 169523), ADRA2B (1:2000, Proteintech, Cat# 19778-1-AP), ADRB2 (1:2000, Cell Signaling Technology, Cat# 8513S), Alpha-Tubulin (1:6000, Cell Signaling Technology, Cat# 3873), CgA (1:2000, Invitrogen Cat# MA5-13093), Anti-Mouse-HRP (1:10000, Sigma, Cat# A0168), Anti-Rabbit-HRP (1:10000, Sigma, Cat# A0545), Strep-HRP (1:6000, Thermo Fisher Science Cat#N100).

Techniques: Expressing, Comparison, Control, Quantitation Assay, Two Tailed Test

Journal: Neurobiology of disease

Article Title: Inhibitory designer receptors aggravate memory loss in a mouse model of down syndrome

doi: 10.1016/j.nbd.2019.104616

Figure Lengend Snippet: α-ARs immunostaining in response to DREADD hM4Di inhibition of the LC. For all α-ARs, the main effects were attributable to treatment since compensatory receptor expressions were seen in (A) Regions examined by α-AR IF. Representative immunofluorescence for α1a-AR comparing all cohorts (parietal region) (B). Normalized fluorescent intensities for (C) α1a-AR (F1,56 = 9.45, p = .003), (D) α2a-AR (F1,56 = 17.2, p = .0001), (E) α2b-AR (F1,44 = 43.91, p < .0001) and (F) α2c-AR (F1,44 = 26.40, p < .0001). A significant effect could also be attributed to karyotype in α1a-AR (F1,56 = 8.40, p = .005), and α2c-AR (F1,44 = 17.73, p = .0001). Scale bar = 100 μm. (G) Expression levels of α2a-, α2b- and α2c-ARs negatively correlated with 24-h NORT DI performance. Error bars represent the mean ± SEM. Dotted lines represented the 95% confidence interval.

Article Snippet: For ARs, sections were incubated overnight at room temperature with anti-β1 (#AAR-023, dil. 1:400), anti-β2 (#AAR-016, dil. 1:200), anti-β3 (#AAR-017, dil. 1:100), anti-α1a (#AAR-015, dil. 1:100), anti-α2a (#AAR-020, dil. 1:200), anti-α2b (#AAR-021, dil. 1:100), or anti-α2c (#AAR-021, dil. 1:100) AR antibodies with or without blocking peptides at the recommended concentrations (Alomone Labs, Jerusalem, Israel).

Techniques: Immunostaining, Inhibition, Immunofluorescence, Expressing