Journal: PLoS ONE

Article Title: Chronic Ingestion of H1-Antihistamines Increase Progression of Atherosclerosis in Apolipoprotein E-/- Mice

doi: 10.1371/journal.pone.0102165

Figure Lengend Snippet: Atherosclerotic lesions around the aortic root of ApoE −/− mice were subjected to immunohistochemical analyses for H1R expression with anti-H1R (green) and DAPI (nuclear stain-blue). Panel (A) represents immunoflourescent staining for placebo (Pl), cetirizine low (C.L), cetirizine high (C.H), fexofenadine low (F.L) and fexofenadine high (F.H). Panel (B) depicts the integrated density of H1R staining. Magnification 40× at objective. n = 4.

Article Snippet: H1R protein expression in the atheroma was assessed in histological sections by using anti-H1R antibody (Alomone Labs, Israel) followed by Alexafluor-488 staining for immunofluorescence detection.

Techniques: Immunohistochemical staining, Expressing, Staining

Journal: bioRxiv

Article Title: THE CONSTITUTIVE ACTIVITY OF THE HISTAMINE H1 RECEPTOR INTERACTION WITH THE NMDA RECEPTOR : CONSEQUENCES IN EPILEPSY

doi: 10.1101/2021.11.02.466417

Figure Lengend Snippet: Immunostaining for HUC/ D (A), BMP (B) and GFAP (5C) co-expression with the H1 receptor in cultured hippocampal cells. Scale bar, 10 μm

Article Snippet: List of antibodies (dilution): Immunofluorescent labelling Anti-alpha 1 for GABAA receptor (1:1,000) (NeuroMab) anti-NMDA (1:1000) Ionotropic glutamate receptor subunit GluN1 (Synaptic Systems-SYSY) anti-H1 (1:100) ((Alomone Labs - AL) anti-GFAP (1:750) ( GFAP, Glial fibrillary acidic protein of astrocyte, (Chemicon, Temecula, CA) anti-BMP, bone morphogenetic protein of oligodendrocyte, (1:250) (Chemicon, Temecula, CA) anti-neuronal (protéine HUC/D.somatostatin (1:30) (Invitrogen)

Techniques: Immunostaining, Expressing, Cell Culture

Journal: bioRxiv

Article Title: THE CONSTITUTIVE ACTIVITY OF THE HISTAMINE H1 RECEPTOR INTERACTION WITH THE NMDA RECEPTOR : CONSEQUENCES IN EPILEPSY

doi: 10.1101/2021.11.02.466417

Figure Lengend Snippet: Immunostaining fo r NMDA receptor (A) and receptor GABA A receptor (B) co-expression with the H1 receptor in cultured hippocampal cells. Scale bar 10 μm and 2 μm as indicated

Article Snippet: List of antibodies (dilution): Immunofluorescent labelling Anti-alpha 1 for GABAA receptor (1:1,000) (NeuroMab) anti-NMDA (1:1000) Ionotropic glutamate receptor subunit GluN1 (Synaptic Systems-SYSY) anti-H1 (1:100) ((Alomone Labs - AL) anti-GFAP (1:750) ( GFAP, Glial fibrillary acidic protein of astrocyte, (Chemicon, Temecula, CA) anti-BMP, bone morphogenetic protein of oligodendrocyte, (1:250) (Chemicon, Temecula, CA) anti-neuronal (protéine HUC/D.somatostatin (1:30) (Invitrogen)

Techniques: Immunostaining, Expressing, Cell Culture

Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

Article Title: Histamine-mediated autocrine signaling in mesenteric perilymphatic mast cells

doi: 10.1152/ajpregu.00255.2019

Figure Lengend Snippet: Role of histamine receptors (HRs) in mast cells (MCs) in acute inflammation-induced mesenteric perilymphatic mast cell-mediated changes of tissue histamine. A: a significant proportion of mesenteric MCs are located near mesenteric lymphatic vessels [MLVs; A.1, bright-field, MLV identified by its intraluminal valve, MCs stained with avidin conjugate (A.2) and with toluidine blue (TB; A.3–A.4)]. Scale bars, 80 μm. B: illustration of surgical isolation of mesenteric perilymphatic tissue segments (B.2) from intestinal loops (B.1). B.3 shows a live single MLV from isolated segment presented in B.2. C: effects of various experimental treatments on histamine concentration in mesenteric perilymphatic tissues (untreated control, treatment with LPS; Crmln + LPS, pretreatment with cromolyn sodium before and during treatment with LPS; MC H1R active + LPS, pretreatment with HR2–4 antagonists before and during treatment with LPS; MC H2R active + LPS, pretreatment with HR1 and -3/4 antagonists before and during treatment with LPS; MC H3R + H4R + LPS, pretreatment with HR1 and -2 antagonists before and during treatment with LPS [n = 6 rats, normalized to untreated control; *significant differences (P < 0.05) between certain treatments]. D: illustration of web-based STRING platform analysis of predicted protein-protein interactions between subtypes of HRs and the histamine-producing enzyme histidine decarboxylase (HDC): H1R-HDC (E.1), H2R-HDC (E.2). E and F: effects of various experimental treatments on expression of HDC in mesenteric perilymphatic MCs (untreated control, 48/80, treatment with compound 48/80; LPS, treatment with LPS; Crmln + LPS, pretreatment with cromolyn sodium before and during treatment with LPS; H1B + LPS or H2B + LPS, pretreatment with HR1 antagonist or HR2 antagonist before and during treatment with LPS). E: mean fluorescence intensity (MFI) of HDC measured in mesenteric perilymphatic MCs after various experimental treatments [n = 4 rats, normalized to untreated control; *significant differences (P < 0.05) between certain treatments]. F: representative images of mesenteric segments containing MCs stained by Alexa fluor 488-conjugated avidin, in green), additionally labeled for HDC (in red), and DAPI (in blue) after various experimental treatments. Scale bars, 50 μm.

Article Snippet: Antibodies were obtained from the following sources: phospho-NF-κB p65 from Cell Signaling Technology, (cat. no. 3033S; Boston, MA), histidine decarboxylase (HDC) from Abcam (cat. no. ab37291; Boston, MA), HR1 (cat. no. AHR-001) and HR2 (cat. no. AHR-002) from Almone Laboratories, Jerusalem, Israel; HR2 from Novus Biologicals (cat. no. NLS1175), and CD11b/c from Biolegend (cat. no. 201801; San Diego, CA).

Techniques: Staining, Avidin-Biotin Assay, Isolation, Concentration Assay, Expressing, Fluorescence, Labeling

Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

Article Title: Histamine-mediated autocrine signaling in mesenteric perilymphatic mast cells

doi: 10.1152/ajpregu.00255.2019

Figure Lengend Snippet: Relative roles of histamine receptors (HRs) 1 and 2 in acute inflammation-induced mast cell (MC) degranulation. A: representative images of all variants of MC degranulation status. MCs stained with avidin or toluidine blue (T.Blue). B: quantitative analysis of variants of MC degranulation status under different experimental conditions (untreated control; LPS, treatment with LPS; pretreatment with HR1 antagonist before and during treatment with LPS (H1B + LPS); and pretreatment with HR2 antagonist before and during treatment with LPS (H2B + LPS); n = 4 rats, average mean values presented. C: representative images of MCs under each experimental condition stained with either avidin or T.Blue. Scale bars, 65 μm.

Article Snippet: Antibodies were obtained from the following sources: phospho-NF-κB p65 from Cell Signaling Technology, (cat. no. 3033S; Boston, MA), histidine decarboxylase (HDC) from Abcam (cat. no. ab37291; Boston, MA), HR1 (cat. no. AHR-001) and HR2 (cat. no. AHR-002) from Almone Laboratories, Jerusalem, Israel; HR2 from Novus Biologicals (cat. no. NLS1175), and CD11b/c from Biolegend (cat. no. 201801; San Diego, CA).

Techniques: Staining, Avidin-Biotin Assay

Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

Article Title: Histamine-mediated autocrine signaling in mesenteric perilymphatic mast cells

doi: 10.1152/ajpregu.00255.2019

Figure Lengend Snippet: Presence and functionality of histamine receptors (HRs) 1 and 2 in perilymphatic mast cells (MCs). A: representative confocal images of mesenteric perilymphatic tissue segments stained in red for HR1 (A.1) and HR2 (A.2) together with labeling of the same segment by Alexa fluor 488-conjugated avidin (in green) and DAPI (in blue). Scale bars, 20 μm. B: mean fluorescence intensity (MFI) of fluorescently labeled histamine measured in mesenteric perilymphatic MCs after various experimental treatments: H1B, pretreatment with H1R antagonist; H2B, pretreatment with H2R antagonist; H1H2B, pretreatment with both HR1 and HR2 antagonists [n = 3 rats, normalized to untreated control; *significant differences (P < 0.05) between treatment groups and control]. C: representative images of mesenteric segments containing MCs (stained by Texas Red-conjugated avidin, in red) bound with fluorescently labeled histamine (FLU-His, in green) after various experimental treatments. Scale bars, 50 μm.

Article Snippet: Antibodies were obtained from the following sources: phospho-NF-κB p65 from Cell Signaling Technology, (cat. no. 3033S; Boston, MA), histidine decarboxylase (HDC) from Abcam (cat. no. ab37291; Boston, MA), HR1 (cat. no. AHR-001) and HR2 (cat. no. AHR-002) from Almone Laboratories, Jerusalem, Israel; HR2 from Novus Biologicals (cat. no. NLS1175), and CD11b/c from Biolegend (cat. no. 201801; San Diego, CA).

Techniques: Staining, Labeling, Avidin-Biotin Assay, Fluorescence

Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

Article Title: Histamine-mediated autocrine signaling in mesenteric perilymphatic mast cells

doi: 10.1152/ajpregu.00255.2019

Figure Lengend Snippet: Histamine itself is able to induce activation of perilymphatic mast cells (MCs). A: representative images of MCs stained with both ruthenium red (R.Red) and toluidine blue (T.Blue) under different experimental conditions [untreated control (A.1 and A.2); 48/80 (treatment with compound 48/80, A.3 and A.4); treatment with histamine (A.5 and A.6); pretreatment by cromolyn before and during treatment with histamine (Crmln + Histamine, A.7 and A.8); pretreatment by histamine receptor 1 (HR1) antagonist before and during treatment with histamine (H1B + Histamine, A.9 and A.10); and pretreatment by HR2 antagonist before and during treatment with histamine (H2B + Histamine, A.11 and A.12)]. Insets from each picture demonstrate detailed structure of MCs under experimental conditions. Scale bars, 200 μm. B: quantitative analysis of mesenteric perilymphatic MC activation under various experimental conditions [n = 3 rats; *significant differences (P < 0.05) between corresponding experimental conditions]. C: schema of the MC-histamine autocrine regulatory loop. HDC, histidine decarboxylase; TLR4. Toll-like receptor 4.

Article Snippet: Antibodies were obtained from the following sources: phospho-NF-κB p65 from Cell Signaling Technology, (cat. no. 3033S; Boston, MA), histidine decarboxylase (HDC) from Abcam (cat. no. ab37291; Boston, MA), HR1 (cat. no. AHR-001) and HR2 (cat. no. AHR-002) from Almone Laboratories, Jerusalem, Israel; HR2 from Novus Biologicals (cat. no. NLS1175), and CD11b/c from Biolegend (cat. no. 201801; San Diego, CA).

Techniques: Activation Assay, Staining

Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

Article Title: Histamine-mediated autocrine signaling in mesenteric perilymphatic mast cells

doi: 10.1152/ajpregu.00255.2019

Figure Lengend Snippet: Mast cell (MC)-histamine autocrine regulatory loop: functional implications in mesenteric perilymphatic tissue compartments. A: expression of phosphorylated (p)NF-κB in mesenteric perilymphatic tissue segments under different experimental treatments [(untreated control; LPS-treated; Crmln + LPS, pretreatment with cromolyn sodium before and during treatment with LPS; H1B + LPS, pretreatment with histamine receptor 1 (HR1) antagonist before and during treatment with LPS; H2B + LPS, pretreatment with HR2 antagonist before and during treatment with LPS; H3/4B + LPS, pretreatment with HR3/4 antagonist before and during treatment with LPS (n = 4 rats, normalized to untreated control; *significant differences (P < 0.05) between certain treatments)]. Bottom: representative images of mesenteric perilymphatic tissue segments following various treatments. Scale bars, 80 μm. B: numerous cell types beyond MCs express HR1 and -2 in mesenteric perilymphatic tissues. Representative images showing immunohistochemical staining for HR1 or -2 (in red) and cell nuclei (DAPI, in blue) along with avidin conjugate to indicate MCs (in green). Scale bars, 50 μm. C: almost all CD11b/c-positive cells (in green, together with DAPI staining, in blue) express both HR1 and HR2 (in red): representative images. Scale bars, 20 μm. D and E: activation of MCs significantly increased the number of MCs that appeared physically associated with CD11b/c-positive cells. D: results of quantitative analysis [n = 4 rats; *significant differences (P < 0.05) between certain treatments: untreated control; 48/80, treatment with compound 48/80, LPS, treatment with LPS]. E: representative images (fluorescent labeling similar to 5B). Scale bars, 50 μm.

Article Snippet: Antibodies were obtained from the following sources: phospho-NF-κB p65 from Cell Signaling Technology, (cat. no. 3033S; Boston, MA), histidine decarboxylase (HDC) from Abcam (cat. no. ab37291; Boston, MA), HR1 (cat. no. AHR-001) and HR2 (cat. no. AHR-002) from Almone Laboratories, Jerusalem, Israel; HR2 from Novus Biologicals (cat. no. NLS1175), and CD11b/c from Biolegend (cat. no. 201801; San Diego, CA).

Techniques: Functional Assay, Expressing, Immunohistochemical staining, Staining, Avidin-Biotin Assay, Activation Assay, Labeling

Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

Article Title: Histamine-mediated autocrine signaling in mesenteric perilymphatic mast cells

doi: 10.1152/ajpregu.00255.2019

Figure Lengend Snippet: Roles of mast cell (MC) activation and histamine receptors (HRs) 1 and 2 in trafficking of CD11b/c-positive cells toward mesenteric lymphatic vessels (MLVs). A: representative images of trafficking of CD11b/c-positive cells (in green) toward MLVs under different experimental treatments [untreated control; 48/80, treatment with compound 48/80; LPS, treatment with LPS; Crmln + LPS, pretreatment with cromolyn sodium before and during treatment with LPS; H1B + LPS, pretreatment with HR1 antagonist before and during treatment with LPS; H2B + LPS, pretreatment with HR2 antagonist before and during treatment with LPS]. Scale bar, 100 μm. B: results of quantitative analysis [n = 4 rats; *significant differences (P < 0.05) between certain treatments]. C: schematic presentation of involvement of perilymphatic MCs in regulation of trafficking of CD11b/c-positive cells toward MLVs in response to LPS-induced acute inflammation. LEC, lymphatic endothelial cell; TLR4, Toll-like receptor 4; VCAM1, vascular cell adhesion molecule 1.

Article Snippet: Antibodies were obtained from the following sources: phospho-NF-κB p65 from Cell Signaling Technology, (cat. no. 3033S; Boston, MA), histidine decarboxylase (HDC) from Abcam (cat. no. ab37291; Boston, MA), HR1 (cat. no. AHR-001) and HR2 (cat. no. AHR-002) from Almone Laboratories, Jerusalem, Israel; HR2 from Novus Biologicals (cat. no. NLS1175), and CD11b/c from Biolegend (cat. no. 201801; San Diego, CA).

Techniques: Activation Assay

Journal: Journal of Cachexia, Sarcopenia and Muscle

Article Title: Histaminergic transmission slows progression of amyotrophic lateral sclerosis

doi: 10.1002/jcsm.12422

Figure Lengend Snippet: Histamine protects NSC‐34 G93A motor neurons via AKT/ERK1/2 pathways and rescues mitochondrial function. Representative confocal images of primary as well as differentiated NSC‐G93A motor neurons stained with H1‐H4R (green, A) or HDC, HNMT, DAO1 (green, C), and with SMI32 (red). Scale bar: 100 μm. Representative western blots and fold expression values of H1‐H4R (B) and HDC, HNMT, DAO1 (D) in NSC‐WT differentiated and ‐G93A differentiated cells. Values represent means from n = 3 independent experiments. Statistical significance was calculated by Student's t ‐test referred to NSC‐ WT cells, * P < 0.05. Representative western blots and quantification of pERK/ERK (E) and pAKT/AKT (F) in NSC‐WT and ‐G93A differentiated cells under serum starvation and in the absence or presence of histamine (HA, 10–1000 μM for 24 h). GAPDH was used as a loading control. Data represent means ± SEM. Statistical significance was calculated by analysis of variance (ANOVA) referred to NSC‐G93A untreated cells, * P < 0.05, n = 4 replicates. (G) Measurement of the rate of oxygen consumption ratio (OCR) in differentiated NSC‐WT and ‐G93A cell lines cultured without serum and treated with different concentrations of histamine (10–1000 μM for 24 h). Individual parameters for basal respiration, ATP production, maximal respiration, and spare respiratory capacity are indicated. Each data point represents an OCR measurement. Data represent means ± SEM. Statistical significance was calculated by ANOVA referred to untreated NSC‐G93A cells, * P < 0.05, n = 4 each performed in sextuplicate. Cell death in NSC‐G93A differentiated cells was determined under serum starvation in the absence or presence of histamine (10–1000 μM for 24 h) (H) or in cells exposed to histamine 1000 μM for 24 h and orphenadrine (H1R antagonist, 10 μM), ranitidine (H2R antagonist, 10 μM), thioperamide (H3R antagonist, 5 μM), PD98059 (500 nM), or wortmannin (10 nM) (I). Data represent means ± SEM. Statistical significance was calculated by ANOVA referred to cells in serum starvation medium, * P < 0.05, n = 4 performed in triplicate. DAO1, diamine oxidase 1; HDC, histidine decarboxylase; HNMT, histamine N‐methyltransferase; SEM, standard error of the mean; SOD1, superoxide dismutase 1; WT, wild‐type.

Article Snippet: Antibodies raised in rabbit: anti‐arginase 1 (ARG1) (1:100, Abcam, UK); anti‐CD163 (1:100, Santa Cruz Biotechnology, USA); anti‐choline acetyltransferase (1:500, AbCam); anti‐DAO1 (1:200, Bioss, USA); anti‐H1R (1:200, Alomone, Israel); anti‐H2R (1:200, MyBioSource, USA); anti‐H3R (1:200, Alomone); anti‐H4R (1:200, Santa Cruz Biotechnology); anti‐HDC (1:20, AbCam); anti‐HNMT (1:200, Sigma); anti‐inducible nitric oxide synthase (iNOS) (1:1000, CST, USA); anti‐myelin basic protein (MBP) (1:1000 CST); anti‐nuclear factor‐kappa B (NF‐κB) p65 (1:500, CST); anti‐phospho‐AKT (1:500, CST); anti‐phospho‐p44/42 MAPK (ERK1/2) (Thr202/Tyr204) (1:1000, CST); and anti‐phospho‐NF‐κB p65 (Ser536) (1:500, CST).

Techniques: Staining, Western Blot, Expressing, Cell Culture