Journal: Journal of Neuroinflammation

Article Title: Age-dependent involvement of gut mast cells and histamine in post-stroke inflammation

doi: 10.1186/s12974-020-01833-1

Figure Lengend Snippet: Elevated levels of histamine following ischemic stroke in a blood plasma of young (Yg) and aged (Ag) mice at 6 and 24-h post-stroke compared to age matched controls. b Histamine levels of brain quantified by mass spectrometry in aged (Ag) mice at 6 and 24-h post-stroke compared to age-matched sham control mice. n = 3 (sham) and 5 (stroke) per group. Data are expressed as mean ± SEM ( a ) and median ± range ( b ), as well as individual values. * P < 0.05, ** P < 0.01, *** P < 0.001. P values were calculated using one-way analysis of variance with Tukey multiple comparisons correction ( a ) and Kruskal-Wallis test with Dunn’s multiple comparison ( b )

Article Snippet: Formalin fixed, paraffin-embedded intestinal (cecal, ileum) tissue sections (4 μm) were incubated overnight at 4 °C with a primary antibody targeting the mouse antigens (1) histamine receptor 2 (AHR-002, Alomone labs, Israel), (2) Tryptase (ab2378, Abcam, USA), after antigen retrieval according to the manufacturer’s instructions.

Techniques: Clinical Proteomics, Mass Spectrometry, Control, Analysis, Comparison

Journal: Journal of Neuroinflammation

Article Title: Age-dependent involvement of gut mast cells and histamine in post-stroke inflammation

doi: 10.1186/s12974-020-01833-1

Figure Lengend Snippet: Increased levels of histamine receptor 2 (HR2) measured in the intestinal samples of young and aged mice for 6-h, 24-h, and 7-days post-stroke compared to the age matched controls. a mRNA expression levels of HR2 quantified by qPCR in young and aged mice at 6 and 24 h post-stroke. b (i) Protein levels of HR2 quantified by IHC followed by image J quantification in aged mice at 7 days post-stroke and c mRNA levels of HR2 quantified by RNA in situ hybridization and analyzed by image-J in the intestine of Ag mice at 7 days post-stroke. n = 3 (sham) and 5 (stroke) per group ( a ) and 5 (sham) and 5 (stroke) per group ( b and c ). Data are expressed as mean ± SEM ( a ), ( b ), and ( c ), as well as individual values. * P < 0.05, ** P < 0.01, *** P < 0.001. P values were calculated using one-way analysis of variance with Tukey multiple comparisons correction ( a ) or with two-tailed unpaired Student’s t test ( b and c ). b Brown-HR2 protein; blue-Nuclei ( b ) purple dots (red arrows)-mRNA HR2

Article Snippet: Formalin fixed, paraffin-embedded intestinal (cecal, ileum) tissue sections (4 μm) were incubated overnight at 4 °C with a primary antibody targeting the mouse antigens (1) histamine receptor 2 (AHR-002, Alomone labs, Israel), (2) Tryptase (ab2378, Abcam, USA), after antigen retrieval according to the manufacturer’s instructions.

Techniques: Expressing, RNA In Situ Hybridization, Analysis, Two Tailed Test, Immunohistochemistry

Journal: PLoS ONE

Article Title: Estrogenic Regulation of Histamine Receptor Subtype H1 Expression in the Ventromedial Nucleus of the Hypothalamus in Female Rats

doi: 10.1371/journal.pone.0096232

Figure Lengend Snippet: Quantitative analysis of H1R mRNA and H1R protein in the two subregions of the VMN was performed after selective dissection of the vl VMN and dm VMN using a stainless steel needle. The dotted lines show the locations of the VMN and vl and dm subregions in the rat basal hypothalamus. (A) Representative photograph of a coronal brain section after sampling (left side). The vl VMN and dm VMN were selectively dissected with the needle (inner diameter 900 µm). An outline of the rat basal hypothalamus used for identification and selective dissection of the VMN subregions. 3V, Third ventricle; VMN, ventromedial nucleus of the hypothalamus; vl, ventrolateral part of the VMN; dm, dorsomedial part of the VMN; Me, Median eminence; ARC, arcuate nucleus. Scale bars, 500 µm.

Article Snippet: The blotted membranes were blocked with 1.5% skim milk TBST at 4°C for 20 min and then incubated with rabbit polyclonal H1R antibodies (AHR-001; Alomone Labs, Jerusalem, Israel; 1∶1,000 dilution) and mouse monoclonal antibody to tubulin (T5201; Sigma Japan, Tokyo, Japan;1∶30,000 dilution in TBST) at 4°C for 12 h. The specificity of the AHR-001 antibody was shown in a preabsorption test with control peptide antigen (see product information for AHR-001; Alomone Labs, Jerusalem, Israel).

Techniques: Dissection, Sampling

Journal: PLoS ONE

Article Title: Estrogenic Regulation of Histamine Receptor Subtype H1 Expression in the Ventromedial Nucleus of the Hypothalamus in Female Rats

doi: 10.1371/journal.pone.0096232

Figure Lengend Snippet: (A–F) Confocal microscopy images of dual immunofluorescence labeling of ERα (green, A and D) and H1R (red, B and E) in adult female rats. ERα-ir neurons are abundant in the vl VMN, but few are present in the dm VMN (A). (B) The highest density of H1R expression was found in the vl VMN. Merged images showed the coexistence of abundant ERα-ir and H1R-ir in the VMN (C and F). (D–F) Medium magnification images of representative neural groups within the vl VMN (area with a dotted line), allowing identification of cellular localization of ERα and H1R. (G and H) Detection of H1R immunoreactivities in VMN with (H) or without (G) control peptide. Preabsorption of H1R antibody with its peptide eliminated H1R immunoreactivities (H). 3V, Third ventricle; VMN, ventromedial nucleus of the hypothalamus; vl, ventrolateral part of the VMN; dm, dorsomedial part of the VMN; Me, median eminence; ARC, arcuate nucleus. Scale bars, 500 µm (A–C), 50 µm (D–H).

Article Snippet: The blotted membranes were blocked with 1.5% skim milk TBST at 4°C for 20 min and then incubated with rabbit polyclonal H1R antibodies (AHR-001; Alomone Labs, Jerusalem, Israel; 1∶1,000 dilution) and mouse monoclonal antibody to tubulin (T5201; Sigma Japan, Tokyo, Japan;1∶30,000 dilution in TBST) at 4°C for 12 h. The specificity of the AHR-001 antibody was shown in a preabsorption test with control peptide antigen (see product information for AHR-001; Alomone Labs, Jerusalem, Israel).

Techniques: Confocal Microscopy, Immunofluorescence, Labeling, Expressing, Control

Journal: PLoS ONE

Article Title: Estrogenic Regulation of Histamine Receptor Subtype H1 Expression in the Ventromedial Nucleus of the Hypothalamus in Female Rats

doi: 10.1371/journal.pone.0096232

Figure Lengend Snippet: (A, B, C, D, E) Representative images of dual immunofluorescence labeling of ERα (green) and H1R (red), obtained by confocal microscopy. Higher magnification images of individual cells in panel A are shown in B (area with a dotted line), and C, D and E (arrowheads). C–E are confocal images showing that ERα is exclusively localized in nuclei (green) and excluded from nucleoli (dark spot), while H1R is localized in the cell membrane (red). Panel E is a merge of the images from panels C and D. (F, G, H) Visualization of the cellular localization of ERα-ir in VMN neurons. ERα immunostaining (green) were observed in the nucleus defined by PI staining (red). Panel H is the merged image for panel F and G. ERα; ERα immunoreactivities, PI; propidium iodide. Scale bars, 5 µm (A–E), 10 µm (F–H).

Article Snippet: The blotted membranes were blocked with 1.5% skim milk TBST at 4°C for 20 min and then incubated with rabbit polyclonal H1R antibodies (AHR-001; Alomone Labs, Jerusalem, Israel; 1∶1,000 dilution) and mouse monoclonal antibody to tubulin (T5201; Sigma Japan, Tokyo, Japan;1∶30,000 dilution in TBST) at 4°C for 12 h. The specificity of the AHR-001 antibody was shown in a preabsorption test with control peptide antigen (see product information for AHR-001; Alomone Labs, Jerusalem, Israel).

Techniques: Immunofluorescence, Labeling, Confocal Microscopy, Membrane, Immunostaining, Staining

Journal: PLoS ONE

Article Title: Estrogenic Regulation of Histamine Receptor Subtype H1 Expression in the Ventromedial Nucleus of the Hypothalamus in Female Rats

doi: 10.1371/journal.pone.0096232

Figure Lengend Snippet: Normalized H1R mRNA expression levels were quantified for males (n = 5), OVX females (oil injected controls, n = 5), OVX females with EB (5 µg EB, 24 h or 48 h post-injection, n = 5 each), and OVX females with EB + progesterone (5 µg EB, followed by 500 µg progesterone, n = 5). In the vl VMN, H1R mRNA was significantly increased by EB in OVX females and decreased by progesterone. In the dm VMN, hormonal manipulation did not affect H1R mRNA expression. Data represents the absolute quantities of H1R mRNA compared with GAPDH mRNA amount. Data are presented as the mean ±SEM. **, P<0.01; *, P<0.05.

Article Snippet: The blotted membranes were blocked with 1.5% skim milk TBST at 4°C for 20 min and then incubated with rabbit polyclonal H1R antibodies (AHR-001; Alomone Labs, Jerusalem, Israel; 1∶1,000 dilution) and mouse monoclonal antibody to tubulin (T5201; Sigma Japan, Tokyo, Japan;1∶30,000 dilution in TBST) at 4°C for 12 h. The specificity of the AHR-001 antibody was shown in a preabsorption test with control peptide antigen (see product information for AHR-001; Alomone Labs, Jerusalem, Israel).

Techniques: Expressing, Injection

Journal: PLoS ONE

Article Title: Estrogenic Regulation of Histamine Receptor Subtype H1 Expression in the Ventromedial Nucleus of the Hypothalamus in Female Rats

doi: 10.1371/journal.pone.0096232

Figure Lengend Snippet: H1R protein expression levels were quantified for males (n = 5), OVX females (oil injected controls, n = 5), OVX females with EB (5 µg EB, 24 h or 48 h post-injection, n = 5 each), and OVX females with EB+ progesterone (5 µg EB, followed by 500 µg progesterone, n = 5). In the vl VMN, the H1R protein level tended to be increased by EB in OVX females (not statistically significant) and decreased by progesterone. In the dm VMN, hormonal manipulation did not affect H1R protein expression. Data are presented as the mean ±SEM. **, P<0.01. Data are expressed as the ratios of H1R protein to tubulin protein in the vl VMN and dm VMN regions. Since the immunoblotting were obtained independently for each region (from the respective immunoblotting membranes for the vl VMN and dm VMN), data were not to be compared between the two regions.

Article Snippet: The blotted membranes were blocked with 1.5% skim milk TBST at 4°C for 20 min and then incubated with rabbit polyclonal H1R antibodies (AHR-001; Alomone Labs, Jerusalem, Israel; 1∶1,000 dilution) and mouse monoclonal antibody to tubulin (T5201; Sigma Japan, Tokyo, Japan;1∶30,000 dilution in TBST) at 4°C for 12 h. The specificity of the AHR-001 antibody was shown in a preabsorption test with control peptide antigen (see product information for AHR-001; Alomone Labs, Jerusalem, Israel).

Techniques: Expressing, Injection, Western Blot

Journal: Cells

Article Title: Potential Role of Intracranial Mast Cells in Neuroinflammation and Neuropathology Associated with Food Allergy

doi: 10.3390/cells11040738

Figure Lengend Snippet: Primary antibodies used for western blotting.

Article Snippet: Histamine H3 receptor (H3R) , Rabbit , Polyclonal , AHR-003 , 1:500 , Alomone Labs.

Techniques: Western Blot