Journal: Nature Metabolism

Article Title: A food-sensitive olfactory circuit drives anticipatory satiety

doi: 10.1038/s42255-025-01301-1

Figure Lengend Snippet: a , Paradigm of food odour exposure (NCD) and subsequent phosphorylated ribosome pulldown of the SR in fasted C57BL/6N mice. b , Quantification comparison of food odour-induced mRNA expression of neurons expressing Calb2 , Nts , Slc17a6 , Slc32a1 and Sst as the fold change between pulldown and input sample (two-way analysis of variance (ANOVA); n = 34 pooled groups of five mice each; P = 0.0001). c , Representative images of Slc17a6 (Vglut2) expression in the SR. Scale bars, 100 µm. d , Schematic of AAV-based expression of the calcium sensor GCaMP6s in MS VGLUT2 neurons and representative images of the fibre placement in VGLUT2-Cre mice (GCaMP6s, green; DAPI, blue). Scale bars, 100 µm. e , Paradigm for recording the activity dynamics of MS VGLUT2 neurons during food perception and food ingestion in freely behaving mice. f , MS VGLUT2 calcium dynamics and area under the curve (AUC) aligned to the onset of food consumption (NCD) in overnight-fasted mice; statistics indicate a significant difference between calcium signal before and after first bite (time curve: two-way ANOVA; n = 7; AUC: paired two-tailed Student’s t -test; n = 7 mice; P = 0.0068). g , MS VGLUT2 calcium dynamics before food ingestion ( n = 7 mice). h , Calcium dynamics of MS VGLUT2 neurons and AUC aligned to the first olfactory exploration (that is, sniff) when perceiving food (NCD) or a non-edible familiar object (empty teaball; control) in overnight-fasted mice; line on top indicates time points with a significant difference in calcium signal between control and NCD conditions (time curve: paired two-way ANOVA, Dunnett’s post hoc; n = 6 mice for control, n = 7 mice for NCD perception; AUC: unpaired two-tailed Student’s t -test; n = 6 mice for control, n = 7 mice for NCD perception; P = 0.024). Data are represented as the mean ± s.e.m. * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001, **** P ≤ 0.0001. Mouse images in d – f and Eppendorf image in a adapted from SciDraw under a Creative Commons license CC BY 4.0 . Panels a and h created using BioRender.com .

Article Snippet: The following inventoried TaqMan probes were used: Calb2 (Mm00801461_m1; Thermo Fisher Scientific), Nts (Mm00481140_m1; Thermo Fisher Scientific), Slc17a6 (Mm00499876_m1; Thermo Fisher Scientific), Slc32a1 (Mm00494138_m1; Thermo Fisher Scientific) and Sst (Mm00436671_m1; Thermo Fisher Scientific).

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

Journal: Brain research

Article Title: Detailed neuronal distribution of GPR3 and its co-expression with EF-hand calcium-binding proteins in the mouse central nervous system.

doi: 10.1016/j.brainres.2020.147166

Figure Lengend Snippet: Fig. 2. Distribution and characterization of GPR3-positive neurons in the ol- factory bulb using double fluorescent labeling in GPR3 knockout/LacZ knock-in mice. Representative images of the olfactory bulb stained with SPiDER-βGal (A) and Nissl (B) in heterozygous GPR3 knockout/LacZ knock-in mice are shown. Double fluorescent labeling of GPR3 knockout/LacZ knock-in mice was per- formed using antibodies for EF-hand Ca2+-binding proteins and SPiDER-βGal (for detailed methods see Experimental procedures). Representative images from the double staining of GPR3 with calbindin (C), CCK (D), calretinin (E), and TH (F) are shown. The percentage of GPR3-expressing cells that also ex- pressed each marker in the olfactory bulb was calculated (G). Data represent the % expression of each marker out of the total number of GPR3-expressing neurons in the olfactory bulb. The actual number of cells in each region is also shown (double-positive cells/total number of GPR3-positive cells). Data are expressed as the mean ± SE (n = 3 mice/group). Scale bars = 100 µm.

Article Snippet: For immunohistochemistry, at least two sections obtained from each mouse were incubated overnight at 4 °C in the following primary antibodies, all diluted at 1:400 in PBS: anti-parvalbumin mouse monoclonal antibody (MAB1572, Merck, Darmstadt, Germany), anti-calretinin rabbit polyclonal antibody (HPA007305, Atlas Antibody, Bromma, Sweden), anti-calbindin mouse monoclonal antibody (C9848, Sigma Aldrich, St. Louis, MO), anti-GAD67 mouse monoclonal antibody (MAB5406, Merck), anti-NECAB2 rabbit polyclonal antibody (HPA013998, Atlas Antibody), anti-tyrosine hydroxylase rabbit polyclonal antibody (AB152, Sigma Aldrich), anti-DARPP32 goat polyclonal antibody (AF6259, R&D Systems, Minneapolis, MN), anti-somatostatin rabbit polyclonal antibody (HPA019472, Atlas Antibody), anti-β3-tubulin rabbit monoclonal antibody (D71G9, Cell Signaling Technology, Danvers, MA), anti-CTIP2 rat monoclonal antibody (25B6, Abcam, Cambridge, UK), anti-cholecystokinin rabbit polyclonal antibody (C2581, Sigma Aldrich), and anti-NeuN mouse monoclonal antibody (MAB377, Sigma Aldrich).

Techniques: Labeling, Knock-Out, Knock-In, Staining, Binding Assay, Double Staining, Expressing, Marker

Journal: Brain research

Article Title: Detailed neuronal distribution of GPR3 and its co-expression with EF-hand calcium-binding proteins in the mouse central nervous system.

doi: 10.1016/j.brainres.2020.147166

Figure Lengend Snippet: Fig. 5. Distribution and characterization of GPR3-positive neurons in the tha- lamus using double fluorescent labeling in GPR3 knockout/LacZ knock-in mice. Representative images of the thalamus stained with SPiDER-βGal (A) and Nissl (B) in heterozygous GPR3 knockout/LacZ knock-in mice are shown. Double fluorescent labeling of GPR3 knockout/LacZ knock-in mice was performed using antibodies for EF-hand Ca2+-binding proteins and SPiDER-βGal. Representative images from the thalamus with double staining of GPR3 with calretinin (C) and parvalbumin (D, E) are shown. The percentage of GPR3-ex- pressing cells that also expressed parvalbumin in the thalamus was calculated (F). Data represent the % expression of each marker out of the total number of GPR3-expressing neurons in the thalamus. The actual number of cells in each region is also shown (double-positive cells/total number of GPR3-positive cells). Data are expressed as the mean ± SE (n = 3 mice/group). Scale bars = 500 µm. (caption on next page)

Article Snippet: For immunohistochemistry, at least two sections obtained from each mouse were incubated overnight at 4 °C in the following primary antibodies, all diluted at 1:400 in PBS: anti-parvalbumin mouse monoclonal antibody (MAB1572, Merck, Darmstadt, Germany), anti-calretinin rabbit polyclonal antibody (HPA007305, Atlas Antibody, Bromma, Sweden), anti-calbindin mouse monoclonal antibody (C9848, Sigma Aldrich, St. Louis, MO), anti-GAD67 mouse monoclonal antibody (MAB5406, Merck), anti-NECAB2 rabbit polyclonal antibody (HPA013998, Atlas Antibody), anti-tyrosine hydroxylase rabbit polyclonal antibody (AB152, Sigma Aldrich), anti-DARPP32 goat polyclonal antibody (AF6259, R&D Systems, Minneapolis, MN), anti-somatostatin rabbit polyclonal antibody (HPA019472, Atlas Antibody), anti-β3-tubulin rabbit monoclonal antibody (D71G9, Cell Signaling Technology, Danvers, MA), anti-CTIP2 rat monoclonal antibody (25B6, Abcam, Cambridge, UK), anti-cholecystokinin rabbit polyclonal antibody (C2581, Sigma Aldrich), and anti-NeuN mouse monoclonal antibody (MAB377, Sigma Aldrich).

Techniques: Labeling, Knock-Out, Knock-In, Staining, Binding Assay, Double Staining, Expressing, Marker

Journal: Brain research

Article Title: Detailed neuronal distribution of GPR3 and its co-expression with EF-hand calcium-binding proteins in the mouse central nervous system.

doi: 10.1016/j.brainres.2020.147166

Figure Lengend Snippet: Fig. 7. Distribution and characterization of GPR3-positive neurons in the substantia nigra using double fluorescent labeling in GPR3 knockout/LacZ knock-in mice. Representative images of the substantia nigra stained with SPiDER-βGal (A) and Nissl (B) in heterozygous GPR3 knockout/LacZ knock-in mice. Double fluorescent labeling of GPR3 knockout/LacZ knock-in mice was performed using antibodies for EF-hand Ca2+-binding proteins and SPiDER-βGal. Representative images from double fluorescent labeling of GPR3 with TH (C), parvalbumin (D), calretinin (E), and calbindin (F) in the SNpr and TH (G) in the SNc are shown. Representative images from double fluorescent labeling of GPR3 with TH (H, I) in the VTA are shown. The fluorescent double-positive neurons are indicated by arrowheads. The percentage of GPR3-expressing cells that also expressed each marker in the SNpr was calculated (J). Data represent the % expression of each marker out of the total number of GPR3-expressing neurons in the SNpr. The actual number of cells in each region is also shown (double-positive cells/total number of GPR3-positive cells). Data are expressed as the mean ± SE (n = 3 mice/group). Scale bars = 500 µm (A, B, G, H), 100 µm (C–F).

Article Snippet: For immunohistochemistry, at least two sections obtained from each mouse were incubated overnight at 4 °C in the following primary antibodies, all diluted at 1:400 in PBS: anti-parvalbumin mouse monoclonal antibody (MAB1572, Merck, Darmstadt, Germany), anti-calretinin rabbit polyclonal antibody (HPA007305, Atlas Antibody, Bromma, Sweden), anti-calbindin mouse monoclonal antibody (C9848, Sigma Aldrich, St. Louis, MO), anti-GAD67 mouse monoclonal antibody (MAB5406, Merck), anti-NECAB2 rabbit polyclonal antibody (HPA013998, Atlas Antibody), anti-tyrosine hydroxylase rabbit polyclonal antibody (AB152, Sigma Aldrich), anti-DARPP32 goat polyclonal antibody (AF6259, R&D Systems, Minneapolis, MN), anti-somatostatin rabbit polyclonal antibody (HPA019472, Atlas Antibody), anti-β3-tubulin rabbit monoclonal antibody (D71G9, Cell Signaling Technology, Danvers, MA), anti-CTIP2 rat monoclonal antibody (25B6, Abcam, Cambridge, UK), anti-cholecystokinin rabbit polyclonal antibody (C2581, Sigma Aldrich), and anti-NeuN mouse monoclonal antibody (MAB377, Sigma Aldrich).

Techniques: Labeling, Knock-Out, Knock-In, Staining, Binding Assay, Expressing, Marker

Journal: Brain research

Article Title: Detailed neuronal distribution of GPR3 and its co-expression with EF-hand calcium-binding proteins in the mouse central nervous system.

doi: 10.1016/j.brainres.2020.147166

Figure Lengend Snippet: Fig. 9. Distribution and characterization of GPR3-positive neurons in the cer- ebellum using double fluorescent labeling in GPR3 knockout/LacZ knock-in mice. Representative images of the cerebellum stained with SPiDER-βGal (A) and Nissl (B) in heterozygous GPR3 knockout/LacZ knock-in mice. Double fluorescent labeling of GPR3 knockout/LacZ knock-in mice was performed using antibodies for EF-hand Ca2+-binding proteins and SPiDER-βGal. Representative images from double fluorescent labeling of GPR3 with calretinin (C) in the CGL of the cerebellum, and with calretinin (D) and parvalbumin (E) in the DCN are shown. The fluorescent double-positive neurons are indicated by arrowheads. The percentage of GPR3-expressing cells that also expressed each marker in the DCN of the cerebellum was calculated (F). Data represent the % expression of each marker out of the total number of GPR3-expressing neurons in the DCN of the cerebellum. The actual number of cells in each region is also shown (double-positive cells/total number of GPR3-positive cells). Data are expressed as the mean ± SE (n = 3 mice/group). Scale bars = 500 µm (A, B), 100 µm (C–E).

Article Snippet: For immunohistochemistry, at least two sections obtained from each mouse were incubated overnight at 4 °C in the following primary antibodies, all diluted at 1:400 in PBS: anti-parvalbumin mouse monoclonal antibody (MAB1572, Merck, Darmstadt, Germany), anti-calretinin rabbit polyclonal antibody (HPA007305, Atlas Antibody, Bromma, Sweden), anti-calbindin mouse monoclonal antibody (C9848, Sigma Aldrich, St. Louis, MO), anti-GAD67 mouse monoclonal antibody (MAB5406, Merck), anti-NECAB2 rabbit polyclonal antibody (HPA013998, Atlas Antibody), anti-tyrosine hydroxylase rabbit polyclonal antibody (AB152, Sigma Aldrich), anti-DARPP32 goat polyclonal antibody (AF6259, R&D Systems, Minneapolis, MN), anti-somatostatin rabbit polyclonal antibody (HPA019472, Atlas Antibody), anti-β3-tubulin rabbit monoclonal antibody (D71G9, Cell Signaling Technology, Danvers, MA), anti-CTIP2 rat monoclonal antibody (25B6, Abcam, Cambridge, UK), anti-cholecystokinin rabbit polyclonal antibody (C2581, Sigma Aldrich), and anti-NeuN mouse monoclonal antibody (MAB377, Sigma Aldrich).

Techniques: Labeling, Knock-Out, Knock-In, Staining, Binding Assay, Expressing, Marker

Journal: Brain research

Article Title: Detailed neuronal distribution of GPR3 and its co-expression with EF-hand calcium-binding proteins in the mouse central nervous system.

doi: 10.1016/j.brainres.2020.147166

Figure Lengend Snippet: Fig. 10. Distribution and characterization of GPR3-positive neurons in the spinal cord using double fluorescent labeling in GPR3 knockout/LacZ knock-in mice. Representative images of the spinal cord stained with SPiDER-βGal (A) and Nissl (B) in heterozygous GPR3 knockout/LacZ knock-in mice. Double fluorescent labeling of GPR3 knockout/LacZ knock-in mice was performed using antibodies for EF-hand Ca2+-binding proteins and SPiDER-βGal. Representative images from double fluorescent labeling of GPR3 with calretinin (C) and NECAB2 (D) in the AH of the spinal cord are shown. The fluorescent double-positive neurons are indicated by arrowheads. The percentage of GPR3- expressing cells that also expressed calretinin in the spinal cord was calculated (E). Data represent the % expression of calretinin out of the total number of GPR3-expressing neurons in the spinal cord. The actual number of cells in each region is also shown (double-positive cells/total number of GPR3-positive cells). Data are expressed as the mean ± SE (n = 3 mice/group). Scale bars = 500 µm (A, B), 100 µm (C, D).

Article Snippet: For immunohistochemistry, at least two sections obtained from each mouse were incubated overnight at 4 °C in the following primary antibodies, all diluted at 1:400 in PBS: anti-parvalbumin mouse monoclonal antibody (MAB1572, Merck, Darmstadt, Germany), anti-calretinin rabbit polyclonal antibody (HPA007305, Atlas Antibody, Bromma, Sweden), anti-calbindin mouse monoclonal antibody (C9848, Sigma Aldrich, St. Louis, MO), anti-GAD67 mouse monoclonal antibody (MAB5406, Merck), anti-NECAB2 rabbit polyclonal antibody (HPA013998, Atlas Antibody), anti-tyrosine hydroxylase rabbit polyclonal antibody (AB152, Sigma Aldrich), anti-DARPP32 goat polyclonal antibody (AF6259, R&D Systems, Minneapolis, MN), anti-somatostatin rabbit polyclonal antibody (HPA019472, Atlas Antibody), anti-β3-tubulin rabbit monoclonal antibody (D71G9, Cell Signaling Technology, Danvers, MA), anti-CTIP2 rat monoclonal antibody (25B6, Abcam, Cambridge, UK), anti-cholecystokinin rabbit polyclonal antibody (C2581, Sigma Aldrich), and anti-NeuN mouse monoclonal antibody (MAB377, Sigma Aldrich).

Techniques: Labeling, Knock-Out, Knock-In, Staining, Binding Assay, Expressing

Journal: Cells

Article Title: Combinatorial Genomic Biomarkers Associated with High Response in IgE-Dependent Degranulation in Human Mast Cells

doi: 10.3390/cells13151237

Figure Lengend Snippet: Relative expression levels of the 8 genes selected for TaqMan PCR analysis of samples from High, Average and Low responders. Statistical analysis revealed that significant differences in expression levels of DPP4, GADD45B, NELL2, IL13RA1, AKAP12, CALB2 and ITM2C were detected between High vs. Low responders and High vs. Average responders. Data calculated from the statistical analysis of the comparisons between different groups for each of the 8 genes are shown in the table beneath the figures.

Article Snippet: TaqMan probes used are listed as the following: AKAP12 (Hs00374507_m1), ARHGAP15 (Hs00251227_m1), CALB2 (Hs00242372_m1), CEBPD (Hs00270931_s1), DPP4 (Hs00897386_m1), FCGR1A/B/CP (Hs02340030_m1), FOXF1 (Hs00230962_m1), GADD45B (Hs00169587_m1), GAPDH (Hs03929097_g1), IL13AR1 (Hs00609812_m1), IL1R1 (Hs00991010_m1), IRF1 (Hs00971965_m1), ITM2C (Hs00985194_g1), KCNMA1 (Hs01119504_m1), NELL2 (Hs00196254-m1), PTGDR (Hs00235003_m1), SIGLEC8 (Hs00274289_m1), SOCS2 (Hs00919620_m1), SPTLC3 (Hs00217867_m1), THEMIS (Hs01041269_m1), TMEM255B (Hs00415678_m1).

Techniques: Expressing

Journal: Cells

Article Title: Combinatorial Genomic Biomarkers Associated with High Response in IgE-Dependent Degranulation in Human Mast Cells

doi: 10.3390/cells13151237

Figure Lengend Snippet: PEEP profiling revealed the heterogeneity of expression levels of each of the 8 selected genes in different responder groups. For AKAP12, CALB2, ITM2C, IL13RA1, NELL2 and TMEM255B genes, whose expression levels were upregulated in the High responders as compared to those in other groups, their “% perturbed” and “% up” values in the High responders were higher than those calculated for other responder groups. For GADD45B and DPP4 genes, whose expression levels were downregulated in the High responders as compared to those in other groups, their “% perturbed” and “% up” values in the High responders were lower than those calculated for other responder groups.

Article Snippet: TaqMan probes used are listed as the following: AKAP12 (Hs00374507_m1), ARHGAP15 (Hs00251227_m1), CALB2 (Hs00242372_m1), CEBPD (Hs00270931_s1), DPP4 (Hs00897386_m1), FCGR1A/B/CP (Hs02340030_m1), FOXF1 (Hs00230962_m1), GADD45B (Hs00169587_m1), GAPDH (Hs03929097_g1), IL13AR1 (Hs00609812_m1), IL1R1 (Hs00991010_m1), IRF1 (Hs00971965_m1), ITM2C (Hs00985194_g1), KCNMA1 (Hs01119504_m1), NELL2 (Hs00196254-m1), PTGDR (Hs00235003_m1), SIGLEC8 (Hs00274289_m1), SOCS2 (Hs00919620_m1), SPTLC3 (Hs00217867_m1), THEMIS (Hs01041269_m1), TMEM255B (Hs00415678_m1).

Techniques: Expressing, Standard Deviation