Journal: Journal of nutritional science and vitaminology

Article Title: Effect of Oleuropein on Anti-Obesity and Uncoupling Protein 1 Level in Brown Adipose Tissue in Mild Treadmill Walking Rats with Diet-Induced Obesity.

doi: 10.3177/jnsv.70.193

Figure Lengend Snippet: Fig. 4. Protein levels of transient receptor potential ankyrin subtype 1 (TRPA1) (A), transient receptor potential vanilloid subtype 1 (TRPV1) (B), and brain-derived neurotrophic factor (BDNF) (C) and the results of Western blot (D) in the brain, plasma BDNF level (E), and BDNF (F) and peroxisome proliferator-activated receptor-gamma coactivator 1-alpha (PGC1- ) protein level in the gastrocnemius muscle (G) of rats fed with a high-fat diet (HF) or HF supplemented with 0.08% oleuro- pein (HFO) combined with mild treadmill walking (MTW) for 28 d. Values are expressed as meansSE, n 6 or 7. Means labeled with different letters indicate a signifi cant difference (p0.05). The effects of oleuropein supplementation (O) and MTW (W) and the interaction between O and W (OW) were both signifi cant (p0.05).

Article Snippet: The primary antibodies were as follows: BDNF (15 kDa, ab108319, 1 : 1,000; Abcam), TRPV1 (100 kDa, NBP1-97417, 1 : 1,000; Novus Bio), TRPA1 (100 kDa, NB11040763, 1 : 1,000; Novus Bio), peroxisome proliferatoractivated receptor-gamma coactivator 1-alpha (PGC1; 100 kDa, ab106814, 1 : 1,000; Abcam), -actin (42 kDa, ab185058, 1 : 10,000; Abcam), and -actin (42 kDa, ab20272, 1 : 5,000; Abcam).

Techniques: Derivative Assay, Western Blot, Clinical Proteomics, Labeling

Journal: Investigative Ophthalmology & Visual Science

Article Title: The Impact of Aging and Sex on Corneal Nerve Density and Function

doi: 10.1167/iovs.67.3.3

Figure Lengend Snippet: Aging does not affect the gene expression of corneal sensory receptors in the TG. ( A – D ) Quantification of TRPV1 ( A ), TRPA1 ( B ), TRPM8 ( C ), and PIEZO2 ( D ) gene expression in the TG of male and female mice 8 and 52 weeks old ( n = 4–6 mice/group). Graphs represent mean values ± SEM; Statistical analysis was performed using unpaired Student's t -tests.

Article Snippet: TaqMan Gene Expression Assays (Applied Biosystems, Foster City, CA, USA) for Tac1 (Mm01166996_m1), Tacr1 (Mm00436892_m1), Calca (Mm00801463_g1), Ramp1 (Mm00489796_m1), Sox11 (Mm01281943_s1), Gap43 (Mm00500404_m1), Gdnf (Mm00599849_m1), Atf3 (Mm00476033_m1), Trpv1 (Mm01246300_m1), Trpm8 (Mm01299593_m1), Piezo2 (Mm01265861_m1), Trpa1 (Mm01227437_m1), Ngf (Mm00443039_m1), Bdnf (Mm04230607_s1), and glyceraldehyde 3-phosphate dehydrogenase (Mm99999915_g1) as the housekeeping gene were performed.

Techniques: Gene Expression

Journal: International Journal of Molecular Sciences

Article Title: Role of Major Endocannabinoid-Binding Receptors during Mouse Oocyte Maturation

doi: 10.3390/ijms20122866

Figure Lengend Snippet: Expression levels of endocannabinoids (eCBs)-binding receptors during mouse oocyte in vivo meiotic maturation. ( A ) Real-time PCR of Cnr1 , Cnr2 , Gpr55 , and Trpv1 . Data were reported as 2 −ΔΔ C t values calculated by Delta–Delta Ct (ΔΔ C t) method vs. germinal vesicle (GV) ( Cnr1 ) group posed equal to 1. Expression was normalized to Actb and values were reported as mean ± SEM of 4 independent replicates. * p < 0.05 vs. GV oocyte of the same experimental group. ( B ) Representative western blot and quantification of cannabinoid receptor type-1 and -2 (CB 1 R and CB 2 R), G-protein coupled receptor 55 (GPR55), and transient receptor potential vanilloid type 1 channel (TRPV1) protein contents. Data are expressed as mean ± SEM of each receptor content after normalization with α/β-tubulin, used as loading control. Experiments were repeated 3 times. * p < 0.05 vs. GV oocyte of the same experimental group; ** p < 0.05 vs. MI of the same experimental group. GV = germinal vesicle; MI = metaphase I; MII = metaphase II.

Article Snippet: Rabbit TRPV1 (cat. TA336871) antibody was obtained from OriGene Technologies, Inc. (Rockville, MD, USA).

Techniques: Expressing, Binding Assay, In Vivo, Real-time Polymerase Chain Reaction, Western Blot, Control

Journal: International Journal of Molecular Sciences

Article Title: Role of Major Endocannabinoid-Binding Receptors during Mouse Oocyte Maturation

doi: 10.3390/ijms20122866

Figure Lengend Snippet: ( A ) Localization of CB 1 R, CB 2 R, GPR55, and TRPV1 receptors in mouse oocytes collected at various stages of in vivo meiotic maturation. Receptors were labelled with Cy-3 (red), DNA was counterstained by DAPI (cyan). In the upper right-hand corner of GV CB 1 R, the strong DAPI staining is due to undetached cumulus cell nuclei. Each image was taken at the equatorial plan of the oocyte. Magnification: ×630. Each inset represents a magnified part of ooplasm. GV = germinal vesicle; MI = metaphase I; MII = metaphase II; NC = negative control. ( B ) Mean fluorescence of CB 1 R, CB 2 R, GPR55, and TRPV1 receptors in mouse oocytes collected at different stages of meiotic maturation. Values are expressed as arbitrary units (AU) and are reported as mean ± SEM of 6 oocytes from 3 independent experiments. * p < 0.05 vs. GV oocyte of the same experimental group; ** p < 0.05 vs. MI of the same experimental group.

Article Snippet: Rabbit TRPV1 (cat. TA336871) antibody was obtained from OriGene Technologies, Inc. (Rockville, MD, USA).

Techniques: In Vivo, Staining, Negative Control, Fluorescence

Journal: International Journal of Molecular Sciences

Article Title: Role of Major Endocannabinoid-Binding Receptors during Mouse Oocyte Maturation

doi: 10.3390/ijms20122866

Figure Lengend Snippet: Distribution of CB 1 R, CB 2 R, GPR55, and TRPV1 in the oolemma and CB 1 R localization at oolemma during in vivo GV–MI transition. ( A ) Only GV oocytes showed detectable signals. Qualitative data are expressed as presence (+) or absence (-) of fluorescence in 15 oocytes/sample. ( B ) CB 1 R distribution, analyzed at different times after human chorionic gonadotropin (hCG), changed from uniformly homogeneous (0–3 h) to dotted clusters (5 h), until complete disappearance at MI (8 h). Each image was taken at the equatorial plan of the oocyte. Cy-3: red, DAPI: cyan. Magnification: ×630.

Article Snippet: Rabbit TRPV1 (cat. TA336871) antibody was obtained from OriGene Technologies, Inc. (Rockville, MD, USA).

Techniques: In Vivo, Fluorescence

Journal: International Journal of Molecular Sciences

Article Title: Role of Major Endocannabinoid-Binding Receptors during Mouse Oocyte Maturation

doi: 10.3390/ijms20122866

Figure Lengend Snippet: List of the primers used for quantitative real-time PCR analysis.

Article Snippet: Rabbit TRPV1 (cat. TA336871) antibody was obtained from OriGene Technologies, Inc. (Rockville, MD, USA).

Techniques: Real-time Polymerase Chain Reaction

Journal: Journal of advanced research

Article Title: The LRP1-SHP2 pathway regulates TRPV1 sensitivity in the peripheral nervous system: Insights from amyloid beta 1-42 modulation.

doi: 10.1016/j.jare.2025.03.005

Figure Lengend Snippet: Fig. 2. Ab1–42 suppresses TRPV1 activation in mouse DRG neurons and in a capsaicin-induced acute pain model via LRP1: Electrophysiological and calcium imaging studies. a–c: Representative traces (a, b) and bar graph (c) showing 100 nM capsaicin-evoked inward currents after 1 lM Ab1–42 injection in mouse DRG (mDRG) neurons. **p < 0.01, unpaired t-test; control, n = 4; Ab1–42, n = 5. d–g: Representative traces (d–f) and bar graph (g) show a 100 nM capsaicin-induced Ca2+ increase after 1 lM Ab1–42 injection following 30 nM LRPAP1 pretreatment in mDRG neurons. ****, #### p < 0.0001, one-way ANOVA; control, n = 28; Ab1–42, n = 13; Ab1–42 + LRPAP1, n = 30. h, i: Hot plate and Hargreaves tests for basal heat sensitivity after intraplantar injection of 1 lg LRPAP1 in mature adult mice. *p < 0.05, two-way ANOVA with the Sidak post-hoc test, n = 7/group. g: Hargreaves test for basal heat sensitivity after intraplantar injection of 5 lg LRP1-neutralizing antibody in young mice. **p < 0.01, two-way ANOVA with Sidak post-hoc test; IgG, n = 5; LRP1 antibody, n = 7. Data are expressed as mean ± SEM. DRG, dorsal root ganglion; LRP1, low-density lipoprotein receptor-related protein 1; TRPV1, transient receptor potential vanilloid 1; LRPAP1, low-density lipoprotein receptor-related protein-associated protein 1; ANOVA, analysis of variance; SEM, standard error of the mean.

Article Snippet: Human TRPV1 was purchased from Origene (Rockville, MD, USA), and human SHP2-EGFP was purchased from Addgene (Watertown, MA, USA).

Techniques: Activation Assay, Imaging, Injection, Control

Journal: Journal of advanced research

Article Title: The LRP1-SHP2 pathway regulates TRPV1 sensitivity in the peripheral nervous system: Insights from amyloid beta 1-42 modulation.

doi: 10.1016/j.jare.2025.03.005

Figure Lengend Snippet: Fig. 3. LRP1 is co-expressed with TRPV1 in mouse DRG neurons: Immunohistochemical analysis. a: Immunohistochemistry showing LRP1 expression in DRG slices, n = 3 (NeuN: neuronal marker, NF200: large-sized-neuronal marker, CGRP: peptidergic small-sized neuronal marker, IB4: nonpeptidergic small-sized neuronal marker). Scale bar: 50 lm. b: Immunohistochemistry showing LRP1 and IB4 on L4-spinal cord sections, n = 3. Scale bar, 100 lm. c: In situ hybridization showing Lrp1 and Trpv1 expression in DRG slices, n = 3. Scale bar: 100 lm. d: Size frequency distribution of Lrp1 and Trpv1-positive and total neurons in mouse DRGs. e: Single-cell RT-PCR showing Lrp1 and Trpv1gene co-expression in mDRG neurons. Data are expressed as mean ± SEM. DRG, dorsal root ganglion; LRP1, low-density lipoprotein receptor-related protein 1; TRPV1, transient receptor potential vanilloid 1; CGRP, calcitonin gene-related peptide; IB4, isolectin B4; SEM, standard error of the mean.

Article Snippet: Human TRPV1 was purchased from Origene (Rockville, MD, USA), and human SHP2-EGFP was purchased from Addgene (Watertown, MA, USA).

Techniques: Immunohistochemical staining, Immunohistochemistry, Expressing, Marker, In Situ Hybridization, Reverse Transcription Polymerase Chain Reaction

Journal: Journal of advanced research

Article Title: The LRP1-SHP2 pathway regulates TRPV1 sensitivity in the peripheral nervous system: Insights from amyloid beta 1-42 modulation.

doi: 10.1016/j.jare.2025.03.005

Figure Lengend Snippet: Fig. 4. Knockdown of LRP1 abolishes inhibition of TRPV1 activation by Ab1–42 in mouse DRG neurons. a–c: Representative traces (a, b) and bar graph (c) showing 100 nM capsaicin-evoked inward currents after 1 lM Ab1–42 treatment in mouse DRG (mDRG) neurons. **p < 0.01, unpaired t-test; control, n = 4; Ab1–42, n = 5. d: Representative traces showing 1 lM capsaicin-evoked inward currents after 1 lM Ab1–42 treatment in TRPV1 (black)- and TRPV1 + LRP1 (turquoise)-expressing HEK293T cells. e: Bar graph showing the inward currents normalized by the first inward current. ***p < 0.001, unpaired t-test; TRPV1, n = 8; TRPV1/LRP1, n = 7. f–h: Representative traces (f, g) and a bar graph (h) show the 100 nM capsaicin-induced Ca2+ increase after 1 lM Ab1–42 treatment following 30 nM LRPAP1 pretreatment in mDRG neurons. ****, #### p < 0.0001, one- way ANOVA; control, n = 28; Ab1–42, n = 13; Ab1–42 + LRPAP1, n = 30. i: Representative traces showing the 1 lM capsaicin-induced Ca2+ increase after 1 lM Ab1–42 treatment in TRPV1 (black)- and TRPV1 + LRP1 (turquoise)-expressing HEK293T cells. j: Bar graph showing the Ca2+ increases normalized by the first Ca2+ increase. ****p < 0.0001, unpaired t-test; TRPV1, n = 12; TRPV1/LRP1, n = 34. Data are expressed as mean ± SEM. DRG, dorsal root ganglion; LRP1, low-density lipoprotein receptor-related protein 1; TRPV1, transient receptor potential vanilloid 1; LRPAP1, low-density lipoprotein receptor-related protein-associated protein 1; ANOVA, analysis of variance; SEM, standard error of the mean.

Article Snippet: Human TRPV1 was purchased from Origene (Rockville, MD, USA), and human SHP2-EGFP was purchased from Addgene (Watertown, MA, USA).

Techniques: Knockdown, Inhibition, Activation Assay, Control, Expressing

Journal: Journal of advanced research

Article Title: The LRP1-SHP2 pathway regulates TRPV1 sensitivity in the peripheral nervous system: Insights from amyloid beta 1-42 modulation.

doi: 10.1016/j.jare.2025.03.005

Figure Lengend Snippet: Fig. 5. Ab1–42 modulates TRPV1 activation via SHP2: calcium imaging, western blot, and electrophysiology studies. a–f: Representative traces (a–e) and a bar graph (f) showing the 100 nM capsaicin-induced Ca2+ increase after 1 lM Ab1–42 treatment following 1 lM SHP inhibitor (NSC-87877: pan-SHP inhibitor [green], PiPTⅢ: SHP1 inhibitor [blue], SHP099: SHP2 inhibitor [purple]) pretreatment in mouse DRG (mDRG) neurons. **p < 0.01, ## p < 0.01, ### p < 0.001, one-way ANOVA; control, n = 21; Ab1–42, n = 20; Ab1–42 + NSC87877, n = 30; Ab1–42 + PTPiIII, n = 12; Ab1–42 + SHP099, n = 17. g: Bands showing an increase of phospho-SHP2 (Y542) levels at 30 min after Ab1–42 (1 lM) treatment in LRP1-expressing HEK293T cells. h: Bar graph showing normalized phospho-SHP2 (Y542) intensity. **p < 0.01, unpaired t-test, n = 4/group. Data are expressed as mean ± SEM. i: Co-immunoprecipitation showing LRP1/SHP2 interaction in TRPV1/SHP2-expressing HEK293T cells. j–l: Representative traces (j–k) and a bar graph (l) showing 100 nM capsaicin-evoked inward currents after 1 lM SHP099 pretreatment in mDRG neurons. ****, #### p < 0.0001, one-way ANOVA; control, n = 6; Ab1–42, n = 5; Ab1–42 + SHP099, n = 7. DRG, dorsal root ganglion; LRP1, low-density lipoprotein receptor-related protein 1; TRPV1, transient receptor potential vanilloid 1; SHP2, src- homology domain-2–containing protein tyrosine phosphatase; ANOVA, analysis of variance; SEM, standard error of the mean. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Human TRPV1 was purchased from Origene (Rockville, MD, USA), and human SHP2-EGFP was purchased from Addgene (Watertown, MA, USA).

Techniques: Activation Assay, Imaging, Western Blot, Control, Expressing, Immunoprecipitation

Journal: Journal of advanced research

Article Title: The LRP1-SHP2 pathway regulates TRPV1 sensitivity in the peripheral nervous system: Insights from amyloid beta 1-42 modulation.

doi: 10.1016/j.jare.2025.03.005

Figure Lengend Snippet: Fig. 6. Ab1–42 modulates TRPV1 activation via SHP2 in human DRG neurons. a: Human DRG (hDRG) neurons loaded with Fura-2. Scale bar = 25 lm. b–d: Representative traces (b–c) and a bar graph (d) show a 1 lM capsaicin-induced Ca2+ increase after 1 lM Ab1–42 treatment in hDRG neurons. ***p < 0.001, unpaired t-test; control, n = 7; Ab1–

Article Snippet: Human TRPV1 was purchased from Origene (Rockville, MD, USA), and human SHP2-EGFP was purchased from Addgene (Watertown, MA, USA).

Techniques: Activation Assay, Control

Journal: Journal of advanced research

Article Title: The LRP1-SHP2 pathway regulates TRPV1 sensitivity in the peripheral nervous system: Insights from amyloid beta 1-42 modulation.

doi: 10.1016/j.jare.2025.03.005

Figure Lengend Snippet: Fig. 7. Ab1–42 modulates TRPV1 activation via SHP2. a–b: Representative traces (a) and a bar graph (b) showing the 100 nM capsaicin-evoked inward currents after 1 nM a2M injection in mouse DRG (mDRG) neurons. ***p < 0.001, unpaired t-test; control, n = 4; a2M, n = 5. c–d: Representative traces (c) and a bar graph (d) show the 100 nM capsaicin-induced Ca2+ increase after 1 nM a2M injection in mDRG neurons. ***p < 0.001, one-way ANOVA; control, n = 20; a2M, n = 20. e–f: Representative traces (e) and a bar graph (f) show the 1 lM capsaicin-induced Ca2+ increase after 1 nM a2M treatment in hDRG neurons. ****p < 0.0001, unpaired t-test; control, n = 7; a2M, n = 7. g–h: Representative traces (g) and a bar graph (h) showing the 100 nM capsaicin-induced Ca2+ increase after 1 nM a2M treatment following 1 lM SHP inhibitor (NSC-87877: pan- SHP inhibitor [green], SHP099: SHP2 inhibitor [purple]) pretreatment in mouse DRG (mDRG) neurons. # p < 0.05, *** p < 0.001, ### p < 0.001, one-way ANOVA; control, n = 20; a2M, n = 20; a2M + NSC87877, n = 20; a2M + SHP099, n = 20. DRG, dorsal root ganglion; TRPV1, transient receptor potential vanilloid 1; SHP2, src-homology domain-2– containing protein tyrosine phosphatase; a2M, a2-macroglobulin; ANOVA, analysis of variance. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Human TRPV1 was purchased from Origene (Rockville, MD, USA), and human SHP2-EGFP was purchased from Addgene (Watertown, MA, USA).

Techniques: Activation Assay, Injection, Control

Journal: Journal of advanced research

Article Title: The LRP1-SHP2 pathway regulates TRPV1 sensitivity in the peripheral nervous system: Insights from amyloid beta 1-42 modulation.

doi: 10.1016/j.jare.2025.03.005

Figure Lengend Snippet: Fig. 8. Ab1–42 and a2M modulate heat hyperalgesia in an SNI model via the LRP1-SHP2 pathway. a: Nociceptive responses evoked by intraplantar injection of 0.125 lg capsaicin with 1 lg Ab1–42 and 0.5 lg LRPAP1. # p < 0.05, **, ## p < 0.01, ****, #### p < 0.0001, two-way ANOVA with the Sidak post-hoc test. b: Schematic illustration of the spared nerve injury (SNI) model used to induce neuropathic pain in mice. c: Bands showing LRP1 protein expression after injection of AAV5-control (black) and AAV5-cre (orange) in mouse DRG. d: The bar graph shows normalized band intensity (n = 4). **p < 0.01. e: Heat hyperalgesia in the SNI chronic pain model following intraplantar injection of 1 lg Ab1–42 and 0.5 lg LRPAP1. # p < 0.05, **, ## p < 0.01, ***p < 0.001, two-way ANOVA with the Sidak post-hoc test; vehicle, n = 4; Ab1–42, n = 7; Ab1–42 + LRPAP1, n = 8. f: SNI-induced heat hyperalgesia in control and LRP1 cKO mice following intraplantar injection of 1 lg Ab1–42. **p < 0.01, two-way ANOVA with the Sidak post-hoc test; control, n = 6; LRP1 cKO, n = 7. g: Heat hyperalgesia in the SNI chronic pain model following intraplantar injection of 1 lg Ab1–42 and 2 lg SHP099. ## p < 0.01, ### p < 0.001, ****, #### p < 0.0001, two-way ANOVA with the Sidak post-hoc test; vehicle, n = 6; Ab1–42, n = 10; Ab1–42 + SHP099, n = 9. Data are expressed as mean ± SEM. h: Heat hyperalgesia in the SNI neuropathic pain model following intraplantar a2M and LRPAP1 (0.5 lg) injection. i: Heat hyperalgesia in TRPV1-cre;LRP1flox/flox mice following intraplantar a2M injection after SNI surgery. j: Heat hyperalgesia assessed by the Hargreaves test in the SNI chronic pain model following intraplantar a2M and SHP099 (2 lg) injection. k: Schematic of LRP1-induced TRPV1 inhibition via SHP2 activation: The role of Ab1–42 and a2M. Data are shown as the mean ± SEM. DRG, dorsal root ganglion; LRP1, low-density lipoprotein receptor-related protein 1; TRPV1, transient receptor potential vanilloid 1; LRPAP1, low-density lipoprotein receptor-related protein-associated protein 1; SHP2, src-homology domain-2–containing protein tyrosine phosphatase; a2M, a2-macroglobulin; cKO, conditional knockout; ANOVA, analysis of variance; SEM, standard error of the mean. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Human TRPV1 was purchased from Origene (Rockville, MD, USA), and human SHP2-EGFP was purchased from Addgene (Watertown, MA, USA).

Techniques: Injection, Expressing, Control, Inhibition, Activation Assay, Knock-Out