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Millipore pacap38 peptide

(A) Western blotting analysis of AR55Q in HEK293T cells treated with PACAP and peptides 2, 4, and 7. Quantification is shown at the bottom. n = 3 to 5 independent experiments; *P < 0.05. (B) XTT [2,3-bis (2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino) carbonyl]-2H-tetrazolium hydroxide] assay in MN-1 cells expressing AR65Q treated with peptides 2, 4, and 7 and 100 nM PACAP for 48 hours. n = 3 independent experiments. Veh, vehicle. (C) Real-time polymerase chain reaction (PCR) analysis of the transcript levels of Pac1, Vpac1, and Vpac2 receptors in the spinal cord and skeletal muscle of 180-day-old control (CTR; wild type) and AR113Q mice and normalized to hypoxanthine phosphoribosyltransferase 1 (Hprt1) and Pgk1 in spinal cord and to Hprt1 and β-glucuronidase (Gusβ) in skeletal muscle. n = 6 mice for each group. (D) cAMP level analysis in the spinal cord of control mice treated with PACAP27, <t>PACAP38,</t> peptide 2, and peptide 7. n = 3 mice for each group. Graphs, means ± SEM; one-way ANOVA was used.

Pacap38 Peptide, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/pacap38 peptide/product/Millipore
Average 90 stars, based on 1 article reviews

pacap38 peptide - by Bioz Stars, 2026-03

90/100 stars

Images

1) Product Images from "Adenylyl cyclase activating polypeptide reduces phosphorylation and toxicity of the polyglutamine-expanded androgen receptor in spinobulbar muscular atrophy"

Article Title: Adenylyl cyclase activating polypeptide reduces phosphorylation and toxicity of the polyglutamine-expanded androgen receptor in spinobulbar muscular atrophy

Journal: Science translational medicine

doi: 10.1126/scitranslmed.aaf9526

Figure Legend Snippet: (A) Western blotting analysis of AR55Q in HEK293T cells treated with PACAP and peptides 2, 4, and 7. Quantification is shown at the bottom. n = 3 to 5 independent experiments; *P < 0.05. (B) XTT [2,3-bis (2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino) carbonyl]-2H-tetrazolium hydroxide] assay in MN-1 cells expressing AR65Q treated with peptides 2, 4, and 7 and 100 nM PACAP for 48 hours. n = 3 independent experiments. Veh, vehicle. (C) Real-time polymerase chain reaction (PCR) analysis of the transcript levels of Pac1, Vpac1, and Vpac2 receptors in the spinal cord and skeletal muscle of 180-day-old control (CTR; wild type) and AR113Q mice and normalized to hypoxanthine phosphoribosyltransferase 1 (Hprt1) and Pgk1 in spinal cord and to Hprt1 and β-glucuronidase (Gusβ) in skeletal muscle. n = 6 mice for each group. (D) cAMP level analysis in the spinal cord of control mice treated with PACAP27, PACAP38, peptide 2, and peptide 7. n = 3 mice for each group. Graphs, means ± SEM; one-way ANOVA was used.

Techniques Used: Western Blot, Expressing, Real-time Polymerase Chain Reaction, Control

Image Search Results

A A schematic coronal section from the rat brain atlas illustrating cannula implantation into the right lateral ventricle (left panel) and the experimental timeline (right panel) ( B ) PACAP38-injected animals ( n = 8) showed a significant reduction in active coping behavior, as indicated by decreased struggling, and an enhancement of immobility (floating behavior) compared to aCSF-injected controls ( n = 6). Data are expressed as mean ± SEM. * p < 0.05, ** p < 0.01 vs aCSF-injected controls (Student's t- test unpaired).

Journal: Neuropsychopharmacology

Article Title: PACAP regulates neuroendocrine and behavioral stress responses via CRF-containing neurons of the rat hypothalamic paraventricular nucleus

doi: 10.1038/s41386-024-02016-9

Figure Lengend Snippet: A A schematic coronal section from the rat brain atlas illustrating cannula implantation into the right lateral ventricle (left panel) and the experimental timeline (right panel) ( B ) PACAP38-injected animals ( n = 8) showed a significant reduction in active coping behavior, as indicated by decreased struggling, and an enhancement of immobility (floating behavior) compared to aCSF-injected controls ( n = 6). Data are expressed as mean ± SEM. * p < 0.05, ** p < 0.01 vs aCSF-injected controls (Student's t- test unpaired).

Article Snippet: PACAP38 (Bachem AG, Switzerland) were dissolved in sterile distilled water and aliquots of stock solution (1 mg/mL) were stored at −80 °C.

Techniques: Injection

A Schematic coronal sections from the brain atlas showing brain regions examined (gray shaded). B Representative photomicrographs showing c-Fos positive cells in rats exposed to forced swim stress and microinjected either with aCSF or PACAP38. The lower panels show c-Fos quantification presented as bar graphs in the PVN ( C ) LS ( D ) and BNST ( E ). There was a significant increase of stress-induced c-Fos expression in the parvocellular part of the PVN, ventral LS and latero-dorsal part of the BNST of PACAP38-treated animals compared to aCSF-injected controls. Abbreviations: 3 V third ventricle, AC anterior commissure, BNST bed nucleus of the stria terminalis, BNSTld latero-dorsal part of the BNST, BNSTlp latero-posterior part of the BNST, BNSTma medial-anterior part of the BNST, CC corpus callosum, LS lateral septum, LSd lateral septum dorsal, LSv lateral septum ventral, LV lateral ventricle, opt optic nerve, PVN paraventricular nucleus of the hypothalamus, pcPVN parvocellular part of the PVN, mcPVN magnocellular part of the PVN. N = 6–8 animals per group. Scale bars: 100 µm at lower magnification, 25 µm at higher magnification images. Data are expressed as mean ± SEM. * p < 0.05 vs aCSF-injected controls (Student’s t- test).

Journal: Neuropsychopharmacology

Article Title: PACAP regulates neuroendocrine and behavioral stress responses via CRF-containing neurons of the rat hypothalamic paraventricular nucleus

doi: 10.1038/s41386-024-02016-9

Figure Lengend Snippet: A Schematic coronal sections from the brain atlas showing brain regions examined (gray shaded). B Representative photomicrographs showing c-Fos positive cells in rats exposed to forced swim stress and microinjected either with aCSF or PACAP38. The lower panels show c-Fos quantification presented as bar graphs in the PVN ( C ) LS ( D ) and BNST ( E ). There was a significant increase of stress-induced c-Fos expression in the parvocellular part of the PVN, ventral LS and latero-dorsal part of the BNST of PACAP38-treated animals compared to aCSF-injected controls. Abbreviations: 3 V third ventricle, AC anterior commissure, BNST bed nucleus of the stria terminalis, BNSTld latero-dorsal part of the BNST, BNSTlp latero-posterior part of the BNST, BNSTma medial-anterior part of the BNST, CC corpus callosum, LS lateral septum, LSd lateral septum dorsal, LSv lateral septum ventral, LV lateral ventricle, opt optic nerve, PVN paraventricular nucleus of the hypothalamus, pcPVN parvocellular part of the PVN, mcPVN magnocellular part of the PVN. N = 6–8 animals per group. Scale bars: 100 µm at lower magnification, 25 µm at higher magnification images. Data are expressed as mean ± SEM. * p < 0.05 vs aCSF-injected controls (Student’s t- test).

Article Snippet: PACAP38 (Bachem AG, Switzerland) were dissolved in sterile distilled water and aliquots of stock solution (1 mg/mL) were stored at −80 °C.

Techniques: Expressing, Injection

A Schematic drawings of coronal sections of the rat brain showing the localization of the cannula tips within the PVN from rostral (−1.6 mm) to caudal (−1.88 mm) for bilateral microinjection of vehicle (white circles) and PACAP38 (black circles). B Intra-PVN microinjections of PACAP38 (150 and 15 pmol/site; n = 10 and 4) elicited significantly higher floating (passive coping) and reduced struggling (active coping) time compared to aCSF-injected controls ( n = 6). No significant effects were found on the swimming behavior between PACAP38-injected rats and controls. Data are expressed as mean ± SEM. * p < 0.05 ** p < 0.01 vs aCSF-injected controls (one-way ANOVA followed by Dunnet's multiple comparison post hoc test).

Journal: Neuropsychopharmacology

Article Title: PACAP regulates neuroendocrine and behavioral stress responses via CRF-containing neurons of the rat hypothalamic paraventricular nucleus

doi: 10.1038/s41386-024-02016-9

Figure Lengend Snippet: A Schematic drawings of coronal sections of the rat brain showing the localization of the cannula tips within the PVN from rostral (−1.6 mm) to caudal (−1.88 mm) for bilateral microinjection of vehicle (white circles) and PACAP38 (black circles). B Intra-PVN microinjections of PACAP38 (150 and 15 pmol/site; n = 10 and 4) elicited significantly higher floating (passive coping) and reduced struggling (active coping) time compared to aCSF-injected controls ( n = 6). No significant effects were found on the swimming behavior between PACAP38-injected rats and controls. Data are expressed as mean ± SEM. * p < 0.05 ** p < 0.01 vs aCSF-injected controls (one-way ANOVA followed by Dunnet's multiple comparison post hoc test).

Article Snippet: PACAP38 (Bachem AG, Switzerland) were dissolved in sterile distilled water and aliquots of stock solution (1 mg/mL) were stored at −80 °C.

Techniques: Microinjection, Injection, Comparison

A Schematic illustration of the experimental design with the timeline of blood sampling (red circles), drug infusion (green bar) and stress exposure (orange bar, forced swim, FS). Experiment started with insertion of the infusion device (bilateral injection cannulas connected to a microinfusion pump) at least 1 h before blood sampling started. Drugs were infused automatically at a constant flow rate over a period of 7.5 min without any stressful manipulations (e.g such as capturing or restraining animals) before and during the infusion procedure. Blood samples were collected at regular intervals before drug infusion under basal conditions (−35 and −15 min) and after drug infusion, but before stress exposure (−1 min) and after forced swim stress (10, 30, and 60 min). B Swim stress caused an increase in plasma ACTH levels in both intra-PVN PACAP38 ( n = 7) and aCSF-injected controls ( n = 7). Compared to controls, intra-PVN PACAP38-injected rats showed higher plasma ACTH levels during and after forced swim stress. However, basal levels did not differ between groups. The green bar indicates timing of intra-PVN infusion, the orange bar the forced swim (FS) stress exposure. Data are expressed as mean ± SEM. * p < 0.05 **** p < 0.0001 compared to basal timepoints (−35 and −15 min) in same treatment group; +++ p < 0.001 compared to vehicle-injected controls at same timepoint (two-way ANOVA followed by Bonferroni's multiple comparison post hoc test).

Journal: Neuropsychopharmacology

Article Title: PACAP regulates neuroendocrine and behavioral stress responses via CRF-containing neurons of the rat hypothalamic paraventricular nucleus

doi: 10.1038/s41386-024-02016-9

Figure Lengend Snippet: A Schematic illustration of the experimental design with the timeline of blood sampling (red circles), drug infusion (green bar) and stress exposure (orange bar, forced swim, FS). Experiment started with insertion of the infusion device (bilateral injection cannulas connected to a microinfusion pump) at least 1 h before blood sampling started. Drugs were infused automatically at a constant flow rate over a period of 7.5 min without any stressful manipulations (e.g such as capturing or restraining animals) before and during the infusion procedure. Blood samples were collected at regular intervals before drug infusion under basal conditions (−35 and −15 min) and after drug infusion, but before stress exposure (−1 min) and after forced swim stress (10, 30, and 60 min). B Swim stress caused an increase in plasma ACTH levels in both intra-PVN PACAP38 ( n = 7) and aCSF-injected controls ( n = 7). Compared to controls, intra-PVN PACAP38-injected rats showed higher plasma ACTH levels during and after forced swim stress. However, basal levels did not differ between groups. The green bar indicates timing of intra-PVN infusion, the orange bar the forced swim (FS) stress exposure. Data are expressed as mean ± SEM. * p < 0.05 **** p < 0.0001 compared to basal timepoints (−35 and −15 min) in same treatment group; +++ p < 0.001 compared to vehicle-injected controls at same timepoint (two-way ANOVA followed by Bonferroni's multiple comparison post hoc test).

Article Snippet: PACAP38 (Bachem AG, Switzerland) were dissolved in sterile distilled water and aliquots of stock solution (1 mg/mL) were stored at −80 °C.

Techniques: Sampling, Injection, Clinical Proteomics, Comparison

A Schematic drawing illustrating different subregions of the PVN and representative confocal images showing CRF (red) and c-Fos (blue) immunopositive neurons in the mpcPVN. B The lower panels show quantification of CRF (left panel) and CRF/c-Fos (middle and right panel) positive neurons presented as bar graphs in the mpcPVN. There was no significant difference in the total number of CRF neurons between PACAP38-treated animals and controls, but the number of CRF neurons that co-express c-Fos were significantly higher in the PACAP38-treated animals compared to controls. Note that in controls only 38% of CRF neurons expressed Fos, while in the PACAP38-treated animals the percentage of CRF neurons expressing c-Fos was 76%. Abbreviations: 3 V third ventricle, ns not significant. N = 3 animals per group. Scale bar: 50 µm. Data are expressed as mean ± SEM. * p < 0.05 ** p < 0.01 vs aCSF-injected controls (Student's t- test).

Journal: Neuropsychopharmacology

Article Title: PACAP regulates neuroendocrine and behavioral stress responses via CRF-containing neurons of the rat hypothalamic paraventricular nucleus

doi: 10.1038/s41386-024-02016-9

Figure Lengend Snippet: A Schematic drawing illustrating different subregions of the PVN and representative confocal images showing CRF (red) and c-Fos (blue) immunopositive neurons in the mpcPVN. B The lower panels show quantification of CRF (left panel) and CRF/c-Fos (middle and right panel) positive neurons presented as bar graphs in the mpcPVN. There was no significant difference in the total number of CRF neurons between PACAP38-treated animals and controls, but the number of CRF neurons that co-express c-Fos were significantly higher in the PACAP38-treated animals compared to controls. Note that in controls only 38% of CRF neurons expressed Fos, while in the PACAP38-treated animals the percentage of CRF neurons expressing c-Fos was 76%. Abbreviations: 3 V third ventricle, ns not significant. N = 3 animals per group. Scale bar: 50 µm. Data are expressed as mean ± SEM. * p < 0.05 ** p < 0.01 vs aCSF-injected controls (Student's t- test).

Article Snippet: PACAP38 (Bachem AG, Switzerland) were dissolved in sterile distilled water and aliquots of stock solution (1 mg/mL) were stored at −80 °C.

Techniques: Expressing, Injection

Journal: Science translational medicine

Article Title: Adenylyl cyclase activating polypeptide reduces phosphorylation and toxicity of the polyglutamine-expanded androgen receptor in spinobulbar muscular atrophy

doi: 10.1126/scitranslmed.aaf9526

Figure Lengend Snippet: (A) Western blotting analysis of AR55Q in HEK293T cells treated with PACAP and peptides 2, 4, and 7. Quantification is shown at the bottom. n = 3 to 5 independent experiments; *P < 0.05. (B) XTT [2,3-bis (2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino) carbonyl]-2H-tetrazolium hydroxide] assay in MN-1 cells expressing AR65Q treated with peptides 2, 4, and 7 and 100 nM PACAP for 48 hours. n = 3 independent experiments. Veh, vehicle. (C) Real-time polymerase chain reaction (PCR) analysis of the transcript levels of Pac1, Vpac1, and Vpac2 receptors in the spinal cord and skeletal muscle of 180-day-old control (CTR; wild type) and AR113Q mice and normalized to hypoxanthine phosphoribosyltransferase 1 (Hprt1) and Pgk1 in spinal cord and to Hprt1 and β-glucuronidase (Gusβ) in skeletal muscle. n = 6 mice for each group. (D) cAMP level analysis in the spinal cord of control mice treated with PACAP27, PACAP38, peptide 2, and peptide 7. n = 3 mice for each group. Graphs, means ± SEM; one-way ANOVA was used.

Article Snippet: Mice were treated with vehicle [intranasal solution containing 7.5 mg of NaCl, 1.7 mg of citric acid monohydrate, 3 mg of disodium phosphate dihydrate, and 0.2 mg of benzalkonium chloride solution (50%) per milliliter; PACAP38 (137061-48-4, Calbiochem); or peptide 7 (2, 10, and 20 μg) dissolved in vehicle ( 51 )].

Techniques: Western Blot, Expressing, Real-time Polymerase Chain Reaction, Control

PACAP38-MrgprB2 pathway mediates stress-induced headache behaviours. a Facial mechanical withdrawal thresholds at consecutive time points after restraint stress (1 h/day for 3 days) and PACAP38 dural injection (0.1 ng) in wild-type (WT) or MrgprB2-deficient (KO) mice ( n = 5 WT CON, 6 WT STR, and 9 KO STR mice). b Blood concentration of PACAP38 ( n = 6 mice per group). Blood was collected immediately after cessation of stress or 24 h after stress. c β-hexosaminidase release by mouse peritoneal mast cells ( n = 6 wells per group). d Facial mechanical withdrawal thresholds after dural injection with PACAP38 (1 ng) or IL-6 (0.1 ng) in WT or KO mice ( n = 7 WT Veh, 7 WT PACAP, 5 WT IL-6, 6 KO Veh, and 6 KO PACAP mice). e Grimace score ( n = 6 mice per group). ( right ) Representative mouse facial expression images. CON: control, C48/80: compound 48/80, STR: stress, IL-6: interleukin-6, Veh: vehicle. Significance comparisons: (a) *WT STR vs. WT CON; + WT STR vs. KO STR; (d, e) *WT Veh. vs. WT PACAP38; + WT PACAP38 vs. KO PACAP38. Error bars indicate S.E.M. * /+ p < 0.05; ** /++ p < 0.01; *** /+++ p < 0.001; (a, d, and e) two-way ANOVA with Tukey’s multiple comparison post-hoc test; (b and c) one-way ANOVA with Tukey’s multiple comparison post-hoc test

Journal: The Journal of Headache and Pain

Article Title: PACAP38/mast-cell-specific receptor axis mediates repetitive stress-induced headache in mice

doi: 10.1186/s10194-024-01786-3

Figure Lengend Snippet: PACAP38-MrgprB2 pathway mediates stress-induced headache behaviours. a Facial mechanical withdrawal thresholds at consecutive time points after restraint stress (1 h/day for 3 days) and PACAP38 dural injection (0.1 ng) in wild-type (WT) or MrgprB2-deficient (KO) mice ( n = 5 WT CON, 6 WT STR, and 9 KO STR mice). b Blood concentration of PACAP38 ( n = 6 mice per group). Blood was collected immediately after cessation of stress or 24 h after stress. c β-hexosaminidase release by mouse peritoneal mast cells ( n = 6 wells per group). d Facial mechanical withdrawal thresholds after dural injection with PACAP38 (1 ng) or IL-6 (0.1 ng) in WT or KO mice ( n = 7 WT Veh, 7 WT PACAP, 5 WT IL-6, 6 KO Veh, and 6 KO PACAP mice). e Grimace score ( n = 6 mice per group). ( right ) Representative mouse facial expression images. CON: control, C48/80: compound 48/80, STR: stress, IL-6: interleukin-6, Veh: vehicle. Significance comparisons: (a) *WT STR vs. WT CON; + WT STR vs. KO STR; (d, e) *WT Veh. vs. WT PACAP38; + WT PACAP38 vs. KO PACAP38. Error bars indicate S.E.M. * /+ p < 0.05; ** /++ p < 0.01; *** /+++ p < 0.001; (a, d, and e) two-way ANOVA with Tukey’s multiple comparison post-hoc test; (b and c) one-way ANOVA with Tukey’s multiple comparison post-hoc test

Article Snippet: Animals were anesthetized with Ketamine/Xylazine (80/10 mg/kg) 3 h after PACAP38 injection (MilliporeSigma, St Louis, MO, USA), and cardiac perfusion fixation was conducted using 4% paraformaldehyde.

Techniques: Injection, Concentration Assay, Expressing, Comparison

PACAP38 activates MrgprB2/MRGPRX2 and induces mast cell degranulation in dura mater for trigeminovascular alternations. a and b ( left ) Representative Ca 2+ fluorescence images showing increase in HEK293 cells expressing MrgprB2 (a) or MRGPRX2 (b) with PACAP38 application (1 µM). ( right , top ) Percentage of cells activated by PACAP38 in MrgprB2-transfected cells ( n = 13 cultures) (a) or MRGPRX2-transfected cells ( n = 20 cultures) (b). ( right , bottom ) Average Ca 2+ transient trace of cells activated by PACAP38 from MrgprB2 (10 µM)-transfected cells (a) or MRGPRX2 (1 µM)-transfected cells (b). c and e ( left ) Representative Ca 2+ fluorescent images showing increase by PACAP38 (50 µM) in mouse mast cells (c) or LAD2 cells (e). ( right , top ) Percentage of total cells activated by PACAP38 from mouse mast cells ( n = 13 cultures) (c) or LAD2 cells ( n = 12 cultures) (e). ( right , bottom ) Average Ca 2+ transient trace of mouse mast cells (c) or LAD2 cells activated by PACAP38 (e). d Percentage of mast cells derived from wild type (WT) and MrgprB2-deficient (KO) mice activated by PACAP38 (10 µM) ( n = 7 cultures). f Percentage of LAD2 cells activated by PACAP38 (1 µM) and treated with vehicle or QWF ( n = 6 cultures). g ( left ) Representative confocal fluorescence images of avidin-stained (red) dura mater mast cells. ( right ) Percentage of degranulated mast cells ( n = 4 mice per group). h ( left ) in vivo dura Pirt -GCaMP3 Ca 2+ imaging in trigeminal nerve afferents with PACAP38 application. ( right ) Relative change of Ca 2+ transient with PACAP38 (10 µM) through cranial window ( n = 4 WT and 6 KO mice). i ( left ) Representative images showing in vivo fluorescence of dura blood vessels before and after PACAP38 (10 µM) application through cranial window. ( right ) Percentage change in blood vessel diameter after PACAP38 application ( n = 4 WT and 6 KO mice). Error bars indicate S.E.M. * p < 0.05; ** p < 0.01; *** p < 0.001; (d, f, h, i) two-tailed Student’s t -test, (g) one-way ANOVA with Tukey’s multiple comparison post-hoc test

Journal: The Journal of Headache and Pain

Article Title: PACAP38/mast-cell-specific receptor axis mediates repetitive stress-induced headache in mice

doi: 10.1186/s10194-024-01786-3

Figure Lengend Snippet: PACAP38 activates MrgprB2/MRGPRX2 and induces mast cell degranulation in dura mater for trigeminovascular alternations. a and b ( left ) Representative Ca 2+ fluorescence images showing increase in HEK293 cells expressing MrgprB2 (a) or MRGPRX2 (b) with PACAP38 application (1 µM). ( right , top ) Percentage of cells activated by PACAP38 in MrgprB2-transfected cells ( n = 13 cultures) (a) or MRGPRX2-transfected cells ( n = 20 cultures) (b). ( right , bottom ) Average Ca 2+ transient trace of cells activated by PACAP38 from MrgprB2 (10 µM)-transfected cells (a) or MRGPRX2 (1 µM)-transfected cells (b). c and e ( left ) Representative Ca 2+ fluorescent images showing increase by PACAP38 (50 µM) in mouse mast cells (c) or LAD2 cells (e). ( right , top ) Percentage of total cells activated by PACAP38 from mouse mast cells ( n = 13 cultures) (c) or LAD2 cells ( n = 12 cultures) (e). ( right , bottom ) Average Ca 2+ transient trace of mouse mast cells (c) or LAD2 cells activated by PACAP38 (e). d Percentage of mast cells derived from wild type (WT) and MrgprB2-deficient (KO) mice activated by PACAP38 (10 µM) ( n = 7 cultures). f Percentage of LAD2 cells activated by PACAP38 (1 µM) and treated with vehicle or QWF ( n = 6 cultures). g ( left ) Representative confocal fluorescence images of avidin-stained (red) dura mater mast cells. ( right ) Percentage of degranulated mast cells ( n = 4 mice per group). h ( left ) in vivo dura Pirt -GCaMP3 Ca 2+ imaging in trigeminal nerve afferents with PACAP38 application. ( right ) Relative change of Ca 2+ transient with PACAP38 (10 µM) through cranial window ( n = 4 WT and 6 KO mice). i ( left ) Representative images showing in vivo fluorescence of dura blood vessels before and after PACAP38 (10 µM) application through cranial window. ( right ) Percentage change in blood vessel diameter after PACAP38 application ( n = 4 WT and 6 KO mice). Error bars indicate S.E.M. * p < 0.05; ** p < 0.01; *** p < 0.001; (d, f, h, i) two-tailed Student’s t -test, (g) one-way ANOVA with Tukey’s multiple comparison post-hoc test

Techniques: Fluorescence, Expressing, Transfection, Derivative Assay, Avidin-Biotin Assay, Staining, In Vivo, Imaging, Two Tailed Test, Comparison

Activation of the PACAP38-MrgprB2 pathway increases the number of spontaneously activated TG neurons assessed by in vivo TG Pirt -GCaMP Ca 2+ imaging. a Representative images of spontaneous in vivo activity assessed by TG Pirt -GCaMP3 Ca 2+ imaging after PACAP38 dural injection. V1 (ophthalmic), V2 (maxillary), and V3 (mandibular) indicate the location of neuronal cell bodies in TG imaging. White arrowheads indicate Spontaneous Ca 2+ activated neurons. b and d Representative heatmaps of spontaneously activated individual neurons in stressed (b) and PACAP38-injected (d) mice. c and e Number of total, small, medium, and large spontaneously activated neurons of each group in stressed (c) or PACAP38 injected (e) mice. (c) n = 6 WT CON, 6 WT STR, and 5 KO STR. (e) n = 6 mice per group. Small-diameter TG neurons (< 20 μm); medium (20–25 μm); large (> 25 μm). CON: control, KO: knock-out, STR: stress, Veh: vehicle, WT: wild-type. Error bars indicate S.E.M. ** p < 0.01; *** p < 0.001; one-way ANOVA with Tukey’s multiple comparison post-hoc test

Journal: The Journal of Headache and Pain

Article Title: PACAP38/mast-cell-specific receptor axis mediates repetitive stress-induced headache in mice

doi: 10.1186/s10194-024-01786-3

Figure Lengend Snippet: Activation of the PACAP38-MrgprB2 pathway increases the number of spontaneously activated TG neurons assessed by in vivo TG Pirt -GCaMP Ca 2+ imaging. a Representative images of spontaneous in vivo activity assessed by TG Pirt -GCaMP3 Ca 2+ imaging after PACAP38 dural injection. V1 (ophthalmic), V2 (maxillary), and V3 (mandibular) indicate the location of neuronal cell bodies in TG imaging. White arrowheads indicate Spontaneous Ca 2+ activated neurons. b and d Representative heatmaps of spontaneously activated individual neurons in stressed (b) and PACAP38-injected (d) mice. c and e Number of total, small, medium, and large spontaneously activated neurons of each group in stressed (c) or PACAP38 injected (e) mice. (c) n = 6 WT CON, 6 WT STR, and 5 KO STR. (e) n = 6 mice per group. Small-diameter TG neurons (< 20 μm); medium (20–25 μm); large (> 25 μm). CON: control, KO: knock-out, STR: stress, Veh: vehicle, WT: wild-type. Error bars indicate S.E.M. ** p < 0.01; *** p < 0.001; one-way ANOVA with Tukey’s multiple comparison post-hoc test

Techniques: Activation Assay, In Vivo, Imaging, Activity Assay, Injection, Knock-Out, Comparison

Activation of PACAP38-MrgprB2 pathway sensitizes TG neurons. a Representative images of in vivo TG neurons activated by 0.4 g von Frey filament application onto V1 region in PACAP38-injected mice. V1 (ophthalmic), V2 (maxillary), and V3 (mandibular) indicate the location of neuron cell bodies in TG imaging. White arrowheads indicate activated neurons. b and c Number of individual TG neurons activated by (b) 0.4 g von Frey filament or (c) capsaicin in stressed mice ( n = 6 WT CON, 6 WT STR, and 5 KO STR). d and e Number of individual TG neurons activated by (d) 0.4 g von Frey filament or (e) capsaicin in PACAP38-injected mice ( n = 6 mice per group). Small-diameter TG neurons (< 20 μm); medium (20–25 μm); large (> 25 μm). CON: control, KO: knock-out, STR: stress, Veh: vehicle, WT: wild-type. Error bars indicate S.E.M. * p < 0.05; ** p < 0.01; *** p < 0.001; (b-e) one-way ANOVA with Tukey’s multiple comparison post-hoc test

Journal: The Journal of Headache and Pain

Article Title: PACAP38/mast-cell-specific receptor axis mediates repetitive stress-induced headache in mice

doi: 10.1186/s10194-024-01786-3

Figure Lengend Snippet: Activation of PACAP38-MrgprB2 pathway sensitizes TG neurons. a Representative images of in vivo TG neurons activated by 0.4 g von Frey filament application onto V1 region in PACAP38-injected mice. V1 (ophthalmic), V2 (maxillary), and V3 (mandibular) indicate the location of neuron cell bodies in TG imaging. White arrowheads indicate activated neurons. b and c Number of individual TG neurons activated by (b) 0.4 g von Frey filament or (c) capsaicin in stressed mice ( n = 6 WT CON, 6 WT STR, and 5 KO STR). d and e Number of individual TG neurons activated by (d) 0.4 g von Frey filament or (e) capsaicin in PACAP38-injected mice ( n = 6 mice per group). Small-diameter TG neurons (< 20 μm); medium (20–25 μm); large (> 25 μm). CON: control, KO: knock-out, STR: stress, Veh: vehicle, WT: wild-type. Error bars indicate S.E.M. * p < 0.05; ** p < 0.01; *** p < 0.001; (b-e) one-way ANOVA with Tukey’s multiple comparison post-hoc test

Techniques: Activation Assay, In Vivo, Injection, Imaging, Knock-Out, Comparison

PACAP38 infusion and stress cause mechanical hypersensitivity in periorbital region via TNF-α receptor and TRPV1. a-d Head withdrawal thresholds tested by von Frey filament following restraint stress or PACAP38 dural injection. Mice were treated with i.p. injection of TRPV1-inhibitor (SB366791; 1 mg/kg) or TNF-α inhibitor (R-7050; 10 mg/kg) i.p. injection; or vehicle. (a) Withdrawal threshold in stressed mice treated with SB366791 ( n = 6 mice per group). (b) Threshold in stressed mice treated with R-7050 ( n = 5 mice per group). (c) Threshold in PACAP38-injected mice treated with SB366791 ( n = 6 mice per group) *(black) Veh/Veh vs. PACAP38/Veh; *(red) PACAP38/Veh vs. PACAP38/SB366791. (d) Threshold in PACAP38-injected mice treated with R-7050 ( n = 6 mice per group). Veh: vehicle. Error bars indicate S.E.M. * p < 0.05; ** p < 0.01; *** p < 0.001; (a, b, d) two-tailed Student’s t -test, (c) two-way ANOVA with Tukey’s multiple comparison post-hoc test

Journal: The Journal of Headache and Pain

Article Title: PACAP38/mast-cell-specific receptor axis mediates repetitive stress-induced headache in mice

doi: 10.1186/s10194-024-01786-3

Figure Lengend Snippet: PACAP38 infusion and stress cause mechanical hypersensitivity in periorbital region via TNF-α receptor and TRPV1. a-d Head withdrawal thresholds tested by von Frey filament following restraint stress or PACAP38 dural injection. Mice were treated with i.p. injection of TRPV1-inhibitor (SB366791; 1 mg/kg) or TNF-α inhibitor (R-7050; 10 mg/kg) i.p. injection; or vehicle. (a) Withdrawal threshold in stressed mice treated with SB366791 ( n = 6 mice per group). (b) Threshold in stressed mice treated with R-7050 ( n = 5 mice per group). (c) Threshold in PACAP38-injected mice treated with SB366791 ( n = 6 mice per group) *(black) Veh/Veh vs. PACAP38/Veh; *(red) PACAP38/Veh vs. PACAP38/SB366791. (d) Threshold in PACAP38-injected mice treated with R-7050 ( n = 6 mice per group). Veh: vehicle. Error bars indicate S.E.M. * p < 0.05; ** p < 0.01; *** p < 0.001; (a, b, d) two-tailed Student’s t -test, (c) two-way ANOVA with Tukey’s multiple comparison post-hoc test

Techniques: Injection, Two Tailed Test, Comparison

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