Journal: PLoS ONE

Article Title: Hydrogen Sulfide Plays a Key Role in the Inhibitory Neurotransmission to the Pig Intravesical Ureter

doi: 10.1371/journal.pone.0113580

Figure Lengend Snippet: Arrival of action potentials (AP) at the motor nerve terminal (MNT) nerve ending evokes membrane depolarization and activation of voltage-gated Ca 2+ (VOC) channels with the subsequent Ca 2+ influx, which would stimulate neuronal cystathionine γ-lyase (CSE), through interaction with calmodulin (CaM) favouring H 2 S synthesis from L-cysteine and release from nerves. H 2 S would diffuse across the synaptic cleft producing postjunctional K ATP channel activation, membrane hyperpolarization by K + efflux and subsequent smooth muscle (SM) relaxation. In addition, H 2 S might promote TRPA 1 and TRPV 1 channel activation from capsaicin-sensitive primary afferent (CSPA), thus favouring the release of pituitary adenylyl cyclase-activating polypeptide 38 (PACAP 38) and calcitonin gene-related peptide (CGRP), which in turns would produce smooth muscle relaxation via activation the VIP/PACAP (VPAC) and CGRP, respectively.

Article Snippet: Calcitonin gene-related peptide 8-37 (CGRP 8-37 ), capsaicin, glibenclamide, P-(4-methoxyphenyl)-P-4-morpholinylphosphinedithioic acid (GYY4137), 2-(1,3-dimethyl-2,6-dioxo-1,2,3,6-tetrahydro-7H-purin-7-yl)-N-(4-isopropyl phenyl)acetamide (HC030031), (9R,10S,12S)-2,3,9,10,11,12-hexahydro-10-hydroxy-9-methyl-1-oxo-9,12-epoxy-1H-diindolo[1,2,3-fg:3',2',1'-kl]pyrrolo[3,4-] , benzodiazocine -10-carboxylic acid (KT5720), 1H- , , -oxadiazolo[4,3-a]quinoxalin-1-one (ODQ), pituitary adenylyl cyclase-activating polypeptide 6-38 (PACAP 6-38 ) and 9,11-dideoxy-9a,11a-methanoepoxy prostaglandin F 2α (U46619) from Tocris (Bristol, UK).

Techniques: Membrane, Activation Assay

Journal: The Journal of Headache and Pain

Article Title: PACAP6-38 improves nitroglycerin-induced central sensitization by modulating synaptic plasticity at the trigeminal nucleus caudalis in a male rat model of chronic migraine

doi: 10.1186/s10194-023-01603-3

Figure Lengend Snippet: PAC1R inhibitor PACAP6-38 alleviated central sensitization and reduced neuronal activation in the TNC of CM rat model. A - C . PACAP6-38 treatment ameliorated the decreased mechanical ( A - B ) and thermal ( C ) thresholds after CM. Mean ± SEM, n = 6/group. Two-way ANOVA with the Bonferroni post hoc test, * p < 0.05, ** p < 0.01, *** p < 0.001 compared with saline + vehicle group, & p < 0.05, && p < 0.01, &&& p < 0.001 compared with NTG + vehicle group. D . Representative western blot bands and densitometric quantification of c-Fos in different groups. E – F . Representative micrographs ( E ) and quantitative analysis ( F ) of c-Fos (red) staining in TNC. The nuclei were stained with DAPI (blue). Mean ± SEM, n = 3/group. One-way ANOVA, Dunnett; * p < 0.05, ** p < 0.01, *** p < 0.001 compared with saline + vehicle group, & p < 0.05, && p < 0.01, &&& p < 0.001 compared with NTG + vehicle group. Abbreviations: PA6-38, pituitary adenylate cyclase-activating peptide 6–38; NTG, nitroglycerin; CM: chronic migraine

Article Snippet: PACAP6-38 (HY-P0220A, MCE) was dissolved in 0.9% saline to make a working solution of 1 μg/μl.

Techniques: Activation Assay, Saline, Western Blot, Staining

Journal: The Journal of Headache and Pain

Article Title: PACAP6-38 improves nitroglycerin-induced central sensitization by modulating synaptic plasticity at the trigeminal nucleus caudalis in a male rat model of chronic migraine

doi: 10.1186/s10194-023-01603-3

Figure Lengend Snippet: PACAP6-38 administration decreased the overexpression of synaptic-associated proteins and restored the aberrant synaptic ultrastructure in CM rat model. A - C . Representative bands and quantification of PSD-95, syt-1 and syp protein levels showed that these proteins were significantly increased in the NTG + vehicle group compared to the saline + vehicle group, while PACAP6-38 blocked these effects. E – G . Representative images of the synaptic ultrastructure in the four groups. H - J . The width of the synaptic cleft ( H ) was decreased, and the thickness of PSD ( I ) and the synaptic interface curvature ( J ) were significantly increased in the NTG + VEHgroup compared to the Saline + VEHgroup. In the NTG + PA6-38 group, these abnormal alterations were alleviated. Mean ± SEM. n = 3/group. One-way ANOVA with Dunnett’s post hoc test; Scale bars = 500 nm. * p < 0.05, ** p < 0.01, *** p < 0.001 compared with saline + vehicle group, & p < 0.05, && p < 0.01, &&& p < 0.001 compared with NTG + vehicle group. Abbreviations: PA6-38, pituitary adenylate cyclase-activating peptide 6–38; NTG, nitroglycerin; CM: chronic migraine

Article Snippet: PACAP6-38 (HY-P0220A, MCE) was dissolved in 0.9% saline to make a working solution of 1 μg/μl.

Techniques: Over Expression, Saline

Journal: The Journal of Headache and Pain

Article Title: PACAP6-38 improves nitroglycerin-induced central sensitization by modulating synaptic plasticity at the trigeminal nucleus caudalis in a male rat model of chronic migraine

doi: 10.1186/s10194-023-01603-3

Figure Lengend Snippet: Inhibiting PAC1R with PACAP6-38 restored the aberrant neuronal dendritic spines via the ERK/CREB/BDNF signaling pathway in the CM rat model. A - D . Representative images of the dendritic spines in different groups. E . Golgi-Cox staining showed that the number of dendritic spines per 20 μm was significantly higher in the NTG + VEH group compared to the Saline + VEH group, and the effect was inhibited in the NTG + PA6-38 group. (F–H). Representative bands showed that the protein levels of p-ERK( F ), p-CREB ( J ) and BDNF ( H ) were significantly elevated in the NTG + VEH group compared to the Saline + VEH group. After PACAP6-38 treatment, the expression levels of these proteins were significantly reduced. Mean ± SEM, n = 3/group. One-way ANOVA with Dunnett’s post hoc test; * p < 0.05, ** p < 0.01, *** p < 0.001 compared with saline + vehicle group, & p < 0.05, && p < 0.01, &&& p < 0.001 compared with NTG + vehicle group. Abbreviations: PA6-38, pituitary adenylate cyclase-activating peptide 6–38; NTG, nitroglycerin; CM: chronic migraine

Article Snippet: PACAP6-38 (HY-P0220A, MCE) was dissolved in 0.9% saline to make a working solution of 1 μg/μl.

Techniques: Staining, Saline, Expressing

Journal: British Journal of Pharmacology

Article Title: Activation of PAC 1 and VPAC receptor subtypes elicits differential physiological responses from sympathetic preganglionic neurons in the anaesthetized rat

doi: 10.1111/j.1476-5381.2012.02045.x

Figure Lengend Snippet: In vivo effects of intrathecal VPAC1 receptor activation with 15 min of PACAP(6–38) followed by PACAP on MAP, HR and sSNA. (A) Experimental record of VPAC1 receptor activation following intrathecal PACAP(6–38) followed by PACAP, on (i) AP (MAP is represented by the black line on the AP trace), (ii) HR and (iii) sSNA over 15 min and 1 h recording periods, respectively. Arrows indicate times of 1000 µmol·L−1 PACAP(6–38) and 1000 µmol·L−1 PACAP administration. Note the lack of effect of PACAP(6–38). (B) Changes in (i) MAP, (ii) HR and (iii) sSNA after PACAP(6–38) only (n= 6) and after VPAC1 receptor activation (n= 6). PACAP(6–38) alone does not affect MAP, HR and sSNA compared with responses to vehicle (PBS). PACAP(6–38) followed by PACAP (VPAC1 activation) decreased MAP (Δ-20 ± 7 mmHg; n= 6), and increased both HR (Δ 17 ± 4 bpm; n= 6) and sSNA (Δ 32 ± 14%; n= 6). Arrow indicates time of PACAP(6–38) or PACAP injection. ‘PBS’ is the period after intrathecal infusion of vehicle (PBS). ‘Drug’ is the period after intrathecal injection of PACAP(6–38) only or PACAP injection following PACAP(6–38). *P < 0.05, **P < 0.01, ***P < 0.0001, significantly different from PBS.

Article Snippet: Rats were pretreated with 1000 µmol·L −1 PACAP(6–38) (Auspep), a competitive PAC 1 and VPAC 2 receptor antagonist (K d = 1.5 nM) ( Robberecht et al ., 1992 ; Vaudry et al ., 2009 ; Farnham et al ., 2011 ), for 15 min ( n = 6) and 30 min ( n = 4) prior to PACAP-38 (1000 µmol·L −1 ; Auspep) administration.

Techniques: In Vivo, Activation Assay, Injection