pacap(1–27) peptide  (GL Biochem)

Journal: International Journal of Molecular Sciences

Article Title: Polarized Anti-Inflammatory Mesenchymal Stem Cells Increase Hippocampal Neurogenesis and Improve Cognitive Function in Aged Mice

doi: 10.3390/ijms24054490

Figure Lengend Snippet: Polarized PACAP-treated MSC maintain mesenchymal phenotype. ( A ). Immunophenotyping of PACAP-treated MSC in flow cytometry presents positive expression of the mesenchymal markers CD106 (>50%), CD29 (>80%), CD44 (>70%), CD73 (>60%) and sca-1 (>40%) but negative expression of the hematopoietic markers CD45 (<8%), and CD11b (<5%). Graphs represent flow cytometry histograms for the expression of the different markers. The negative control histogram is presented with the blue filled histogram. ( B ). Both naïve (N = 4) and PACAP-treated MSC (N = 3) expressed detectable levels of VPAC2 receptor mRNA, as detected in real-time PCR. Since the activation of Toll-like receptor 3 (TLR3) is an established marker of the MSC anti-inflammatory phenotype (MSC2) , we administered pituitary adenylate cyclase-activating peptide (PACAP), a neuropeptide with anti-inflammatory properties that is known to upregulate TLR3 and vice versa with TLR4, at 20 nM for 4 days to establish the anti-inflammatory MCS phenotype (MSC2). PACAP treatment of MSC in vitro (pMSC, N = 5) did not increase significantly the expression of TLR3 ( C ) or TLR4 ( D ) but increased the TLR3/TLR4 gene expression ratio ( E ) compared with naïve MSC (N = 4), as detected in real-time PCR. All graphs present mean ± SE. * p < 0.05, Student t -test. ns = non-significant.

Article Snippet: Pituitary adenylate cyclase-activating peptide (PACAP) amino acids 1–27 was purchased from Bachem AG (Switzerland) cat. No. 127317-03-7.

Techniques: Flow Cytometry, Expressing, Negative Control, Real-time Polymerase Chain Reaction, Activation Assay, Marker, In Vitro, Gene Expression

Journal:

Article Title: The P 2 -purinoceptor antagonist suramin is a competitive antagonist at vasoactive intestinal peptide receptors in the rat gastric fundus

doi: 10.1038/sj.bjp.0703482

Figure Lengend Snippet: (a) Magnitude of relaxant responses to pituitary adenylate cyclase activating peptide 1–27 (PACAP; 0.1–100 nM) in longitudinal strips of rat gastric fundus in the absence and presence of α-chymotrypsin (1 u ml−1) or suramin (3–200 μM). (b) Schild plot analysis of the antagonism by suramin of relaxant responses to PACAP, where CR is the concentration ratio for PACAP required to produce the same response in the presence and absence of suramin. The analysis indicated that suramin had a pA2 value of 5.6±0.1 (slope=1.0±0.1). Values are mean±s.e.mean for 4–6 experiments.

Article Snippet: The following drugs were used in the study: adenosine 5′-triphosphate disodium salt (ATP, Sigma, U.S.A.), atropine sulphate (Sigma, U.S.A.), α-chymotrypsin (bovine pancreas, Sigma, U.S.A.), guanethidine sulphate (Ciba-Geigy, Australia), 5-hydroxytryptamine creatinine sulphate (serotonin; Sigma, U.S.A.), isoproterenol hydrochloride (isoprenaline, Sigma, U.S.A.), nitric oxide gas (NO; CIG, Australia), pituitary adenylate cyclase activating peptide 1–27 (PACAP, ovine, Auspep, Australia), sodium nitroprusside (SNP; Sigma, U.S.A.), tetrodotoxin (Sigma, U.S.A.), vasoactive intestinal peptide (VIP, human; Auspep, Australia).

Techniques: Concentration Assay