Journal: Neural Regeneration Research

Article Title: Inhibition of retinal ganglion cell apoptosis: regulation of mitochondrial function by PACAP

doi: 10.4103/1673-5374.232489

Figure Lengend Snippet: Densitometric analysis of PAC1 and Bcl-2 levels in RGC-5 cells exposed to SD with or without 100 nM PACAP. SD caused a significant decrease in the level (optical value ratio) of Bcl-2 (B) and a concomitant increase of PAC1 (A), which was counteracted by PACAP treatment. # P < 0.05, ## P < 0.05, vs . SD group (mean ± SD, one-way analysis of variance followed by the Student-Newman-Keuls test). Each experiment was conducted three times. PACAP: Pituitary adenylate cyclase-activating polypeptide; RGC: retinal ganglion cell; SD: serum deprivation.

Article Snippet: Membranes were incubated at 4°C with primary antibodies as follows: mouse monoclonal anti-Bcl-2 antibody (1:1000; Santa Cruz Biotechnology, Santa Cruz, CA, USA), rabbit polyclonal anti-PAC1 antibody (1:1000; Santa Cruz Biotech nology), and mouse monoclonal anti-β-actin antibody (1:3000; Cell Signaling, Danvers, MA, USA).

Techniques:

Journal: Journal of Endocrinology

Article Title: Pituitary adenylate cyclase-activating polypeptide, vasoactive intestinal polypeptide and their receptors: distribution and involvement in the secretion of Podarcis sicula adrenal gland

doi: 10.1677/joe-07-0127

Figure Lengend Snippet: Figure 5 Effects of VIP 10K7 M administration on co-cultured adrenal cells. Data are expressed in pg/mlGS.E.M. PACAP 6–38 (10K6 M) was added, where appropriate, to avoid endogenous PACAP stimulation. (a) Time course of catecholamine after VIP ( ) or VIPCVPAC1-A (10K6 M) (B) administration. P!0.01 versus baseline. (b) Time course of corticosterone (squares) and aldosterone (triangles) after VIP (black) or VIPCVPAC1-A (empty) administration. *P!0.05; **P!0.01; ***P!0.001 versus baseline. **P!0.01 when VIP is compared with VIP plus antagonist for aldosterone. (c) VIP effects on catecholamine and steroids through VPAC2 receptor exclusively. To block simultaneously VPAC1, b-adrenoceptors and PAC1, samples were treated with VIPCVPAC1-A (10K6 M)CPropanolol (10K6 M)CPACAP 6–38 (10K6 M) respectively. *P!0.05. (d) VIP effects on catecholamine and steroids through VPAC1 receptor exclusively. To block PAC1, b-adrenoceptors and VPAC2 pathways, VPAC2 immunoneutralized adrenals were treated with VIPCPropanolol (10K6 M)CPACAP 6–38 (10K6 M). *P!0.05.

Article Snippet: Overnight incubation was performed at 4 8C with a rabbit anti-PACAP27 antibody (Phoenix Pharmaceuticals) diluted 1:300 in normal goat serum or anti-VPAC1, anti-VPAC2 and anti-PAC1 antibody (Santacruz Biotechnology, Santa Cruz, CA, USA) diluted 1:100.

Techniques: Cell Culture, Blocking Assay

Journal: Journal of Endocrinology

Article Title: Pituitary adenylate cyclase-activating polypeptide, vasoactive intestinal polypeptide and their receptors: distribution and involvement in the secretion of Podarcis sicula adrenal gland

doi: 10.1677/joe-07-0127

Figure Lengend Snippet: Figure 7 Effect of PACAP 10K7 M administration on co-cultured adrenal cells. Data expressed in pg/mlGS.E.M. (a) Catecholamine secretion after PACAP ( ) and PACAPCVPAC1-A 10K6 M (B). P!0.05 versus baseline. (b) Corticosterone (squares) and aldosterone (triangles) secretion after PACAP (black) and PACAPC VPAC1-A 10K6 M (empty) administration. **P!0.01; ***P!0.001 versus baseline. P!0.01 when PACAP is compared with PACAP plus antagonist for aldosterone and corticosterone. (c) PACAP effects through VPAC1 and VPAC2 receptors on catecholamine and steroids secretion. Samples were treated with Propanolol (10K6 M)CPACAP 6–38(10K6 M). *P!0.05. (d) Effects of PACAPadministration on catecholamine and steroids through PAC1 receptor exclusively. To block simultaneously VPAC2, b-adrenoceptors and VPAC1, VPAC2 immunoneutralized adrenal cells were treated with PACAPCPropanolol (10K6 M)CVPAC1-A (10K6 M). *P!0.05. (e) PACAP effects on catecholamine and steroids through VPAC2 receptor exclusively. To block simultaneously b-adrenoceptors, VPAC1 and PAC1, samples were treated with PACAPCPropanolol (10K6 M)CVPAC1-A (10K6 M)CPACAP 6–38(10K6 M). *P!0.05. (f) PACAP effects on catecholamine and steroids through VPAC1 receptor exclusively. To block simultaneously VPAC2, b-adrenoceptors and PAC1, VPAC2 immunoneu- tralized adrenal cells were treated with PACAPCPropanolol (10K6 M)CPACAP 6–38 (10K6 M) respectively. *P!0.05.

Article Snippet: Overnight incubation was performed at 4 8C with a rabbit anti-PACAP27 antibody (Phoenix Pharmaceuticals) diluted 1:300 in normal goat serum or anti-VPAC1, anti-VPAC2 and anti-PAC1 antibody (Santacruz Biotechnology, Santa Cruz, CA, USA) diluted 1:100.

Techniques: Cell Culture, Blocking Assay

Journal: REPRODUCTION

Article Title: Expression and in vitro regulation of pituitary adenylate cyclase-activating polypeptide (pacap38) and its type I receptor (pac1-r) in the gonads of tilapia (Oreochromis mossambicus)

doi: 10.1530/rep-08-0422

Figure Lengend Snippet: Figure 1 Semi-quantitative RT-PCR analysis of male (A and C) and female (B and D) tilapia tissues using specific primers for tpacap38 (A and B) and pacap type I receptor (tpac1-r; C and D). (Representative pictures are shown). B, brain; Ga, gallbladder; Gi, gill; H, heart; I, intestine; K, kidney; L, liver; Mu, muscle; P, pancreas; Sp, spleen; St, stomach; O, ovary; T, testis. a,b,c,d,e,fDifferent superscripts among groups indicate statistically significant differences (P!0.05).

Article Snippet: Sections were pretreated with 3% H2O2 (100% methanol and 3% H2O2 mixed 1:1) in methanol for inactivation of endogenous peroxidase, and the sections were incubated with anti-PACAP and PACAP type I receptor (PAC1-R) polyclonal antibodies (0.2 mg in 50 ml PBS, sc-25439 and sc30018, rabbit anti human origin; Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA) for 1 h at RTor overnight at 4 8C and then washed three times in PBS.

Techniques: Quantitative RT-PCR

Journal: REPRODUCTION

Article Title: Expression and in vitro regulation of pituitary adenylate cyclase-activating polypeptide (pacap38) and its type I receptor (pac1-r) in the gonads of tilapia (Oreochromis mossambicus)

doi: 10.1530/rep-08-0422

Figure Lengend Snippet: Figure 2 Immunohistochemical staining of PACAP and PACAP type I receptor in brain (A), gallbladder (B), gill (C), heart (D), intestine (E), kidney (F), liver (G), muscle (H), pancreas (I), spleen (J), stomach (K), ovary (L) testis (M), and 1-month-old whole juveniles of tilapia. Control groups of PACAP are shown. The blue arrowheads indicate PACAPand PACAP type I receptor proteins. BrC, brain cortex; BrW, brain white matter; CaC, centroacinar cells; CaM, cardic muscle; Du, duct; GiE, epithelial of gill; Gra, granulosa cells; Hep, hepatocytes; InE,epithelial of intestines; InC, interstitial cells; LP, lamina propira; MuF, muscular filament; O, oocyte; PT, portal tract; RP, red pulp; RT, renal tubules; Sp, sperm; The, theca cells; SM, submucosa cells; StE, epithelial of stomach. A2–M2 and A4–M4 are the insets from A1–M1 and A3–M3 respectively. A–M, A1–M1 and A3–M3: Scale barZ100 mm; A2–M2 and A4–M4: Scale barZ20 mm; N and O: Scale barZ1 mm; N1, N2, O1 and O2: Scale barZ20 mm.

Article Snippet: Sections were pretreated with 3% H2O2 (100% methanol and 3% H2O2 mixed 1:1) in methanol for inactivation of endogenous peroxidase, and the sections were incubated with anti-PACAP and PACAP type I receptor (PAC1-R) polyclonal antibodies (0.2 mg in 50 ml PBS, sc-25439 and sc30018, rabbit anti human origin; Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA) for 1 h at RTor overnight at 4 8C and then washed three times in PBS.

Techniques: Immunohistochemical staining, Staining, Control

Journal: eLife

Article Title: Light-at-night exposure affects brain development through pineal allopregnanolone-dependent mechanisms

doi: 10.7554/eLife.45306

Figure Lengend Snippet: ( a ) Shown are the effects of Px or Px plus ALLO on the expression of neuroprotective and neurotoxic factors ( n = 10). ( b ) Diurnal changes in PACAP mRNA expression in the cerebellum under LD or light-at-night conditions in P7 chicks ( n = 10). ( c ) Number of Purkinje cells expressing active caspase-3 in lobule IV at P7 ( n = 10). ( d ) Number of Purkinje cells in lobule IV at P21 ( n = 10). Immunohistochemistry of PACAP ( e ) and PAC1 ( f ) in the cerebellum of P1 chicks (similar results were obtained in repeated experiments using three different samples). Calbindin is used as a marker for Purkinje cells. ML, molecular layer; PCL, Purkinje cell layer; GCL, granule cell layer; LAN, light-at-night. 10.7554/eLife.45306.018 Figure 5—source data 1. source data for mRNA expressions ( a ), diurnal changes in PACAP mRNA expression ( b ), and numbers of Purkinje cells ( c and d ) in male.

Article Snippet: After blocking nonspecific binding with Protein Block, serum-free (Agilent Technologies, Palo Alto, CA), the sections were immersed overnight at 4°C in either a 1:100 dilution of rabbit anti-goldfish mPRα antiserum, a 1:1000 dilution of chicken anti-PACAP antibody , or a 1:100 dilution of mouse anti-PAC1 antibody (sc-100315; Santa Cruz Biotechnology, Santa Cruz, CA).

Techniques: Expressing, Immunohistochemistry, Marker

Journal: eLife

Article Title: Light-at-night exposure affects brain development through pineal allopregnanolone-dependent mechanisms

doi: 10.7554/eLife.45306

Figure Lengend Snippet:

Article Snippet: After blocking nonspecific binding with Protein Block, serum-free (Agilent Technologies, Palo Alto, CA), the sections were immersed overnight at 4°C in either a 1:100 dilution of rabbit anti-goldfish mPRα antiserum, a 1:1000 dilution of chicken anti-PACAP antibody , or a 1:100 dilution of mouse anti-PAC1 antibody (sc-100315; Santa Cruz Biotechnology, Santa Cruz, CA).

Techniques: Recombinant, Membrane, Sonication, Chromatin Immunoprecipitation, Software