Pituitary Adenylate Cyclase-Activating Polypeptide Expression and Modulation of Neuronal Excitability in Guinea Pig Cardiac Ganglia

Braas, Karen M.; May, Víctor; Harakall, Susan A.; Hardwick, Jean C.; Parsons, Rodney L.

doi:10.1523/jneurosci.18-23-09766.1998

Cited by 103 publications

(164 citation statements)

References 69 publications

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Section: Methodsmentioning

confidence: 99%

“…Cardiac ganglia preparations were prepared as described previously (Mawe et al, 1996;Braas et al, 1998;Calupca et al, 2000). The preparations were maintained in Sylgard-lined dishes with oxygenated Krebs solution at 35 -37°C.…”

Section: Methodsmentioning

confidence: 99%

“…The cardiac ganglia preparations were immunolabeled using procedures described previously (Mawe et al, 1996;Braas et al, 1998;Calupca et al, 2000;Parsons et al, 2006;Girard et al, 2006a,b). The fixed atrial tissue was washed in phosphate buffered saline, permeabilized with 0.5% Triton X-100 and incubated at 4°C overnight with primary antiserum (mouse monoclonal anti-PACAP 1:10, from Dr. Jan Fahrenkrug, Copenhagen, Denmark; rabbit anti-VIP 1:100 from CURE Research Center, UCLA, Los Angeles, CA; goat anti-choline acetyl transferase (ChAT) 1:100, obtained from Chemicon International, Temecula, CA; and rabbit anti-PGP 9.5 1:1000 from Ultraclomne Ltd, Isle of Wright, UK).…”

Section: Immunohistochemistrymentioning

confidence: 99%

“…The number of cardiac neurons in each preparation was determined by counting ChAT-IR or PGP-9.5-IR cells (Mawe et al, 1996;Braas et al, 1998;Kennedy et al, 1998;Lynch et al, 1999;Calupca et al, 2000;Braas et al, 2004,Parsons et al, 2006. PGP-95-IR neurons) X 100 = %].…”

Section: Quantification Of Peptide-ir Cardiac Neuronsmentioning

confidence: 99%

“…The preparations were maintained in Sylgard-lined dishes with oxygenated Krebs solution at 35 -37°C. When prepared for immunocytochemistry, control and cultured preparations were fixed in 2% paraformaldehyde containing 0.2% picric acid for 2 hours at room temperature (Mawe et al, 1996;Braas et al, 1998;Lynch et al, 1999;Calupca et al, 2000;Parsons et al, 2006).For culture, the cardiac ganglia explants were dissected under sterile conditions and then maintained at 37°C in culture media consisting of DMEM-F12 (1:1) containing 10% horse serum, gentamicin (10 μg/ml), amphotericin B (3.75 μg/ml), penicillin (100 units/ml) and streptomycin (100 μg/ml) (Sigma Chemical Company, St. Louis, MO, USA) (Lynch et al, 1999;Calupca et al, 2000;Braas et al, 2004;Girard et al, 2006a …”

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Regulation of neuronal pituitary adenylate cyclase-activating polypeptide expression during culture of guinea-pig cardiac ganglia

et al. 2007

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The trophic neuropeptide pituitary adenylate cyclase-activating polypeptide (PACAP) increases in many different neuron types following injury; a response postulated to support cell survival and regeneration. In acutely isolated cardiac ganglia, approximately 1% of the cardiac neurons exhibited PACAP immunoreactivity whereas after 72 hours in culture, ~25% of the neurons were PACAP immunoreactive. In contrast, there was no increase in vasoactive intestinal polypeptide (VIP)-immunoreactive (IR) cells. Using a combination of immunocytochemical and molecular techniques, we have quantified PACAP expression, during explant culture of guinea pig cardiac ganglia. Using real time PCR, PACAP transcript levels increased progressively up to 48 hours in culture with no further increase after 72 hours. PACAP transcript levels were reduced by neurturin at 48 hours in culture but not after 24 or 72 hours in culture. In addition, neurturin partially suppressed the percentage of PACAP-IR neurons after 72 hours in culture, an effect mediated by activation of the phosphatidylinositol 3-kinase and mitogen activated protein kinase signaling pathways. The addition of different known regulatory molecules, including CNTF, Il-1β, TNFα, bFGF, TGF βand NGF did not increase the percentage of PACAP-IR neurons after 24 hours in culture; a result indicating that the generation and secretion of these factors did not stimulate PACAP expression. The presence of 20 nM PACAP or 10 μM forskolin increased the percentage of PACAP-IR cardiac neurons in 24 hour cultures, but not in 72 hour cultures. Neither treatment enhanced the number of VIP-IR neurons. The addition of the PAC 1 receptor antagonist, M65 (100 nM) suppressed the 20 nM PACAP-induced increase in percentage of PACAP-IR cells in 24 hour cultures indicating the effect of PACAP was mediated through the PAC 1 receptor. However, 100 nM M65 had no effect on the percentage of PACAP-IR cells in either 24 or 48 hour cultures not treated with exogenous PACAP, suggesting that endogenous release of PACAP likely did not contribute to the enhanced peptide expression. We postulate that the enhanced PACAP expression, which occurs in response to injury is facilitated in the explant cultured cardiac ganglia by the loss of a target-derived inhibitory factor, very likely neurturin. In intact tissues the presence of neurturin would normally suppress PACAP expression. Lastly, our results indicate that many common trophic factors do not enhance PACAP expression in the cultured cardiac neurons. However, the stimulatory role of an, as yet, unidentified factor can not be excluded. Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content...

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Immunohistochemistrymentioning

confidence: 99%

Section: Quantification Of Peptide-ir Cardiac Neuronsmentioning

confidence: 99%

mentioning

confidence: 99%

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Regulation of neuronal pituitary adenylate cyclase-activating polypeptide expression during culture of guinea-pig cardiac ganglia

et al. 2007

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Origin of pituitary adenylate cyclase-activating polypeptide (PACAP)-immunoreactive fibers innervating guinea pig parasympathetic cardiac ganglia

2000

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The present study investigated the origin of pituitary adenylate cyclase-activating polypeptide (PACAP) -immunoreactive (IR) fibers innervating guinea pig cardiac ganglia. Immunohistochemistry was performed on whole-mounts containing cardiac ganglia, and sections of stellate, nodose, and dorsal root ganglia (DRG, thoracic levels 1-4), and caudal medulla. In control preparations, only 4% of the cardiac neurons were PACAP-IR, although most cardiac ganglion cells were surrounded by a network of PACAP-IR fibers. After 3-7 days in explant culture, the number of PACAP-IR cardiac neurons increased approximately eightfold. However, virtually all PACAP-IR fibers surrounding the cardiac neurons had degenerated, demonstrating that the major source of the PACAP-IR fibers was extrinsic to the cardiac ganglia preparation. PACAP- and choline acetyltransferase (ChAT) immunoreactivity were colocalized in fibers within the stellate ganglia but not within neuropeptide Y (NPY) -IR cell bodies and fibers. PACAP-IR cells and fibers were present in the nodose ganglia. PACAP immunoreactivity also was present in fibers and primarily small neurons in thoracic DRGs. In situ hybridization demonstrated the presence of proPACAP mRNA within neurons in the region of the dorsal motor nucleus of the vagus and nucleus ambiguus. PACAP immunoreactivity was colocalized with ChAT immunoreactivity, but not with NPY immunoreactivity or SP immunoreactivity, in fibers surrounding neurons within cardiac ganglia. We conclude that PACAP-containing fibers innervating the postganglionic parasympathetic neurons in guinea pig cardiac ganglia are primarily preganglionic parasympathetic axons.

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Pituitary adenylate cyclase activating peptide (PACAP) in the enteropancreatic innervation

Kirchgessner

Liu

2001

Anat. Rec.

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Pituitary Adenylate Cyclase-Activating Polypeptide Expression and Modulation of Neuronal Excitability in Guinea Pig Cardiac Ganglia

Cited by 103 publications

References 69 publications

Regulation of neuronal pituitary adenylate cyclase-activating polypeptide expression during culture of guinea-pig cardiac ganglia

Regulation of neuronal pituitary adenylate cyclase-activating polypeptide expression during culture of guinea-pig cardiac ganglia

Origin of pituitary adenylate cyclase-activating polypeptide (PACAP)-immunoreactive fibers innervating guinea pig parasympathetic cardiac ganglia

Pituitary adenylate cyclase activating peptide (PACAP) in the enteropancreatic innervation

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