Involvement Of Pituitary Adenylate Cyclase-activating Polypeptide Research Articles

Differential Vulnerability of Oculomotor Versus Hypoglossal Nucleus During ALS: Involvement of PACAP.

Amyotrophic lateral sclerosis (ALS) is a progressive multifactorial disease characterized by the loss of motor neurons (MNs). Not all MNs undergo degeneration: neurons of the oculomotor nucleus, which regulate eye movements, are less vulnerable compared to hypoglossal nucleus MNs. Several molecular studies have been performed to understand the different vulnerability of these MNs. By analyzing postmortem samples from ALS patients to other unrelated decedents, the differential genomic pattern between the two nuclei has been profiled. Among identified genes, adenylate cyclase activating polypeptide 1 (ADCYAP1) gene, encoding for pituitary adenylate cyclase-activating polypeptide (PACAP), was found significantly up-regulated in the oculomotor versus hypoglossal nucleus suggesting that it could play a trophic effect on MNs in ALS. In the present review, some aspects regarding the different vulnerability of oculomotor and hypoglossal nucleus to degeneration will be summarized. The distribution and potential role of PACAP on these MNs as studied largely in an animal model of ALS compared to controls, will be discussed.

Pituitary adenylate cyclase-activating polypeptide (PACAP) contributes to the proliferation of hematopoietic progenitor cells in murine bone marrow via PACAP-specific receptor.

Pituitary adenylate cyclase-activating polypeptide (PACAP, encoded by adcyap1) plays an important role in ectodermal development. However, the involvement of PACAP in the development of other germ layers is still unclear. This study assessed the expression of a PACAP-specific receptor (PAC1) gene and protein in mouse bone marrow (BM). Cells strongly expressing PAC1+ were large in size, had oval nuclei, and merged with CD34+ cells, suggesting that the former were hematopoietic progenitor cells (HPCs). Compared with wild-type mice, adcyap1−/− mice exhibited lower multiple potential progenitor cell populations and cell frequency in the S-phase of the cell cycle. Exogenous PACAP38 significantly increased the numbers of colony forming unit-granulocyte/macrophage progenitor cells (CFU-GM) with two peaks in semi-solid culture. PACAP also increased the expression of cyclinD1 and Ki67 mRNAs. These increases were completely and partially inhibited by the PACAP receptor antagonists, PACAP6-38 and VIP6-28, respectively. Little or no adcyap1 was expressed in BM and the number of CFU-GM colonies was similar in adcyap1−/− and wild-type mice. However, PACAP mRNA and protein were expressed in paravertebral sympathetic ganglia, which innervate tibial BM, and in the sympathetic fibers of BM cavity. These results suggested that sympathetic nerve innervation may be responsible for PACAP-regulated hematopoiesis in BM, mainly via PAC1.

Gene Interference with Morpholinos in a Gold Nanoparticle-Based Delivery Platform in Rat PC12 Cells.

For the first time the efficiency of gene knockdown of the pituitary adenylate cyclase-activating polypeptide (PACAP) receptor 1 (PAC1R) is demonstrated by employing gold nanocomplexes. This gene knockdown subsequently affects the action of PACAP on neurite outgrowth in PC12 cells. These nanocomplexes comprise cholera toxin B (CTB)-gold nanoparticle conjugates loaded with double-stranded morpholinos (MOs) (photo MO and antisense MO). Nanocomplexes are introduced into cells via lipid raft-dependent endocytosis. After UV light exposure, the photolinker in the photo MO is cleaved, bisecting the photo MO and releasing the antisense MO from the conjugate. The antisense MO then binds the PAC1R mRNA and decreases gene expression. The maximal efficiency of gene knockdown is observed after 24 hours, resulting in a 65% ± 12 reduction of the protein level. This reduction in PAC1R impairs the responsiveness of cells to PACAP exposure. Following PAC1R gene knockdown, only 10% ± 8 and 11%± 9 of cells exhibit neurite outgrowth after 4-day exposure to PACAP-38 and PACAP-27, respectively. These results demonstrate an efficient PAC1R gene knockdown and noticeable difference in response to PACAP action on neural cell differentiation, adding an extra dimension to determine the involvement of PACAP and its PAC1R in the neurotropic effect to PC12 cells.

Pituitary adenylate cyclase-activating polypeptide (PACAP) stimulates release of somatolactin (SL)-α and SL-β from cultured goldfish pituitary cells via the PAC1 receptor-signaling pathway, and affects the expression of SL-α and SL-β mRNAs

Pituitary adenylate cyclase-activating polypeptide (PACAP) is a neuropeptide that stimulates the release of adenohypophyseal hormone from the pituitary in fish. In the goldfish, PACAP induces the release of somatolactin (SL), in particular, from cultured pituitary cells. SL belongs to the growth hormone and prolactin family, and comprises two molecular variants termed SL-α and SL-β in goldfish. However, there is no information about the involvement of PACAP in the regulation of SL-α and SL-β release and the expression of their mRNAs. Therefore, we examined the effect of PACAP on SL-α and SL-β release from cultured goldfish pituitary cells. Treatment with PACAP (10−10–10−7M) increased the release of both SL-α and SL-β. The stimulatory action of PACAP (10−9M) on SL-α and SL-β release was blocked by treatment with a PACAP-selective receptor (PAC1R) antagonist, PACAP(6–38) (10−6M). We also examined whether PACAP affects the expression of SL-α and SL-β mRNAs in cultured pituitary cells. Treatment with PACAP (10−9 and 10−8M) for 6h decreased the expression level of SL-α mRNA but increased that of SL-β mRNA. The action of PACAP (10−8M) on SL-β mRNA expression was blocked by treatment with PACAP(6–38) (10−6M), whereas PACAP(6–38) elicited no change in the expression of SL-α mRNA. These results indicate that in cultured goldfish pituitary cells, PACAP stimulates the release of SL-α and SL-β, and expression of SL-β mRNA, via the PAC1R-signaling pathway. However, the mechanism whereby PACAP inhibits the expression of SL-α mRNA does not seem to be mediated by PAC1R signaling.

Possible Role of PACAP and Its PAC1 Receptor in the Differential Regulation of Pituitary LHbeta- and FSHbeta-Subunit Gene Expression by Pulsatile GnRH Stimulation1

The pituitary gonadotropins, luteinizing hormone (LH) and follicle-stimulating hormone (FSH), are mainly under the control of hypothalamic gonadotropin-releasing hormone (GnRH), which regulates male and female gonadal function. GnRH is released in a pulsatile manner from the hypothalamus, and the frequency of GnRH pulses determines the dominance of output of LH and FSH from pituitary gonadotrophs. That is, more rapid pulses of GnRH preferentially increase synthesis and secretion of LH, whereas FSH is preferentially stimulated by slower GnRH pulses. The detailed mechanisms underlying this phenomenon remain unknown. Pituitary adenylate cyclase-activating polypeptide (PACAP) was originally identified as a hypothalamic activator of cAMP production in pituitary cells. PACAP is produced within the pituitary gonadotroph as well as in the central nervous system. PACAP stimulates gonadotropin alpha-, LHbeta-, and FSHbeta-subunits as well as receptors for GnRH in the pituitary gonadotropin-secreting cells. In addition, its own receptor, PACAP type I receptor (PAC1R), is also regulated by PACAP in gonadotrophs. GnRH stimulates expression of PACAP as well as PAC1R, and lower frequencies of GnRH pulses preferentially increase PACAP and PAC1R expression in gonadotrophs. Increasing concentrations of PACAP further increase the levels of gonadotropin subunit and that increasing amounts of PAC1R in gonadotrophs potentiates the effects of PACAP or GnRH on gonadotropin subunit expression. In addition, we have observed that GnRH-increased FSHbeta-subunit expression was prevented in the presence of PAC1R antagonist. These observations suggest the involvement of locally produced PACAP and its PAC1R in the differential regulation of specific gonadotropin subunit expression by pulsatile GnRH stimulation. Here, we review the possible involvement of PACAP and its PAC1R in gonadotropin control on the basis of our observations with gonadotroph cell lines.

Comparative Examination of Inner Ear in Wild Type and Pituitary Adenylate Cyclase Activating Polypeptide (PACAP)-Deficient Mice

Pituitary adenylate cyclase activating polypeptide (PACAP) is a multifunctional neuropeptide with well-known neuroprotective and neurotrophic effects. The involvement of PACAP in sensory processing has also been documented, but little is known about its effects in the auditory system. PACAP and its specific receptor (PAC1) are present in the cochlea and in brain structures involved in auditory pathways. Recently, we have shown that PACAP protects cochlear cells against oxidative stress-induced apoptosis. The endolymphatic Ca(2+) concentration controlled by Ca(2+) buffers of the hair cells is essential for the normal hearing processes. In this study we examined the localization of PAC1 receptor and Ca(2+) buffering proteins (parvalbumin, calretinin, calbindin) in the inner ear of 5-day-old PACAP-deficient mice compared with wild-type mice in order to get a closer insight into the effect of endogenous PACAP in the cochlear function. We did not find differences in the distribution pattern of PAC1 receptors between the two groups, but wild-type animals showed significantly higher PAC1 receptor expression. In contrast, inner and outer hair cells of PACAP-deficient mice showed more pronounced parvalbumin, calbindin, and calretinin immunopositivity compared with wild-type mice. Elevated endolymphatic Ca(2+) is deleterious for cochlear function, while the high concentration of Ca(2+) buffers in hair cells may offer protection. The increased immunoreactivity of Ca(2+) binding proteins in the absence of PACAP provide further evidence the important role of PACAP in the hearing processes.

Role of PACAP in Neural Stem/Progenitor Cell and Astrocyte: from Neural Development to Neural Repair

After central nervous system (CNS) injury, reactive astrocytes display opposing functions, inducing neural repair and axonal regeneration via the release of growth factors, or forming a glial scar which acts as a barrier to axonal regeneration. Endogenous neural stem/progenitor cells have also recently been identified at the site of CNS injury, where they have been shown to differentiate into mature neurons in an animal model of ischemia. However, the pathophysiological mechanisms underpinning the contribution of reactive astrocytes and neural stem/progenitor cells to neural repair are still to be fully elucidated. Pituitary adenylate cyclase activating polypeptide (PACAP) is widely expressed in the CNS, where it has been shown to exert numerous biological effects. This review will summarize the current state of knowledge regarding the expression of PACAP and its receptors during neural development, as well as the involvement of PACAP in astrocytes and neural stem/progenitor cell biology. In addition, we will also discuss emerging evidence that implicates PACAP in neurogenesis and neural repair in response to brain pathophysiology.

Possible involvement of PACAP and PACAP type 1 receptor in GnRH-induced FSH β-subunit gene expression

Pituitary adenylate cyclase-activating polypeptide (PACAP) and its receptor, PACAP type 1 receptor (PAC1-R) play an important role in the induction of pituitary gonadotropins. In this present study, we examined whether the PAC1-R was involved in the action of gonadotropin-releasing hormone (GnRH) on gonadotropin FSHβ subunit expression. In a static culture, GnRH stimulation significantly increased PAC1-R expression as well as PACAP gene expression in the gonadotroph cell line, LβT2. Stimulation with low frequency GnRH pulses, which preferentially increase FSHβ, increased the expression of both the PAC1-R and the PACAP genes to a greater extent than did high frequency pulses.In the determination of transcriptional activity, the GnRH antagonist, cetrotide inhibited GnRH-induced FSHβ promoter activity completely, but PACAP6-38, a PACAP antagonist, had no effect on GnRH-induced FSHβ promoter activity. As expected, PACAP-induced FSHβ promoter activity was significantly prevented by PACAP6-38, but was not affected by cetrotide. PACAP6-38, however, significantly prevented GnRH-increased FSHβ mRNA expression. These observations suggest that GnRH-induced FSHβ gene expression is stimulated partially through PAC1-R by gonadotrophs producing PACAP or PAC1-R.

Comparative immunhistological examination of inner ear in wild type and pituitary adenylate cyclase activating polypeptide (PACAP) deficient mice

Event Abstract Back to Event Comparative immunhistological examination of inner ear in wild type and pituitary adenylate cyclase activating polypeptide (PACAP) deficient mice A. Tamás1, B. Fülöp1, D. Reglődi1, P. Kiss1, R. Brubel1, G. Horváth1*, A. Lubics1, R. Gábriel2 and K. Szabadfi2 1 University of Pécs, Department of Anatomy, Medical School, Institute of Biology, Hungary 2 University of Pécs, Department of Experimental Zoology and Neurobiology, Institute of Biology, Hungary PACAP is a multifunctional neuropeptide with well known neuroprotective and neurotrophic effects. The involvement of PACAP in sensory processing has also been documented, but little is known about its effects in the auditory system. PACAP and its specific receptor (PAC1), are present in the cochlea and in brain structures involved in auditory pathways. Recently, we have shown that PACAP protects cochlear cells against oxidative stress-induced apoptosis. The endolymphatic Ca2+ cc. controlled by Ca2+ buffers of the hair cells is essential for the normal hearing processes. In this study we examined the localization of PAC1-receptor and Ca2+ buffering proteins (parvalbumin, calretinin) in the inner ear of 5-day-old PACAP deficient mice compared with wild type mice to investigate the effect of endogenous PACAP in the cochlear function. We did not find difference in the distribution of PAC1 receptors between groups. In contrast, inner and outer hair cells of PACAP KO mice showed more pronounced parvalbumin and calretinin immunpositivity compared with wild type mice. Elevated endolymphatic Ca2+ is deleterious for the cochlear function, against which the high cc. of Ca2+ buffers in hair cells may protect. Meanwhile, the increased immunreactivity of Ca2+ binding proteins in the absence of PACAP provide further evidence the important role of PACAP in the hearing processes. Acknowledgements Support: OTKAK72592, F67830, CNK78480, ETT278-04/2009, Bolyai Scholarship,Richter Foundation Keywords: Molecular and cellular neurobiology, Neuroscience Conference: 13th Conference of the Hungarian Neuroscience Society (MITT), Budapest, Hungary, 20 Jan - 22 Jan, 2011. Presentation Type: Abstract Topic: Molecular and cellular neurobiology Citation: Tamás A, Fülöp B, Reglődi D, Kiss P, Brubel R, Horváth G, Lubics A, Gábriel R and Szabadfi K (2011). Comparative immunhistological examination of inner ear in wild type and pituitary adenylate cyclase activating polypeptide (PACAP) deficient mice. Front. Neurosci. Conference Abstract: 13th Conference of the Hungarian Neuroscience Society (MITT). doi: 10.3389/conf.fnins.2011.84.00053 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 03 Mar 2011; Published Online: 23 Mar 2011. * Correspondence: Dr. G. Horváth, University of Pécs, Department of Anatomy, Medical School, Institute of Biology, Pécs, Hungary, horvath@phys.szote.u-szeged.hu Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers A. Tamás B. Fülöp D. Reglődi P. Kiss R. Brubel G. Horváth A. Lubics R. Gábriel K. Szabadfi Google A. Tamás B. Fülöp D. Reglődi P. Kiss R. Brubel G. Horváth A. Lubics R. Gábriel K. Szabadfi Google Scholar A. Tamás B. Fülöp D. Reglődi P. Kiss R. Brubel G. Horváth A. Lubics R. Gábriel K. Szabadfi PubMed A. Tamás B. Fülöp D. Reglődi P. Kiss R. Brubel G. Horváth A. Lubics R. Gábriel K. Szabadfi Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.

PACAP ameliorates oxidative stress in the chicken inner ear: An in vitro study

Pituitary adenylate cyclase activating polypeptide (PACAP) is a pleiotropic and multifunctional neuropeptide. Numerous studies prove that PACAP has neuroprotective effects in diverse neuronal systems in vitro and in vivo. The involvement of PACAP in visual and olfactory sensory processing has also been documented, but little is known about its effects in the auditory system. The presence of PACAP and its receptor, the specific PAC1 receptor, has been shown in the cochlea and in brain structures involved in auditory pathways. The aim of the present study was to investigate whether PACAP is protective in cochlear oxidative stress-induced cell death, which is known to play a role in several ototoxic insults. Chicken cochlear cells were exposed to 1 mM H 2O 2, which resulted in a marked reduction of cell viability and a parallel increase of apoptotic and necrotic cells assessed by MTT test, annexin V/propidium iodide flow cytometry and JC-1 apoptosis assay. Co-incubation with 100 nM PACAP increased cell viability and reduced the percentage of apoptotic cells. Furthermore, oxidative stress increased the activation of caspase-3, while simultaneous PACAP treatment reduced it. In summary, our present results demonstrate that PACAP effectively protects cochlear cells against oxidative stress-induced apoptotic cell death.

Progesterone increases mRNA levels of pituitary adenylate cyclase-activating polypeptide (PACAP) and type I PACAP receptor (PAC 1) in the rat hypothalamus

Pituitary adenylate cyclase-activating polypeptide (PACAP) regulates pituitary hormone biosynthesis and secretion through its cognate receptors. PACAP also plays an important role in the regulation of ovarian steroid biosynthesis. If so, there might be a feedback regulation of hypothalamic PACAP synthesis by the pituitary and by ovarian steroids. In the present study, we used RNase protection assays to determine changes in mRNA levels of PACAP and type I PACAP receptor (PAC 1) under the conditions of ovariectomy and replacement with ovarian steroids. Progesterone (P) alone or in combination with estradiol (E) induced significant increases in PACAP mRNA level in the medial basal hypothalamus (MBH) and PAC 1 mRNA levels in MBH and the preoptic area (POA). This finding suggests that feedback regulation takes place between the ovary and hypothalamic PACAP neurons. P is known to be a major regulatory feedback factor for hypothalamic luteinizing hormone-releasing hormone (LHRH) neurons, but P receptor is not present in these neurons. Therefore, we examined a possible involvement of PACAP in the feedback regulatory pathway of P to LHRH neurons. After an antisense PAC 1 oligodeoxynucleotide (ODN) was i.c.v.-injected into the third ventricle of E and P-treated rats, LHRH mRNA levels were determined. The ODN markedly decreased the P-induced increase in the LHRH mRNA level. Taken together, the present data suggest that PACAP may play a role as a mediator in the regulation of LHRH synthetic machinery by stimulatory feedback of P.

Localization, characterization and activity of pituitary adenylate cyclase-activating polypeptide in the frog adrenal gland.

Pituitary adenylate cyclase-activating polypeptide (PACAP) has recently been isolated from the frog brain and the sequence of the peptide appears to be strikingly similar to that of mammalian PACAP. In the present study, we have investigated the localization of PACAP in the frog interrenal (adrenal) gland by immunocytochemistry using antisera directed against PACAP 38 or PACAP 27. Two types of PACAP-immunoreactive fibres were observed: thick varicose fibres coursing between adrenal cells and thin processes located in the walls of blood vessels irrigating the gland. Bilateral transection of the splanchnic nerves did not affect the intensity and distribution of PACAP immunoreactivity. The mean +/- S.E.M. concentration of PACAP, measured by radioimmunoassay in crude adrenal extracts, was 0.65 +/- 0.16 nmol/g wet tissue. Two molecular forms of PACAP in the adrenal gland were characterized by reversed phase high-performance liquid chromatography combined with radioimmunoassay quantification. The elution profiles revealed the existence of two peaks exhibiting the same retention times as synthetic frog PACAP 38 (fPACAP 38) and PACAP 27, the predominant form being PACAP 38. The possible involvement of PACAP in the regulation of adrenal steroidogenesis was investigated in vitro using a perifusion system for frog adrenal slices. Graded doses of fPACAP 38 (0.1-10 mumol/l) increased the secretion of both corticosterone and aldosterone in a dose-dependent manner. Administration of repeated pulses of fPACAP 38 (1 mumol/l), at 120-min intervals, led to a reproducible stimulation of corticosteroid secretion without any tachyphylaxis. Prolonged infusion (2 h) of the peptide induced a rapid increase in corticosterone and aldosterone output, followed by a gradual decline in the secretion rate, suggesting the occurrence of a desensitization phenomenon. Synthetic porcine vasoactive intestinal peptide, which is structurally related to PACAP, was about ten times less potent than fPACAP 38 in stimulating steroidogenesis while the [Des-His1]-fPACAP 38 analogue was 100 times less effective. These results demonstrate that a peptide closely related to fPACAP 38 is present in fibres innervating the frog adrenal gland and could participate in the regulation of corticosteroid secretion, particularly during neurogenic stress.