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

ABSTRACT 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 μmol/l) increased the secretion of both corticosterone and aldosterone in a dose-dependent manner. Administration of repeated pulses of fPACAP 38 (1 μmol/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. Journal of Endocrinology (1993) 139, 183–194

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PITUITARY ADENYLATE CYCLASE-ACTIVATING PEPTIDE-38 (PACAP)-38 IS RELEASED INTO HYPOPHYSIAL PORTAL BLOOD IN THE NORMAL MALE AND FEMALE RAT

Journal of Endocrinology ◽

10.1677/joe.0.142r001 ◽

1994 ◽

Vol 142 (1) ◽

pp. R1-R4 ◽

Cited By ~ 72

Author(s):

R C Dow ◽

J Bennie ◽

G Fink

Keyword(s):

Adenylate Cyclase ◽

High Performance ◽

Reversed Phase ◽

Portal Blood ◽

Molecular Forms ◽

Normal Male ◽

Female Rat ◽

Pituitary Adenylate Cyclase ◽

Amidated Peptide ◽

Hypophysial Portal

ABSTRACT Pituitary adenylate cyclase activating polypeptide (PACAP) was first isolated from ovine hypothalamus as two molecular forms, the basic 38 residue amidated peptide PACAP-38 and the N-terminal 27 amino acid sequence PACAP-27. A dense plexus of PACAP immunoreactive fibres is present in the internal and external layers of the median eminence and in other parts of the hypothalamus with PACAP cell bodies in the paraventricular and supraoptic nuclei. The present study shows, for the first time, that, as assessed by radioimmunoassay of extracted plasma, the amount of PACAP-38 in hypophysial portal is significantly greater than in peripheral blood, and that as assessed by reversed phase high performance liquid chromatography, PACAP 1-38 is the major form in portal blood. This evidence is crucial for the fact that PACAP-38 may be a hypothalamic-pituitary regulatory factor.

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

Journal of Endocrinology ◽

10.1677/joe-07-0127 ◽

2007 ◽

Vol 196 (2) ◽

pp. 291-303 ◽

Cited By ~ 10

Author(s):

Salvatore Valiante ◽

Marina Prisco ◽

Rosaria Sciarrillo ◽

Maria De Falco ◽

Anna Capaldo ◽

...

Keyword(s):

Adrenal Gland ◽

Adenylate Cyclase ◽

Vasoactive Intestinal Polypeptide ◽

Chromaffin Cells ◽

Adrenal Glands ◽

Podarcis Sicula ◽

Adrenal Cell ◽

Adrenal Cells ◽

Pituitary Adenylate Cyclase

Vasoactive intestinal polypeptide (VIP) and pituitary adenylate cyclase-activating polypeptide (PACAP) are regulatory neuropeptides of the hypothalamus–hypophyseal–adrenal axis, acting via the common receptors VPAC1 and VPAC2 and the selective PACAP receptor PAC1. In the adrenal glands of the Italian wall lizard, Podarcis sicula, the presence of VIP in chromaffin cells, and the VIP-stimulated release of catecholamine and aldosterone in vivo, was previously shown. To examine the localization of both peptides and receptors and their mRNAs in the adrenal gland of P. sicula, immunohistochemistry and in situ hybridization were performed: PACAP and its mRNA were detected in chromaffin cells, VPAC1 was found associated with steroidogenic tissue, VPAC2 and PAC1 with chromaffin tissue. Using ‘far western blot’ technique, we showed the presence of specific binding sites for VIP/PACAP in the adrenal glands of the lizard. The effects of both VIP and PACAP on the adrenal cells of the lizard were examined in vitro in adrenal cell co-cultures: both VIP and PACAP enhanced catecholamine, corticosterone and aldosterone release from adrenal cell co-culture in a time- and dose-dependent manner. The catecholamine release was inhibited by PAC1 antagonist and in VPAC2 immunoneutralized adrenal cells. The effects of VIP and PACAP on aldosterone secretion were counteracted by VPAC1 antagonist administration in vitro. Corticosterone secretion elicited by VIP was not blocked by VPAC1 antagonist, while the PACAP-induced release of corticosterone was blocked by the antagonist. Overall, our investigations indicate that these neuropeptides of the secretin superfamily can act not only as neurotransmitters but also as autocrine and paracrine regulators on chromaffin and cortical cells, being important mediators of the non-cholinergic system in the lizard adrenal gland.

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Involvement of the cytoskeleton in the steroidogenic response of frog adrenal glands to angiotensin II, acetylcholine and serotonin

Journal of Endocrinology ◽

10.1677/joe.0.1180365 ◽

1988 ◽

Vol 118 (3) ◽

pp. 365-NP ◽

Cited By ~ 31

Author(s):

M. Feuilloley ◽

P. Netchitaïlo ◽

C. Delarue ◽

F. Leboulenger ◽

M. Benyamina ◽

...

Keyword(s):

Angiotensin Ii ◽

High Performance ◽

Cytochalasin B ◽

Adrenal Steroidogenesis ◽

Microtubular Network ◽

Spontaneous Secretion ◽

Corticosteroid Secretion ◽

Stimulation Of

ABSTRACT In order to determine the role of the cytoskeleton in adrenal steroidogenesis, we have studied the effect of cytochalasin B (a microfilament-disrupting agent) and vinblastine (an antimicrotubular drug) on corticosteroid secretion by frog interrenal tissue in vitro. Perifusion of interrenal fragments with cytochalasin B (50 μmol/l) induced a marked inhibition of basal corticosteroid output. In addition, stimulation of corticosteroidogenesis by all corticotrophic factors was also inhibited by cytochalasin B. Using an immunohistochemical technique and specific anti-tubulin antiserum, we verified that vinblastine (10 μmol/l) was responsible for the disappearance of the microtubular network in adrenocortical cells. Administration of vinblastine (10 μmol/l) did not affect the spontaneous secretion of corticosterone and aldosterone and had no effect on the steroidogenic response of interrenal glands to angiotensin II and acetylcholine. In contrast, vinblastine was responsible for a marked decrease in serotonin-induced stimulation of corticosteroid production. On the other hand, data from high-performance liquid chromatography showed that infusion of cytochalasin B or vinblastine was not associated with the production of any new steroid which could interfere in the radioimmunoassays. Taken together, these data suggest that microfilaments are involved in a late and common step of corticosteroidogenesis while microtubules are only required for the coupling of the secretory response to certain corticotrophic factors such as ACTH and serotonin. J. Endocr. (1988) 118, 365–374

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Accumulation of rat pineal serotonin N-acetyltransferase mRNA induced by pituitary adenylate cyclase activating polypeptide and vasoactive intestinal peptide in vitro

Journal of Molecular Endocrinology ◽

10.1677/jme.0.0280019 ◽

2002 ◽

Vol 28 (1) ◽

pp. 19-31 ◽

Cited By ~ 8

Author(s):

Z Rekasi ◽

T Czompoly

Keyword(s):

Gene Expression ◽

Adenylate Cyclase ◽

Vasoactive Intestinal Peptide ◽

Mrna Level ◽

Mrna Levels ◽

Dependent Manner ◽

Mrna Accumulation ◽

Reverse Transcription Pcr ◽

Pituitary Adenylate Cyclase

In mammals, pineal melatonin secretion is under the control of adrenergic and peptidergic inputs regulating serotonin N-acetyltransferase (arylalkylamine N-acetyltransferase; AA-NAT) activity. In this study, the accumulation of AA-NAT mRNA induced by norepinephrine (NE) and peptides of the secretin superfamily (pituitary adenylate cyclase activating polypeptide (PACAP), vasoactive intestinal peptide (VIP), growth hormone releasing factor (GRF), secretin) was investigated by a new quantitative reverse transcription-PCR (RT-PCR) assay. We demonstrated that PACAP was the most potent peptide to increase the expression of AA-NAT mRNA and to induce cAMP production in rat pinealocytes. VIP was also able to elevate the AA-NAT mRNA level and cAMP efflux in a dose-dependent manner; however, it was six- and threefold, respectively, less potent than PACAP. The maximal values of AA-NAT mRNA level after PACAP and VIP exposures were similar (523.1 +/- 52.5 amol to 640.7 +/- 68.8 amol vs 461.5 +/- 54.3 amol to 579.2 +/- 72.4 amol). These saturable peak values were approximately five- to eightfold less than that after NE (3.0 +/- 0.3 fmol to 3.6 +/- 0.4fmol). GRF and secretin were less potent than VIP in inducing AA-NAT gene expression and cAMP efflux. These data suggest that the peptides act mostly on VIP(1)/PACAP (VPAC(1)) receptors of pinealocytes with different affinity. The peak cAMP efflux always preceded the elevation of AA-NAT gene expression during the 3-h infusion of VIP or NE. The cAMP efflux had declined by the time of onset of maximal AA-NAT gene expression, but remained significantly higher than its basal values. Our data indicate that even a submaximal level of cAMP is sufficient for maintaining the maximal AA-NAT mRNA accumulation. These findings show that, in addition to NE, PACAP and VIP may have an important role in the regulation of AA-NAT mRNA levels in rat pinealocytes.

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In Vitro Evaluation of Various Buccal Permeation Enhancing Systems for PACAP (Pituitary Adenylate Cyclase-Activating Polypeptide)

Pharmaceutical Research ◽

10.1007/s11095-005-7894-4 ◽

2005 ◽

Vol 22 (12) ◽

pp. 2045-2050 ◽

Cited By ~ 27

Author(s):

Nina Langoth ◽

Andreas Bernkop-Schnürch ◽

Peter Kurka

Keyword(s):

Adenylate Cyclase ◽

In Vitro Evaluation ◽

Pituitary Adenylate Cyclase

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Pituitary adenylate cyclase-activating polypeptide and PACAP receptor expression and function in the rat adrenal gland

International Journal of Molecular Medicine ◽

10.3892/ijmm.9.3.233 ◽

2002 ◽

Cited By ~ 2

Author(s):

Giuseppina Mazzocchi ◽

Ludwik Malendowicz ◽

Giuliano Neri ◽

Paola Andreis ◽

Agnieszka Ziolkowska ◽

...

Keyword(s):

Adrenal Gland ◽

Adenylate Cyclase ◽

Receptor Expression ◽

Pituitary Adenylate Cyclase ◽

And Function

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The use of pituitary adenylate cyclase-activating polypeptide in the pre-maturation system improves in vitro developmental competence from small follicles of porcine oocytes

Asian-Australasian Journal of Animal Sciences ◽

10.5713/ajas.19.0162 ◽

2019 ◽

Vol 32 (12) ◽

pp. 1844-1853

Author(s):

Kyu-Mi Park ◽

Kyu-Jun Kim ◽

Minghui Jin ◽

Yongquan Han ◽

Kyoung-Ha So ◽

...

Keyword(s):

Adenylate Cyclase ◽

Developmental Competence ◽

Pituitary Adenylate Cyclase

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Formulation, characterization, evaluation and in vitro study of transfersomal gel medroxyprogesterone acetate for transdermal drug delivery

International Journal of Research in Pharmaceutical Sciences ◽

10.26452/ijrps.v11i4.3159 ◽

2020 ◽

Vol 11 (4) ◽

pp. 5373-5381

Author(s):

Iskandarsyah ◽

Camelia Dwi Putri Masrijal ◽

Harmita

Keyword(s):

Drug Delivery ◽

Medroxyprogesterone Acetate ◽

Transdermal Drug Delivery ◽

High Performance ◽

Hormonal Contraception ◽

In Vitro Study ◽

Entrapment Efficiency ◽

Tween 80 ◽

Reversed Phase

A hormonal contraception progestin such as medroxyprogesterone acetate (MPA) is used to helps regulate ovulation thus as a part of contraception hormone therapy as a method of birth control. This study aimed to formulate, characterized, evaluated transfersomal gel containing medroxyprogesterone acetate and to increased subcutaneous penetration of medroxyprogesterone acetate. In this research, three transfersomes formulas were prepared and optimized, e.g. F1, F2 and F3 with phosphatidylcholine: tween 80 concentration were 90:10; 85:15; and 75:25, respectively. F2 was the best formula with the highest entrapment efficiency 81.20±0.42 %, Average 81.35 ±0.78 nm, morphology of vesicles were spheres, indeks polidispersity 0.198±0.012 and zeta potential was -34.83±0.64 mV. The transpersonal gel (FGT) containing F2, and non-transpersonal gel containing MPA in methanol(FG) were prepared. In vitro penetration test were conducted to both of them using Franz Diffusion cells. Analysis of medroxyprogesterone acetate used a high performance liquid chromatographic (HPLC) method with an ultraviolet detector on reversed-phase C18, 5µm; 150 x 4.6 mmcolumn; using acetonitrile-0.1% formic acid (60:40/v:v) and was detected at a wavelength of 240 nm with flow rate at 1.0 mL/min. Gel stability evaluation results showed that FGT was better than FG on pH stability, viscosity and rheological properties. Based on in vitro penetration study, cumulative subcutaneous penetration of medroxyprogesterone acetate from FGT was 2356.45 ± 197.73 ng.cm-2 and from FG 359.15 ± 13.60 ng.cm-2, respectively. Flux value for FGT and FG were 112.77 ± 6,47 ng.cm-2.hr-1and 17.99 ± 4.81 ng.cm-2.hr-1, respectively. It could be concluded that transfersomal gel medroxyprogesterone acetate for transdermal drug delivery increased cumulative transdermal penetration of medroxyprogesterone acetate by six times more than non-transfersomal gel dosage form.

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