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At least 19 recordsLinked to original sources

Regulatory peptides--the distribution of two newly discovered peptides: PHI and NPY.

PHI (peptide with histidine and isoleucine) and NPY (neuropeptide with tyrosine) are recently discovered regulatory peptides. There are close sequence similarities between PHI and both growth hormone releasing factor (GRF) and vasoactive intestinal polypeptide (VIP) and between NPY and pancreatic polypeptide. Using immunocytochemistry and radioimmunoassay we have revealed the broad distributions of PHI and NPY in neurons of the central nervous system and the majority of peripheral tissues. Tissues which are particularly well provided with these peptides include gut, pancreas, respiratory tract, skin and the genitourinary and cardiovascular systems. In most peripheral tissues, PHI-containing ganglion cells occur locally. NPY-containing fibres originate in part from cell bodies outside the tissues, in the sympathetic nervous system. Comparative studies indicate that PHI and VIP are co-stored in the same neurone and are identically distributed, thus suggesting the existence of a common precursor and subsequent gene duplication. The possible co-existence of catecholamines and NPY, suggested by the consistent finding of very similar distributions of the two substances, was investigated using antibodies to converting enzymes involved in catecholamine synthesis (tyrosine-hydroxylase and dopamine-beta-hydroxylase). The two enzymes and NPY were found together in at least part of the same neuronal system.

Animals↗

Isolation of a brain peptide identical to the intestinal PHI (peptide HI).

The isolation of a brain peptide identical to the intestinal peptide PHI (peptide HI) is described. The peptide was isolated from porcine brain extract using a chemical assay method based on its C-terminal isoleucine amide structure. The complete amino acid sequence of the peptide was found to be: His-Ala-Asp-Gly-Val-Phe-Thr-Ser-Asp-Phe-Ser-Arg-Leu-Leu-Gly-Gln-Leu-Ser-Ala- Lys-Lys-Tyr-Leu-Glu-Ser-Leu-Ile-NH2. This sequence is identical to the intestinal peptide thus demonstrating PHI to be a brain-gut peptide. The role of PHI in the central nervous system as a neurotransmitter or neuromodulator is discussed.

Amino Acid Sequence↗

Prolactin-releasing activity of porcine intestinal peptide (PHI-27).

Porcine intestinal peptide (PHI), a twenty-seven amino acid peptide isolated from porcine gut extracts, is a close structural homolog of the secretin family hormones. The structural and biological similarities of PHI to vasoactive intestinal peptide (VIP) together with its presence in the rat hypothalamus suggested a possible role for the peptide in the control of prolactin (PRL) secretion. PHI induced significant, dose-related stimulations of PRL release from cultured, dispersed rat pituitary cells in vitro. The minimum effective dose is 10(-7) molar, compared to 10(-9) molar for VIP. No interactive effect with thyrotropin-releasing hormone was observed; however, PHI partially overcame the dopamine inhibition of PRL release.

Animals↗

Effect of the peptide PHI-27 on prolactin release in vitro.

The present study demonstrates that PHI, a peptide belonging to the glucagon-secretin group and thus structurally similar to VIP, can release prolactin from dispersed rat anterior pituitary cells and also causes release of prolactin from hemipituitaries. PHI-like immunoreactivity has previously been demonstrated in a hypothalamic system with nerve endings in the median eminence, and, taken together, these findings suggest that PHI may represent a physiologic prolactin-releasing factor.

Animals↗

Actions of a newly isolated intestinal peptide PHI on pancreatic acini.

In dispersed acini from guinea pig pancreas, PHI, a peptide recently isolated from porcine intestine and found to contain 27 amino acids, inhibited binding of 125I-vasoactive intestinal peptide (125I-VIP), increased cellular cAMP, and stimulated amylase secretion. The increase in amylase secretion caused by a maximally effective concentration of PHI in combination with 8-bromo-cAMP, VIP, or secretin was the same as that caused by PHI alone. In contrast, the increase in amylase secretion caused by PHI plus bombesin, carbachol, or the C-terminal octapeptide of cholecystokinin was significantly greater than the sum of the increase caused by each secretagogue acting alone. From the abilities of PHI to inhibit binding of 125I-VIP, to increase cellular cAMP, and to increase amylase secretion, the apparent affinity of PHI for the VIP-preferring receptors on pancreatic acinar cells is approximately 25 times less than that of VIP but 10 times greater than that of secretin. From the ability of PHI to increase cellular cAMP, the apparent affinity of PHI for the secretin-preferring receptors on pancreatic acinar cells is approximately 300 times less than that of secretin but equal to that of VIP.

Amylases↗

Isolation and primary structure of human PHI (peptide HI).

The isolation of the human form of PHI (peptide HI) is described. The peptide was purified from human colonic extracts by using a chemical method for the detection of its C-terminal amidated structure. Human PHI consists of 27 amino acid residues and the complete amino acid sequence is: His-Ala-Asp-Gly-Val-Phe-Thr-Ser-Asp-Phe-Ser-Lys-Leu-Leu-Gly-Gln-Leu-Ser- Ala-Lys-Lys-Tyr-Leu-Glu-Ser-Leu-Met-NH2. The differences between the structures of porcine and human PHI are at position 12 (Arg/Lys replacement) and at position 27 (Ile/Met).

Amino Acid Sequence↗

Determination of peptide phi angles in solids by relayed anisotropy correlation NMR.

A solid state NMR method is presented for determination of a backbone dihedral angle phi in peptides, being based on the previously reported method, relayed anisotropy correlation (RACO) NMR [Y. Ishii et al., Chem. Phys. Lett. 256 (1996) 133]. In the present method, the 15N-1H and the 13C-1H dipolar tensors in the 1H-15N-13C-1H system are two-dimensionally (2D) correlated via polarization transfer from 15N to 13C under magic angle spinning (MAS). This method was applied to N-acetyl[1,2-13C,15N]D,L-valine, and the H-C-N-H dihedral angle was determined to be 154.0 +/- 1.4 degrees or 206.0 +/- 1.4 degrees, the former agreeing with the X-ray value of 154 +/- 5 degrees.

Anisotropy↗

Stimulatory effect of the intestinal peptide PHI on glycogenolysis and gluconeogenesis in isolated rat hepatocytes.

The newly isolated peptide PHI provoked a dose-dependent stimulation of glycogenolysis and gluconeogenesis in isolated rat hepatocytes; at 1 microM-PHI, both processes were increased 1.6-fold as compared with basal values. These PHI-mediated effects were accompanied by the activation of glycogen phosphorylase and the inactivation of pyruvate kinase. PHI (1 microM) also caused a 2-fold increase in hepatocyte cyclic AMP.

Animals↗

The distribution and chromatographic characterization of PHI (peptide histidine isoleucine amide)-27-like peptides in rat and porcine brain.

This study was initiated to characterize PHI (peptide histidine isoleucine amide)-27-like peptides (PLPs) in rat and porcine brain in comparison with other members of the vasoactive intestinal polypeptide (VIP) family and to investigate their distribution by radioimmunoassay. The peptidic nature of the rat brain PLP was indicated by its trypsin sensitivity. On Sephadex chromatography rat brain PLP has the same molecular weight as synthetic (porcine intestinal) PHI-27. High pressure liquid chromatographic separations revealed that PLP in rat and porcine brain extracts elutes as a single peak distinct from VIP or secretin. Porcine brain PLP elutes in the same position as synthetic PHI-27, whereas rat brain PLP immunoreactivity consistently separates from synthetic PHI-27. This suggests that porcine brain PLP is identical to synthetic PHI-27, in agreement with the reported sequence of Tatemoto et al. (Tatemoto, K., M. Carlquist, T. McDonald, and V. Mutt (1983) FEBS Lett. 153: 248-252), whereas PLP may have a different amino acid sequence (or may be post-translationally modified). Using specific PHI and VIP radioimmunoassays, the distribution of PLP was found to parallel that of VIP in rat and porcine brain, being highest in cerebral cortex, amygdala, and hippocampus. PLP, like VIP, is abundant in rat retina and can be included in the growing list of retinal peptides. This highly correlated distribution of VIP and PLP may be explained by the recent discovery that they are derived from the same precursor (Itoh, N., K. Obata, N. Yanaihara, and H. Okamoto (1983) Nature 304: 547-549).(ABSTRACT TRUNCATED AT 250 WORDS)

Amygdala↗

Absence of effect of the peptide PHI-27 on LH release in vitro.

The effects of PHI-27, a peptide of the glucagon-secretion family, on luteinizing hormone (LH) release and on LH-releasing hormone (LH-RH)- or estradiol-induced LH release were examined in a sequential double chamber perifusion system by perifusing the pituitary alone or in sequence with the mediobasal hypothalamus (MBH) from normal female rats in diestrus. PHI at 10(-7) M had no significant effect on LH release from the pituitary in series with the MBH. Moreover, on perifusion of the pituitary alone with medium containing 10(-7) M PHI, LH release induced by 20 ng/ml LH-RH from the pituitary was not significantly different from that without PHI. Furthermore, PHI had no effect on estradiol-induced LH release from the pituitary in sequence with the MBH. These data indicate that PHI has no effect on LH release in vitro.

Animals↗

Stimulation of growth hormone release by vasoactive intestinal peptide and peptide PHI in rat anterior pituitary reaggregates. Permissive action of a glucocorticoid and inhibition by thyrotropin-releasing hormone.

In superfused rat anterior pituitary cell reaggregates, cultured for 5 days in serum-free defined medium, vasoactive intestinal peptide (VIP) concentration-dependently stimulated prolactin (Prl) release but had only a marginal effect on growth hormone (GH) release. When reaggregates were cultured in the presence of 80 nM dexamethasone (Dex) VIP strongly stimulated GH release from a concentration as low as 0.1 nM. VIP did not stimulate LH release. Peptide PHI also stimulated GH release but thyrotropin-releasing hormone (TRH) or angiotensin II did not. In fact, TRH slightly but transiently inhibited basal GH release and strongly inhibited VIP-stimulated GH release. GH-releasing factor (GRF) stimulated GH more potently and with higher intrinsic activity than VIP but GRF did not increase Prl release. The present data indicate that under defined hormonal conditions VIP and PHI are capable of stimulating GH release and that TRH can antagonize this effect by a direct action on the pituitary.

Animals↗

The intestinal peptide PHI-27 potentiates the action of corticotropin-releasing factor on ACTH release from rat pituitary fragments in vitro.

Effects of rat corticotropin releasing factor (r-CRF) and of natural and synthetic porcine PHI-27 on ACTH release were studied by using freshly prepared anterior pituitary fragments of rats that were superfused with Krebs Ringer bicarbonate buffer (KRB). ACTH was measured in effluent medium samples by RIA. Superfusion with r-CRF-containing KRB caused a concentration-dependent (range 0.2-10 mM) increase of ACTH release. PHI alone did not alter the spontaneous release of ACTH at concentrations of up to 700 nM, but 2000 nM of natural or synthetic porcine PHI-27 enhanced the release rate of ACTH to 180 and 150% of basal ACTH release respectively. Addition of a subeffective concentration of PHI (300nM) to r-CRF containing KRB strongly potentiated its ACTH-releasing effect. Since PHI immunoreactive material is present in nerve fibres located in the external zone of the median eminence, we suggest that PHI or related products may play a physiological role in the control of ACTH secretion.

Adrenocorticotropic Hormone↗

Infusion of a novel peptide, PHI, in man. Pharmacokinetics and effect on gastric secretion.

PHI, infused in man, achieved plateau plasma levels of 297 pmoles/1. The plasma half life was 3.1 min, metabolic clearance rate was 16.4 ml/kg/min and estimated volume of distribution was 73.2 ml/kg. No subjective side effects were noted during the infusion and there was no significant alteration in submaximal pentagastrin stimulated gastric acid or pepsin secretion.

Adult↗

Peptides PHI and VIP: comparison between vascular and nonvascular smooth muscle effect in rabbit uterus.

The distribution and effects of the two neuropeptides, vasoactive intestinal polypeptide (VIP) and peptide histidine isoleucine amide (PHI), on vascular and nonvascular smooth muscle in the urogenital tract of nonpregnant rabbit female, were investigated. Immunoreactive VIP and PHI were present in all regions except the ovary with the highest concentration in the uterine cervix. By using in vitro tension recordings of myometrial specimens, it was demonstrated that both peptides displayed a dose-dependent inhibition of the mechanical activity. The dose-response curves of VIP and PHI were superimposable with and ID50 of 3 X 10(-8) mol/l, and their combined effect was additive. In addition, the influence of the two peptides on myometrial blood flow (MBF) was investigated by the xenon-133 washout technique. Both peptides were found to increase MBF with the same potency and efficacy. Their combined effect was additive. In conclusion VIP and PHI are present in the rabbit urogenital tract, and the two peptides are equipotent inhibitors of mechanical nonvascular and vascular smooth muscle activity in the uterus.

Animals↗

Synthesis of the porcine intestinal peptide PHI and its 24-glutamine analogue.

PHI--a new candidate hormone from porcine intestinal tract-- corresponds to a linear heptacosapeptide amide of remarkable sequence homology to the known members of the glucagon family, particularly to the vasoactive intestinal peptide (VIP) and secretin. The position 24 usually occupied by an aminodicarboxylic acid omega-amide, in the present case, however, carries a glutamic acid, thus opening the question of whether this structural feature is related to desamidation in one of the isolation and characterization steps or of whether it is significant for this peptide factor. Consequently the heptacosapeptide amides corresponding to the proposed primary structure and to its 24-glutamine analogue have been synthesized. Comparative chromatographic and biological studies on the natural and the two synthetic products have confirmed the correctness of the primary structure proposed for the isolated PHI. Since [24-glutamic acid] and [24-glutamine]PHI exhibit no significant differences in their biological potencies, the main question is still open of whether the position 24 in native PHI is occupied by the aminodicarboxylic acid omega-amide (glutamine) or by N-substituted derivatives (N-glycosyl).

Amino Acid Sequence↗

Concentrations and distribution of vasoactive intestinal peptide (VIP), peptide histidine isoleucine (PHI) and peptide histidine valine (PHV) in the cerebral cortex and the suprachiasmatic nucleus of the mouse.

Prepro-vasoactive intestinal peptide (prepro-VIP) is processed to at least three biologically active peptides: VIP, peptide histidine isoleucine (PHI) and an extended PHI, peptide histidine valine (PHV). The aim of the present investigation was by chromatography combined with RIA and immunocytochemistry to determine which of these peptides were present in the cerebral cortex and the hypothalamic suprachiasmatic nucleus (SCN) of the mouse. These regions were chosen since they are known to contain a high concentration of VIP but the relative concentration of PHI and PHV is not known. Tissue was extracted and subjected to gel chromatography and high-pressure liquid chromatography (HPLC). VIP and PHI immunoreactivities co-eluted with synthetic rat VIP and PHI. A minor peak of PHI and prepro-VIP(111-122) immunoreactivities eluted at the position of synthetic PHV. Surprisingly, a major peak of prepro-VIP(111-122) immunoreactivity eluted in a position not related to any other immunoreactivity indicating the presence of prepro-VIP(111-122). Measurements of these immunoreactivities in cortical and suprachiasmatic extracts revealed that VIP was found in the highest concentration whereas PHV was found in the lowest. Immunoreactivity for PHI and prepro-VIP(111-122) was found in moderate concentrations. Except for prepro-VIP(111-122) which was found to be approximately 3 x higher concentrated in the SCN than in the cerebral cortex, the other immunoreactivities were found in almost similar relative concentrations in the two tissues. Using immunocytochemistry, elongated neurons mostly of the bipolar type with prominent processes observed in the cerebral cortex reacted with all antisera tested. More PHI/PHV/prepro-VIP(111-122)- than VIP-immunoreactive (ir) nerve fibers were found in the cerebral cortex. In the SCN, the density of immunoreactivity was the same whatever antiserum used. VIP-, PHI- and prepro-VIP(111-122)/PHV-ir neurons were observed in the ventral part of the nucleus with numerous axons coursing caudodorsally into the subparaventricular area. A substantial number of terminals was detected caudal to the paraventricular nucleus. Minor projections spread to the medial part of the anterior nucleus and to the medial preoptic area hypothalamic. These data show that VIP and PHI are the major active peptides derived from prepro-VIP in the mouse cerebral cortex and SCN whereas PHV was found in minor concentrations. Prepro-VIP(111-122), which so far has been found to have no functional significance, is, therefore, most likely a vaste fragment of processing of PHI in central neurons.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Co-localisation of substance P-, bombesin- and peptide histidine isoleucine (PHI)-like peptides in gut endocrine cells of the dogfish Scyliorhinus stellaris.

The gastrointestinal tract of cartilaginous fishes, like that of higher vertebrates, is known to contain endocrine cells and nerves immunoreactive for a wide variety of peptides, some of which have been structurally characterised. Since we have found that substance P-, bombesin- and peptide histidine isoleucine-like immunoreactivities are similarly distributed in the endocrine cells of the dogfish pyloric stomach, we have tried to establish whether any of these peptides are co-localised. The cells were compared in thin serial sections with both light- and electron microscopical immunocytochemistry. Double immunolabelling was also used to show two immunoreactive peptides in the same tissue section. Further characterisation of the immunoreactivity was attempted by preabsorbing the antibodies with various peptides or synthetic fragments of peptide molecules. Immunoreactivity for all three peptides was frequently present in the same cells, whereas antibodies to other peptides such as gastrin and somatostatin marked different cells. Electron microscopy indicated that all the secretory granules in three morphologically different cell types reacted with antibodies to all three peptides. Dual localisation of unrelated peptides in endocrine cells or nerves is established in many cases, but triple localisation is as yet unusual. The immunoreaction for bombesin-like peptides is different in endocrine cells and nerves, indicating that dogfish bombesin may be present in two forms, in agreement with biochemical evidence.

Animals↗

Cyclic AMP formation in chicken brain: effect of vasoactive intestinal peptide, peptide histidine-isoleucine (PHI), and some PHI-related peptides.

Vasoactive intestinal peptide (chicken form; chVIP), peptide histidine-isoleucine (porcine and rat forms; pPHI and rPHI), D-Phe(4) derivative of porcine PHI (D-Phe(4)-pPHI), peptide histidine-methionine (PHM; human PHI), and helodermin, were tested for their ability to stimulate cAMP production in [(3)H]adenine-prelabeled slices of chick cerebral cortex (CCx) and hypothalamus (HTh). The chVIP (0.1-3 microM) concentration-dependently and potently stimulated cAMP production in HTh and CCx; the responses observed after 3 microM of chVIP were comparable to those produced by 0.1 microM PACAP38. Helodermin (5 microM) moderately but significantly stimulated cAMP formation in both HTh and CCx, whereas pPHI, rPHI, PHM at 5 microM concentration only weakly affected cAMP production in CCx, and were inactive in HTh; D-Phe(4)-pPHI was inactive in both tissues. These data demonstrate that chVIP, PACAP, and to a lesser extent helodermin were capable of potently stimulating cAMP generation in the avian central nervous system. PHI-related peptides showed only weak or no activity, depending on the tissue.

Animals↗