Regulatory Peptides in the Mammalian Respiratory Tract*

Ghatei, Mohammad A.; Sheppard, Mary N.; O’Shaughnessy, Denis; Adrian, Thomas E.; McGregor, G.P.; Polak, Julia M.; Bloom, Stephen R.

doi:10.1210/endo-111-4-1248

Cited by 123 publications

(32 citation statements)

References 18 publications

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“…125I-GRP binding was reduced to 5% of total cell-associated radioactivity by unlabeled GRP (1 ,ug/ml); in contrast, none of the other factors tested caused a significant decrease in 125I-GRP binding, even at this low concentration of labeled Table 2) enabled us to examine the structure-activity relationships for their ability both to inhibit 125I-GRP binding and to stimulate DNA synthesis. The addition of unlabeled GRP, GRP-(14-27), neuromedin B, or the 8-to 14-amino acid fragment of bombesin [bombesin- (8)(9)(10)(11)(12)(13)(14)] to 3T3 cells inhibited specific 125I-GRP binding in a concentration-dependent manner (Fig. 2 Lower).…”

Section: I-grp Binds To Specific High-affiity Sites In Swiss 3t3mentioning

confidence: 88%

“…2 Lower). The Ki values for GRP, GRP-(14-27), neuromedin B, and bombesin- (8)(9)(10)(11)(12)(13)(14) were, respectively, 0.7, 0.33, 7.6, and 888 x 10-9 M. In contrast, the NH2-terminal fragment of GRP [GRP-(1-16)] did not inhibit binding at concentrations up to 5 ,g/ml. It is noteworthy that 1251I-GRP stimulated DNA synthesis in Swiss 3T3 cells with the same potency as did unlabeled GRP.…”

Section: I-grp Binds To Specific High-affiity Sites In Swiss 3t3mentioning

confidence: 99%

“…Peptides related to bombesin, including gastrin-releasing peptide (GRP) and the neuromedins, are widely distributed in normal mammalian tissues (10)(11)(12)(13)(14)(15)(16) and also are found in high levels in human pulmonary (17)(18)(19)(20) and thyroid (21) tumors. The finding that bombesin is mitogenic (9) and the wide distribution of endogenous mammalian analogues of bombesin strongly suggest that this family of peptides may play a role in the control of both normal and abnormal animal cell growth.…”

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High-affinity receptors for peptides of the bombesin family in Swiss 3T3 cells.

Zachary¹,

Rozengurt²

1985

Proc. Natl. Acad. Sci. U.S.A.

178

134

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Gastrin-releasing peptide (GRP) Peptides related to bombesin, including gastrin-releasing peptide (GRP) and the neuromedins, are widely distributed in normal mammalian tissues (10-16) and also are found in high levels in human pulmonary (17)(18)(19)(20) and thyroid (21) tumors. The finding that bombesin is mitogenic (9) and the wide distribution of endogenous mammalian analogues of bombesin strongly suggest that this family of peptides may play a role in the control of both normal and abnormal animal cell growth.An important step in the elucidation of the mechanism of bombesin-stimulated mitogenesis is to determine the presence of specific receptors for this peptide in Swiss 3T3 cells. We have used biologically active, radiolabeled GRP, the mammalian counterpart of bombesin, to establish the presence of receptors for this molecule.In the present paper, we show that the mitogenic stimulation of Swiss 3T3 cells by peptides of the bombesin family is mediated by a single class of high-affinity receptors. This receptor is distinct from those of other mitogens for these cells. Further, a substance P (SP) analogue recently described as a bombesin antagonist (22) selectively blocks both GRP binding and GRP-induced mitogenesis. Since there is evidence that bombesin-like peptides may contribute to the self-stimulatory (autocrine) growth of small-cell carcinoma of the lung (18-20, 23, 24, 47), our findings may suggest novel clinical approaches to the treatment of this important cancer. MATERIALS AND METHODSCell culture procedures (9,25,26) and assays of DNA synthesis by [3H]thymidine incorporation (26) were carried out as described.Binding Assay. For binding at 37°C, confluent and quiescent cultures of Swiss 3T3 cells in 33-mm dishes were washed twice with Dulbecco's modified Eagle's medium (DME medium) and incubated with 1 ml of binding medium, which consisted of 1:1 (vol/vol) DME and Waymouth media supplemented with 1 mg of bovine serum albumin per ml, 50 mM 2-[bis(2-hydroxyethyl)-2-amino]ethanesulfonic acid (Bes) (pH 7.0), and GRP labeled with 125I at tyrosine-15 at the concentrations indicated. After 30 min of incubation (unless otherwise stated), cultures were washed rapidly four times with cold (4°C) wash solution (0.15 M NaCl/5 mM KCl/0.02 M Na2HPO4/1.8 mM KH2PO4, pH 7.2, supplemented with bovine serum albumin at 1 mg/ml). Washed cultures were extracted in 0.5 ml of 0.1 M NaOH containing 2% Na2CO3 and 1% NaDodSO4, and total cell-associated radioactivity was determined in a y counter. Nonspecific binding, defined as the cell-associated radioactivity not displaced in the presence of 360 nM GRP, was proportional to the concentration of 1251-GRP and varied from 23% of total binding at the highest concentrations (3 nM) of 125I-GRP to 5% at low concentrations (0.05 nM). For binding at 4°C, cells were washed once with DME medium at 4°C and incubated at 4°C for 5 min prior to the binding assay. The binding assay at 4°C was performed in 0.14 M NaCl/5 mM KCl/0.01 M Na2-HPO4/1.8 mM KH2PO4/1.8 mM CaCl2/1 mM MgCl2/25 ...

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Section: I-grp Binds To Specific High-affiity Sites In Swiss 3t3mentioning

confidence: 88%

Section: I-grp Binds To Specific High-affiity Sites In Swiss 3t3mentioning

confidence: 99%

mentioning

confidence: 99%

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High-affinity receptors for peptides of the bombesin family in Swiss 3T3 cells.

Zachary¹,

Rozengurt²

1985

Proc. Natl. Acad. Sci. U.S.A.

178

134

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“…The specificity of VIP antiserum was confirmed by lack of staining in adjacent sections incubated with antiserum pre-absorbed with VIP soluble antigen in the dilution range 0.01-0.1 mmol ml-' diluted antiserum. The antiserum did not show cross reactivity with other peptides known to be present in the lung (Bishop et al, 1980;Ghatei et al, 1982).…”

Section: Immunocytochemistrymentioning

confidence: 95%

Vasoactive intestinal peptide in bovine pulmonary artery: localisation, function and receptor autoradiography

Barnes

Cadieux

Carstairs

et al. 1986

British J Pharmacology

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1 The role of vasoactive intestinal peptide (VIP) in the control of pulmonary vascular tone was investigated by functional response, immunocytochemical localisation and receptor autoradiography in bovine pulmonary arteries. 2 VIP-immunoreactive nerve fibres were present at the adventitial-medial junction and in the media of the vessels. 3 Exposure of precontracted bovine pulmonary artery segments to VIP in vitro resulted in almost complete (86 ± 3%; mean ± s.e.mean) relaxation, the concentration needed for 50% relaxation being 4.47 ± 0.37 x 10-9M. VIP effects did not depend on the presence of intact endothelial cells. 4 The distribution of VIP receptors was studied by autoradiography using ['251I]-VIP. A high density of VIP receptors was found in arterial vascular smooth muscle, with a gradient of density from adventitia to luminal surface. There were no receptors on endothelial cells. 5 These data show that VIP is a potent vasodilator ofbovine pulmonary arteries, via direct activation of VIP receptors in vascular smooth muscle. VIP-immunoreactive nerves may influence pulmonary vascular tone directly and could, therefore, be important in regulating pulmonary blood flow.

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“…Substance P is present in sensory nerves in the airDr. Kohrogi ways of several species (6), including humans (7). Substance P stimulates many airway responses including gland secretion (8,9), smooth muscle contraction (10,11), increased vascular permeability (12), and neutrophil adhesion to the venous endothelium ( 13), and it potentiates cholinergic neurotransmission (1 1, 14).…”

Section: Introductionmentioning

confidence: 99%

Recombinant human enkephalinase (neutral endopeptidase) prevents cough induced by tachykinins in awake guinea pigs.

et al. 1989

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To determine whether recombinant enkephalinase (neutral endopeptidase, EC 3.4.24.11) prevents cough induced by exogenously applied and endogenously released neuropeptides, we measured cough responses to aerosolized solutions of substance P or of capsaicin for 2 min in random-source guinea pigs before or after exposing them to aerosolized recombinant human enkephalinase. Substance P (10-16 M) increased coughing compared with its vehicle. Enkephalinase (120 pg) inhibited cough induced by subsequent exposure to substance P compared with the response to substance P alone, but after further exposure to the enkephalinase inhibitor leucine-thiorphan (10-5 M), substance P increased cough significantly. Similar results were obtained for capsaicin-induced cough. In pathogen-free guinea pigs, after they inhaled inactive recombinant enkephalinase (33 ,ug), capsaicin (10-13 M) increased cough significantly. In contrast, after they inhaled active recombinant enkephalinase (33 ,ug), capsaicin increased cough only slightly. These results suggest that aerosolized enkephalinase reaches the sites of release or actions of endogenous neuropeptides and, by degrading them, prevents cough induced by their release. Furthermore, these studies suggest that recombinant enkephalinase might be useful in the treatment of cough and other symptoms of diseases involving peptides cleaved by this enzyme.

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Regulatory Peptides in the Mammalian Respiratory Tract*

Cited by 123 publications

References 18 publications

High-affinity receptors for peptides of the bombesin family in Swiss 3T3 cells.

High-affinity receptors for peptides of the bombesin family in Swiss 3T3 cells.

Vasoactive intestinal peptide in bovine pulmonary artery: localisation, function and receptor autoradiography

Recombinant human enkephalinase (neutral endopeptidase) prevents cough induced by tachykinins in awake guinea pigs.

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