RESUMO
Energy balance is regulated, in part, by the orexigenic signaling pathways of the vagus nerve. Fasting-induced modifications in the expression of orexigenic signaling systems have been observed in vagal afferents of lean animals. Altered basal cannabinoid (CB1) receptor expression in the nodose ganglia in obesity has been reported. Whether altered body weight or a high fat diet modifies independent or additive changes in CB1 expression is unknown. We investigated the expression of CB1 and orexin 1 receptor (OX-1R) in the nodose ganglia of rats fed ad libitum or food deprived (24h), maintained on low or high fat diets (HFD), with differing body weights. Male Wistar rats were fed chow or HFD (diet-induced obese: DIO or diet-resistant: DR) or were body weight matched to the DR group but fed chow (wmDR). CB1 and OX-1R immunoreactivity were investigated and CB1 mRNA density was determined using in situ hybridization. CB1 immunoreactivity was measured in fasted rats after sulfated cholecystokinin octapeptide (CCK8s) administration. In chow rats, fasting did not modify the level of CB1 mRNA. More CB1 immunoreactive cells were measured in fed DIO, DR and wmDR rats than chow rats; levels increased after fasting in chow and wmDR rats but not in DIO or DR rats. In HFD fasted rats CCK8s did not reduce CB1 immunoreactivity. OX-1R immunoreactivity was modified by fasting only in DR rats. These data suggest that body weight contributes to the proportion of neurons expressing CB1 immunoreactivity in the nodose ganglion, while HFD blunts fasting-induced increases, and CCK-induced suppression of, CB1-immunoreactivity.
Assuntos
Peso Corporal/fisiologia , Dieta Hiperlipídica , Gânglio Nodoso/metabolismo , Receptor CB1 de Canabinoide/biossíntese , Animais , Ingestão de Energia , Imuno-Histoquímica , Hibridização In Situ , Masculino , Ratos , Ratos WistarRESUMO
BACKGROUND: In animal studies, gut vagal afferent neurons express cannabinoid (CB1) receptors, whose expression is increased by fasting. We aimed to explore the possibility that similar effects might be relevant in man in controlling gastric emptying. METHODS: Fourteen healthy volunteers underwent measurements of gastric emptying using the (13) C acetate breath test, after either a nutrient (skimmed milk) or non-nutrient (water) meal following both a 12 and 24 h fast. Further gastric emptying studies were performed with and without the CB1 receptor antagonist Rimonabant (20 mg or 80 mg). Because of the inter-individual variations observed, two subjects underwent additional studies with and without Rimonabant to determine intra-individual consistency. Gastric emptying was evaluated as cumulative C13 : C12 ratio values, measured at 5 min intervals for 30 min. KEY RESULTS: In the group as a whole, fasting duration slowed gastric emptying for both the nutrient [120 ± 30 (mean ± SD) vs 101 ± 34, P < 0.05] and non-nutrient [226 ± 62 vs 177 ± 47, P < 0.05] meals, but there was no effect of Rimonabant. However, there was consistent inter individual variation; thus while 12 subjects showed a slowing, two (14%) exhibited accelerated gastric emptying for both the nutrient and the non-nutrient meal after 24 h fasting and in one of whom, Rimonabant consistently reversed the fasting effect on the non-nutrient meal. CONCLUSIONS & INFERENCES: Extended fasting alters the gastric emptying of liquid meals but there are consistent differences between individuals. Where there is an accelerated response to fasting, Rimonabant appears to reverse the effect.
Assuntos
Jejum/fisiologia , Esvaziamento Gástrico/fisiologia , Antagonistas de Receptores de Canabinoides/farmacologia , Endocanabinoides , Esvaziamento Gástrico/efeitos dos fármacos , Humanos , Piperidinas/farmacologia , Pirazóis/farmacologia , RimonabantoRESUMO
The ingestion of food activates mechanisms leading to inhibition of food intake and gastric emptying mediated by the release of regulatory peptides, for example cholecystokinin (CCK), and lipid amides, e.g. oleylethanolamide from the gut. In addition, there are both peptides (e.g. ghrelin) and lipid amides (e.g. anandamide) that appear to signal the absence of food in the gut and that are associated with the stimulation of food intake. Vagal afferent neurones are a common target for both types of signal. Remarkably, the neurochemical phenotype of these neurones itself depends on nutritional status. CCK acting at CCK1 receptors on vagal afferent neurones stimulates expression in these neurones of Y2-receptors and the neuropeptide CART, both of which are associated with the inhibition of food intake. Conversely, in fasted rats when plasma CCK is low, these neurones express cannabinoid (CB)-1 and melanin concentrating hormone (MCH)-1 receptors, and MCH, and this is inhibited by exogenous CCK or endogenous CCK released by refeeding. The stimulation of CART expression by CCK is mediated by the activation of CREB and EGR1; ghrelin inhibits the action of CCK by promoting nuclear exclusion of CREB and leptin potentiates the action of CCK by the stimulation of EGR1 expression. Vagal afferent neurones therefore constitute a level of integration outside the CNS for nutrient-derived signals that control energy intake and that are capable of encoding recent nutrient ingestion.
Assuntos
Vias Aferentes/fisiologia , Regulação do Apetite/fisiologia , Jejum/fisiologia , Comportamento Alimentar/fisiologia , Plasticidade Neuronal/fisiologia , Células Receptoras Sensoriais/fisiologia , Nervo Vago/fisiologia , Animais , Humanos , Modelos Neurológicos , RatosRESUMO
Plasma concentrations of the hormone gastrin are elevated by Helicobacter pylori infection and by gastric atrophy. It has previously been proposed that gastrin acts as a cofactor during gastric carcinogenesis and hypergastrinemic transgenic INS-GAS mice are prone to developing gastric adenocarcinoma, particularly following H. pylori infection. We hypothesised that the increased risk of carcinogenesis in these animals may partly result from altered susceptibility of gastric epithelial cells to undergo apoptosis. Gastric corpus apoptosis was significantly increased 48 h after 12Gy gamma-radiation in mice rendered hypergastrinemic by transgenic (INS-GAS) or pharmacological (omeprazole treatment of FVB/N mice) methods and in both cases the effects were inhibited by the CCK-2 receptor antagonist YM022. However, no alteration in susceptibility to gamma-radiation-induced gastric epithelial apoptosis was observed in mice overexpressing progastrin or glycine-extended gastrin. Apoptosis was also significantly increased in gastric corpus biopsies obtained from H. pylori-infected humans with moderate degrees of hypergastrinemia. We conclude that hypergastrinemia specifically renders cells within the gastric corpus epithelium more susceptible to induction of apoptosis by radiation or H. pylori. Altered susceptibility to apoptosis may therefore be one factor predisposing to gastric carcinogenesis in INS-GAS mice and similar mechanisms may also be involved in humans.
Assuntos
Apoptose , Suscetibilidade a Doenças , Mucosa Gástrica/patologia , Gastrinas/sangue , Neoplasias Gástricas/etiologia , Animais , Antiulcerosos/farmacologia , Benzodiazepinas/farmacologia , Células Cultivadas , Raios gama , Mucosa Gástrica/efeitos dos fármacos , Mucosa Gástrica/efeitos da radiação , Helicobacter pylori , Humanos , Camundongos , Camundongos Transgênicos , Omeprazol/farmacologia , Receptor de Colecistocinina B/antagonistas & inibidoresRESUMO
BACKGROUND: Matrix metalloproteinases (MMPs) have roles in inflammation and other processes relevant to the architectural disturbances seen in the gastric mucosa in response to Helicobacter pylori infection. Upregulation of MMPs has been reported in H pylori infection, but there are no detailed reports regarding altered production of their inhibitors, the tissue inhibitors of metalloproteinases (TIMPs). AIMS: To investigate changes in the abundance of TIMPs in human gastric corpus mucosa and murine stomach in Helicobacter infection, and to study cellular sources in man. METHODS: Gastric corpus biopsy samples were assessed for abundance of mRNA or protein for TIMP-1 to -4 by real-time quantitative PCR or western blotting, respectively. Antral and corpus biopsies were processed for histology, H pylori status and inflammatory scoring. Cellular sources of TIMP-1, -3 and -4 were examined by indirect immunohistochemistry. Circulating gastrin was measured by radioimmunoassay. Also, abundance of TIMP-1, -3 and -4 mRNA in the stomach of Helicobacter felis infected mice post-infection was compared with that of uninfected control animals. RESULTS: Compared with uninfected patients, mRNA and protein for TIMP-1, -3 and -4 were significantly more abundant in the gastric corpus of H pylori infected subjects. Gastric TIMP expression did not differ significantly between hyper- and normogastrinaemic subjects within the H pylori negative and positive groups. There was no difference in mRNA abundance for MMP-3 or -8. Immunohistochemistry showed TIMP proteins localised to gastric epithelial, stromal cells and inflammatory cells. Murine H felis infection was associated with upregulation of TIMP-1 and -3 mRNA. CONCLUSIONS: Helicobacter infection is associated with upregulation of specific TIMPs (TIMP-1 and -3) in glandular epithelium and stroma. It is suggested that increased expression of specific protease inhibitors in the corpus mucosa may exert important effects on extracellular matrix remodelling and influence the outcome of H pylori infection.
Assuntos
Mucosa Gástrica/metabolismo , Gastrite/metabolismo , Infecções por Helicobacter/metabolismo , Helicobacter pylori , Inibidores Teciduais de Metaloproteinases/metabolismo , Animais , Gastrinas/sangue , Gastrite/sangue , Gastrite/microbiologia , Gastrite/patologia , Infecções por Helicobacter/sangue , Infecções por Helicobacter/patologia , Humanos , Camundongos , Camundongos Transgênicos , RNA Mensageiro/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Índice de Gravidade de Doença , Inibidores Teciduais de Metaloproteinases/genética , Regulação para CimaRESUMO
Food intake is regulated by signals from the gastrointestinal tract. Both stimulation and inhibition of food intake may be mediated by upper gastrointestinal tract hormones, e.g. ghrelin and cholecystokinin that act at least partly via vagal afferent neurones. We now report that vagal afferent neurones in both rat and man express melanin-concentrating hormone and its receptor, melanin-concentrating hormone-1R. In nodose ganglia from rats fasted for 24 h, RT-PCR revealed the expression of both melanin-concentrating hormone and melanin-concentrating hormone-1R, whereas in ganglia from animals fed ad libitum expression was virtually undetectable. Immunohistochemical studies also revealed expression of melanin-concentrating hormone and melanin-concentrating hormone-1R in nodose ganglion neurones of fasted rats, but signals were weak in rats fed ad libitum. Melanin-concentrating hormone and melanin-concentrating hormone-1R were expressed in the same neurones, a high proportion of which also expressed the cholecystokinin-1 receptor. When fasted rats were refed, there was down-regulation of melanin-concentrating hormone and melanin-concentrating hormone-1R expression over a period of 5 h. Similar effects were produced by administration of cholecystokinin to fasted rats. The cholecystokinin-1 receptor antagonist lorglumide inhibited food-induced down-regulation of melanin-concentrating hormone and melanin-concentrating hormone-1R. We conclude that the satiety hormone cholecystokinin acts on vagal afferent neurones to inhibit expression of melanin-concentrating hormone and its receptor. Since the melanin-concentrating hormone system is associated with stimulation of food intake this effect of cholecystokinin may contribute to its action as a satiety hormone.
Assuntos
Vias Aferentes/fisiologia , Hormônios Hipotalâmicos/fisiologia , Melaninas/fisiologia , Neurônios/fisiologia , Hormônios Hipofisários/fisiologia , Receptores de Somatostatina/fisiologia , Nervo Vago/fisiologia , Animais , Jejum , Comportamento Alimentar , Masculino , Gânglio Nodoso/fisiologia , Ratos , Ratos Wistar , Resposta de SaciedadeRESUMO
Transgenic mice (hGAS) that overexpress human progastrin are more susceptible than wild-type mice (FVB/N) to the induction of colonic aberrant crypt foci (ACF) and adenomas by the chemical carcinogen azoxymethane. We have previously shown significantly increased levels of colonic mitosis in hGAS compared with FVB/N mice after gamma-radiation. To investigate whether the effects of progastrin observed in hGAS colon require the presence of other forms of circulating gastrin, we have crossed hGAS (hg(+/+)) with gastrin knockout (G(-/-)) mice to generate mice that express progastrin and no murine gastrin (G(-/-)hg(+/+)). After azoxymethane, G(-/-)hg(+/+) mice developed significantly more ACF than control G(-/-)hg(-/-) mice (which do not express any forms of gastrin). G(-/-)hg(+/+) mice also exhibited significantly increased colonic mitosis both before and after exposure to 8 Gray Gy gamma-radiation or 50 mg/kg azoxymethane compared with G(-/-)hg(-/-). Treatment of G(-/-)hg(-/-) mice with synthetic progastrin (residues 21-101 of human preprogastrin) or G17 extended at its COOH terminus corresponding to the COOH-terminal 26-amino-acid residues of human preprogastrin (residues 76-101, G17-CFP) resulted in continued colonic epithelial mitosis after gamma-radiation, whereas glycine-extended gastrin-17 and the COOH-terminal tryptic fragment of progastrin [human preprogastrin-(96-101)] had no effect. Immunoneutralization with an antibody against G17-CFP before gamma-radiation significantly decreased colonic mitosis in G(-/-)hg(+/+) mice to levels similar to G(-/-)hg(-/-). We conclude that progastrin does not require the presence of other forms of gastrin to exert proliferative effects on colonic epithelia and that the portion of the peptide responsible for these effects is contained within amino acid residues 76-101 of human preprogastrin.
Assuntos
Colo/citologia , Células Epiteliais/efeitos dos fármacos , Gastrinas/farmacologia , Mitógenos , Mitose/efeitos dos fármacos , Fragmentos de Peptídeos/farmacologia , Precursores de Proteínas/farmacologia , Sequência de Aminoácidos , Animais , Antimetabólitos , Apoptose/efeitos dos fármacos , Azoximetano/farmacologia , Bromodesoxiuridina , Carcinógenos/farmacologia , Colo/efeitos dos fármacos , Dano ao DNA/efeitos dos fármacos , Dano ao DNA/efeitos da radiação , Raios gama , Gastrinas/química , Gastrinas/genética , Genótipo , Humanos , Imuno-Histoquímica , Camundongos , Camundongos Knockout , Camundongos Transgênicos , Fragmentos de Peptídeos/química , Precursores de Proteínas/químicaAssuntos
Colecistocinina/sangue , Giardíase/sangue , Doença Aguda , Adulto , Biomarcadores/sangue , Feminino , Humanos , Masculino , Pessoa de Meia-IdadeAssuntos
Carcinoma Neuroendócrino/patologia , Proteínas de Ligação a DNA/fisiologia , Neoplasias Gastrointestinais/patologia , Proteínas de Neoplasias/fisiologia , Neoplasias Pancreáticas/patologia , Proteínas Proto-Oncogênicas , Transativadores/fisiologia , Fator de Crescimento Transformador beta/fisiologia , Animais , Humanos , Ratos , Receptores de Fatores de Crescimento Transformadores beta/fisiologia , Proteínas SmadRESUMO
The wall of the gut responds to an impressive array of signals originating in the lumen, including nutrient and non-nutrient chemicals, mechanical factors, and micro-organisms. The idea that the gut wall exhibits luminal chemo-sensitivity is implied in the original discovery of secretin by Bayliss and Starling, and has become an integral part of models of neurohumoral control of gastrointestinal function. Entero-endocrine cells are specialised for luminal nutrient sensing but sub-epithelial nerve fibres may also respond to luminal chemicals that freely diffuse across the epithelium eg short chain fatty acids. The molecular recognition mechanisms include G-protein coupled receptors (GPCRs) eg the extracellular Ca(2+) sensing receptor which also responds to aromatic amino acids. There are also GPCRs sensing fatty acids, as well as bitter or noxious compounds. In addition, though, gating of ion channels including events secondary to energy availability eg ATP, may be involved in sensing some luminal chemicals. There is likely to be integration of luminal signals at several levels including at the level of entero-endocrine cells and at sub-epithelial nerve fibers. For example, the intestinal hormone CCK acts on primary afferent nerve fibers of the vagal trunk. The same fibers also express leptin receptors that are thought to respond to leptin released from gastric chief cells, orexin receptors (activation of which inhibits CCK) and possibly ghrelin receptors. Multiple signalling mechanisms allow specific responses to be matched to meals of differing content.
Assuntos
Células Enteroendócrinas/fisiologia , Trato Gastrointestinal/fisiologia , Animais , Ingestão de Alimentos/fisiologia , Células Enteroendócrinas/citologia , Células Enteroendócrinas/metabolismo , Trato Gastrointestinal/citologia , Trato Gastrointestinal/inervação , Trato Gastrointestinal/metabolismo , HumanosRESUMO
Recent studies suggest that gastrin regulates parietal cell maturation. We asked whether it also regulates parietal cell life span and migration along the gland. Dividing cells were labeled with 5'-bromo-2'-deoxyuridine (BrdU), and parietal cells were identified by staining with Dolichos biflorus lectin. Cells positive for D. biflorus lectin and BrdU were reliably identified 10-30 days after BrdU injection in mice in which the gastrin gene had been deleted by homologous recombination (Gas-KO) and wild-type (C57BL/6) mice. The time course of labeling was similar in the two groups. The distribution of BrdU-labeled parietal cells in wild-type mice was consistent with migration to the base of the gland, but in Gas-KO mice, a higher proportion of BrdU-labeled cells was found more superficially 20 and 30 days after BrdU injection. Conversely, in transgenic mice overexpressing gastrin, BrdU-labeled parietal cells accounted for a higher proportion of the labeled pool in the base of the gland 10 days after BrdU injection. Gastrin, therefore, stimulates movement of parietal cells along the gland axis but does not influence their life span.
Assuntos
Gastrinas/fisiologia , Células Parietais Gástricas/fisiologia , Lectinas de Plantas , Animais , Antibacterianos/farmacologia , Bromodesoxiuridina , Movimento Celular/fisiologia , Sobrevivência Celular , Gastrinas/sangue , Gastrinas/genética , ATPase Trocadora de Hidrogênio-Potássio/metabolismo , Lectinas/farmacocinética , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout/genética , Células Parietais Gástricas/efeitos dos fármacos , Distribuição TecidualRESUMO
Epithelial organization is maintained by cell proliferation, migration, and differentiation. In the case of the gastric epithelium, at least some of these events are regulated by the hormone gastrin. In addition, gastric epithelial cells are organized into characteristic tubular structures (the gastric glands), but the cellular mechanisms regulating the organization of tubular structures (sometimes called branching morphogenesis) are uncertain. In the present study, we examined the role of the gastrin-cholecystokinin(B) receptor in promoting branching morphogenesis of gastric epithelial cells. When gastric cancer AGS-G(R) cells were cultured on plastic, gastrin and PMA stimulated cell adhesion, formation of lamellipodia, and extension of long processes in part by activation of protein kinase C (PKC) and phosphatidylinositol (PI)-3 kinase. Branching morphogenesis was not observed in these circumstances. However, when cells were cultured on artificial basement membrane, the same stimuli increased the formation of organized multicellular arrays, exhibiting branching morphogenesis. These effects were reversed by inhibitors of PKC but not of PI-3 kinase. We conclude that, in the presence of basement membrane, activation of PKC by gastrin stimulates branching morphogenesis.
Assuntos
Receptores da Colecistocinina/metabolismo , Neoplasias Gástricas/patologia , Membrana Basal/patologia , Adesão Celular/efeitos dos fármacos , Gastrinas/farmacologia , Humanos , Lisofosfolipídeos/farmacologia , Proteína Quinase C/fisiologia , Receptor de Colecistocinina B , Neoplasias Gástricas/fisiopatologia , Acetato de Tetradecanoilforbol/farmacologia , Células Tumorais Cultivadas/efeitos dos fármacosRESUMO
BACKGROUND: Activation of the gastrin-cholecystokinin(B) (CCK(B)) receptor stimulates cell proliferation and increases production of ligands for the epidermal growth factor receptor (EGF-R). AIMS: To determine the role of gastrin-CCK(B) activation in stimulation of cell proliferation via paracrine activation of EGF-R. METHODS: AGS cells were transfected with the gastrin-CCK(B) receptor (AGS-G(R) cells) or with green fluorescent protein (AGS-GFP cells). Proliferation was determined by [(3)H] thymidine incorporation, flow cytometry, and cell counting. RESULTS: Gastrin inhibited proliferation of AGS-G(R) cells by delaying entry into S phase. However, when AGS-G(R) cells were cocultured with AGS-GFP cells, gastrin stimulated proliferation of the latter. Immunoneutralisation and pharmacological studies using metalloproteinase and kinase inhibitors indicated that the proliferative response was mediated by paracrine stimulation of EGF-R and activation of the mitogen activated protein kinase pathway through release of heparin binding EGF. CONCLUSIONS: Gastrin can directly inhibit, and indirectly stimulate, proliferation of gastric AGS cells.
Assuntos
Células Enterocromafins/metabolismo , Receptores ErbB/metabolismo , Receptores da Colecistocinina/metabolismo , Divisão Celular/fisiologia , Citometria de Fluxo , Proteínas de Fluorescência Verde , Humanos , Proteínas Luminescentes/metabolismo , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Receptor de Colecistocinina B , Timidina/metabolismoRESUMO
There is evidence for interactions between leptin and cholecystokinin in controlling food intake. Since cholecystokinin acts on vagal afferent neurones, we asked whether the leptin receptor was also expressed by these neurones. Primers for different forms of the leptin receptor were used in reverse transcriptase-polymerase chain reaction (RT-PCR) of rat and human nodose ganglia. RT-PCR yielded products corresponding to the long (functional) form as well as short forms of the rat leptin receptor. Moreover, RT-PCR revealed the long form of the leptin receptor in a human nodose ganglion. The identities of RT-PCR products were confirmed by sequencing. Primers corresponding to leptin itself did not give RT-PCR products in nodose ganglia. Immunocytochemical studies revealed leptin-receptor immunoreactivity in neuronal cell bodies. Many neurones co-expressed the leptin and cholecystokinin type A receptors, or leptin receptor and cocaine- and amphetamine-related transcript. We conclude that vagal afferent neurones that express the cholecystokinin type A receptor and cocaine- and amphetamine-related transcript, may also express the long form of the leptin receptor providing a neurochemical basis for observations of interactions between cholecystokinin and leptin.
Assuntos
Regulação do Apetite/fisiologia , Proteínas de Transporte/genética , Leptina/metabolismo , Neurônios Aferentes/metabolismo , Gânglio Nodoso/metabolismo , Receptores de Superfície Celular , Fibras Aferentes Viscerais/metabolismo , Animais , Peptídeo Relacionado com Gene de Calcitonina/genética , Peptídeo Relacionado com Gene de Calcitonina/metabolismo , Proteínas de Transporte/metabolismo , Colecistocinina/metabolismo , DNA Complementar/genética , Imunofluorescência , Humanos , Hibridização In Situ , Microscopia Confocal , Dados de Sequência Molecular , Proteínas do Tecido Nervoso/genética , Proteínas do Tecido Nervoso/metabolismo , Neurônios Aferentes/citologia , Gânglio Nodoso/citologia , RNA Mensageiro/metabolismo , Ratos , Ratos Wistar , Receptor de Colecistocinina A , Receptores da Colecistocinina/genética , Receptores da Colecistocinina/metabolismo , Receptores para Leptina , Homologia de Sequência do Ácido Nucleico , Fibras Aferentes Viscerais/citologiaRESUMO
1. The acidic interior of neuroendocrine secretory vesicles provides both an energy gradient for amine-proton exchangers (VMATs) to concentrate small transmitter molecules, for example catecholamines, and an optimal pH for the prohormone convertases which cleave hormone precursors. There is evidence that VMAT activity modulates prohormone cleavage, but in the absence of measurements of pH in secretory vesicles in intact cells, it has not been possible to establish whether these effects are attributable to raised intravesicular pH due to proton transport through VMATs. 2. Clones were generated of the hamster insulinoma cell line HIT-T15 expressing a pH-sensitive form of green fluorescent protein (GFP-F64L/S65T) targeted to secretory vesicles, with and without co-expression of VMAT2. In order to study prohormone cleavage, further clones were generated that expressed preprogastrin with and without co-expression of VMAT2. 3. Confocal microscopy of GFP fluorescence indicated that the pH in the secretory vesicles was 5.6 in control cells, compared with 6.6 in cells expressing VMAT2; the latter was reduced to 5.8 by the VMAT inhibitor reserpine. 4. Using a pulse-chase labelling protocol, cleavage of 34-residue gastrin (G34) was found to be inhibited by co-expression with VMAT2, and this was reversed by reserpine. Similar effects on vesicle pH and G34 cleavage were produced by ammonium chloride. 5. We conclude that VMAT expression confers the linked abilities to store biogenic amines and modulate secretory vesicle pH over a range influencing prohormone cleavage and therefore determining the identity of regulatory peptide secretory products.
Assuntos
Álcalis/metabolismo , Hidrogênio/metabolismo , Glicoproteínas de Membrana/metabolismo , Proteínas de Membrana Transportadoras , Neuropeptídeos , Sistemas Neurossecretores/metabolismo , Vesículas Secretórias/metabolismo , Cloreto de Amônio/farmacologia , Animais , Cricetinae , Gastrinas/antagonistas & inibidores , Gastrinas/efeitos dos fármacos , Gastrinas/metabolismo , Gastrinas/fisiologia , Proteínas de Fluorescência Verde , Concentração de Íons de Hidrogênio , Indicadores e Reagentes , Proteínas Luminescentes , Glicoproteínas de Membrana/antagonistas & inibidores , Microscopia Confocal , Sistemas Neurossecretores/citologia , Precursores de Proteínas/antagonistas & inibidores , Precursores de Proteínas/efeitos dos fármacos , Precursores de Proteínas/metabolismo , Precursores de Proteínas/fisiologia , Processamento de Proteína Pós-Traducional/efeitos dos fármacos , Reserpina/farmacologia , Células Tumorais Cultivadas , Proteínas Vesiculares de Transporte de Aminas Biogênicas , Proteínas Vesiculares de Transporte de MonoaminaRESUMO
Gastric epithelial organization and function are controlled and maintained by a variety of endocrine and paracrine mediators. Peptides encoded by the gastrin gene are an important part of this system because targeted deletion of the gene, or of the gastrin-CCKB receptor gene, leads to decreased numbers of parietal cells and decreased gastric acid secretion. Recent studies indicate that the gastrin precursor, preprogastrin, gives rise to a variety of products, each with a distinctive spectrum of biological activity. The conversion of progastrin to smaller peptides is regulated by multiple mechanisms including prohormone phosphorylation and secretory vesicle pH. Progastrin itself stimulates colonic epithelial proliferation; biosynthetic intermediates (Gly-gastrins) stimulate colonic epithelial proliferation and gastric epithelial differentiation; and C-terminally amidated gastrins stimulate colonic proliferation, gastric epithelial proliferation and differentiation, and acid secretion. The effects of progastrin-derived peptides on gastric epithelial function are mediated in part by release of paracrine factors that include histamine, epidermal growth factor (EGF)-receptor ligands, and Reg. The importance of the appropriate regulation of this system is shown by the observation that prolonged moderate hypergastrinemia in transgenic mice leads to remodelling of the gastric epithelium, and in the presence of Helicobacter, to gastric cancer.
Assuntos
Gastrinas/biossíntese , Gastrinas/metabolismo , Células Parietais Gástricas/metabolismo , Animais , HumanosRESUMO
BACKGROUND & AIMS: Studies in gastrin-deficient mice have demonstrated critical roles for gastrin peptides in the regulation of gastric acid secretion, but the relative contributions of amidated (G-17) and glycine-extended (G17-Gly) gastrin remain unclear. We examined the effects of these 2 forms of gastrin on acid secretion in gastrin-deficient mice. METHODS: Sixty gastrin-deficient mice received infusions of saline, or 1, 6, or 14 days of amidated gastrin 17 (G-17), G17-Gly, or both G-17 and G17-Gly at 10 nmol. kg(-1). h(-1). Twenty-four gastrin-deficient mice were then infused for 14 days with 1, 2, or 5 nmol. kg(-1). h(-1) of G-17 or G-17 and G17-Gly. Acid secretion was determined 4 hours after pyloric ligation, and gastric tissue was processed for histology, immunohistochemistry, and electron microscopy. RESULTS: Infusion of G-17 increased acid secretion in a dose-dependent manner with a peak at 5 nmol. kg(-1). h(-1) and a subsequent decrease in acid secretion at higher doses. Infusion of G17-Gly alone had no effect on acid secretion, but coinfusion with G-17 resulted in significantly higher levels of acid secretion at all doses examined than infusion with G-17 alone. The potentiating effect of G17-Gly on G-17-induced acid secretion was associated with increased parietal cell activation but was independent of changes in parietal and enterochromaffin-like cell number, fundic proliferation rates, and H(+),K(+)-adenine triphosphatase expression. G17-Gly also prevented the formation of vacuolar canaliculi and lipofuscin bodies in the parietal cells induced by G-17. CONCLUSIONS: G17-Gly appears to synergize with G-17 to up-regulate acid secretion and prevent parietal cell degradation. These results suggest that G17-Gly plays an important role in parietal cell function.