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Biomedical subjects

J C Prieto

Publications and source records attributed to J C Prieto.

At least 109 records · Page 6Linked to original sources

Interaction of insulin with small intestinal epithelial cells from developing rats.

The interaction of insulin with its specific receptors was studied in jejunoileal epithelial cells from 14-, 20-, and 60-day-old rats. Characteristics such as potency and specificity of the insulin receptor system did not vary during development. However, insulin binding decreased with age due to a diminished concentration of insulin receptors, whereas the circulating levels of the hormone followed an inverse evolution. These results support previous suggestions on the potential role of insulin on the mechanisms of differentiation and proliferation of small intestinal mucosa.

Animals↗

The vasoactive intestinal peptide receptor-effector system is developmentally regulated in rat jejuno-ileal epithelial cells.

The specific binding of vasoactive intestinal peptide (VIP) to its specific receptors as well as the stimulatory effect of the neuropeptide on cyclic AMP accumulation were studied in jejuno-ileal epithelial cells from 14-, 20- and 60-day-old rats. The potency and specificity of the VIP receptor-effector system did not vary during development. However, the concentration of VIP receptors and the efficiency of VIP stimulation of cyclic AMP generation increased from suckling to adult conditions, and VIP levels in jejuno-ileal tissue followed a parallel course.

Aging↗

Inhibitory action of tumor-promoting phorbol esters upon beta-adrenergic agonist stimulation of cyclic AMP accumulation in rat prostatic epithelial cells.

The pretreatment of rat prostatic epithelial cells with 4 beta-phorbol 12-myristate 13-acetate resulted in an attenuation of beta-adrenergic stimulated cyclic AMP accumulation. The effect was dependent on time and concentration. The maximal extent of isoproterenol stimulation of cyclic AMP production was reduced by 35% after 15-min pretreatment with the phorbol ester at 25 degrees C. Since a similar action was exerted by other protein kinase C stimulators, present results suggest the involvement of this enzyme in a process of desensitization to beta-adrenergic agonists of the adenylate cyclase system in rat prostatic epithelium.

Adrenergic beta-Agonists↗

Tumor-promoting phorbol esters interfere with the vasoactive intestinal peptide receptor/effector system in rat prostatic epithelial cells.

Pretreatment of rat prostatic epithelial cells with the tumor-promoting phorbol ester 4 beta-phorbol 12-myristate 13-acetate resulted in a decrease of both the potency of vasoactive intestinal peptide (VIP) upon the stimulation of cyclic AMP accumulation and the affinity of the receptors of this peptide. These effects were dose-dependent and could be reproduced by other stimulators of protein kinase C (PKC). Thus, it is conceivably that phosphorylation of VIP receptors by PKC regulates VIP receptor function in the prostate gland.

Animals↗

Somatostatin inhibits VIP- and isoproterenol-stimulated cyclic AMP accumulation in rat prostatic epithelial cells.

The dual regulation of cyclic AMP accumulation was studied in rat prostatic epithelial cells incubated with somatostatin, vasoactive intestinal peptide (VIP), and the beta-adrenergic agent isoproterenol. Somatostatin noncompetitively inhibited the stimulatory effect of VIP and isoproterenol, but it did not alter basal cyclic AMP levels. In addition to the multifactorial regulation of the cyclic AMP system in rat prostatic epithelium, these results suggest that somatostatin may play a physiological role at this level.

Adenylyl Cyclase Inhibitors↗

Growth hormone binding and stimulation of amino acid uptake in epithelial cells of rat ventral prostate.

The binding of [125I]-human growth hormone (hGH) was studied in epithelial cells isolated from rat ventral prostate. Binding and degradation were dependent on time and temperature. The effect of a lysosomotropic agent suggested internalization and lysosomal degradation of the hormone. Dissociation and stoichiometric studies indicated the existence of a single class of GH receptors with a Kd of 0.7 nM and a binding capacity of 46 fmol hGH bound mg-1 cell protein. The receptor appeared to possess a somatotrophic nature since lactogenic hormones such as human placental lactogen and rat prolactin exhibited a very low degree of competition (whereas a variety of unrelated hormones and neuropeptides showed no effect). GH-stimulated leucine uptake by the cells in a time- and dose-dependent manner, half maximal effect being observed at 0.32 nM GH thus suggesting a direct relationship with the binding step.

Amino Acids↗

Characterization and age dependence of the stimulatory effect of VIP on cyclic AMP accumulation in rat Leydig cells.

The neuropeptide vasoactive intestinal peptide (VIP) has been shown to stimulate cyclic AMP accumulation in Leydig cells isolated from rat testis. The effect was dependent on time, temperature and cell concentration. At 15 degrees C, half-maximal and maximal stimulation were observed at about 1 and 100 nM VIP, respectively. The interaction was specific since an order of potencies chicken VIP greater than rat VIP greater than secretin greater than glucagon and no effect of neurotensin and substance P were obtained. The efficiency of VIP was lower in pubertal rats and then increased in young-adult and adult animals. These results together with the known presence of VIP in the testis support the idea that VIP may be involved in the regulation and function of Leydig cells during development.

Aging↗

Effect of cysteamine on cytosolic somatostatin binding sites in rabbit duodenal mucosa.

Administration of cysteamine in rabbits elicited a rapid depletion of both duodenal mucosa and plasma somatostatin. A significant reduction was observed within 5 min, returning toward control values by 150 min. The depletion of somatostatin was associated with an increase in the binding capacity and a decrease in the affinity of both high- and low-affinity binding sites present in cytosol of duodenal mucosa. Incubation of cytosolic fraction from control rabbits with 1 mM cysteamine did not modify somatostatin binding. Furthermore, addition of cysteamine at the time of binding assay did not affect the integrity of 125I-Tyr11-somatostatin. It is concluded that in vivo administration of cysteamine to rabbits depletes both duodenal mucosa and plasma somatostatin and leads to up-regulation of duodenal somatostatin binding sites.

Animals↗

Characterization of specific somatostatin binding sites in guinea pig lung.

Somatostatin binding sites have been demonstrated in the cytosolic fraction of guinea-pig lung. Binding of 125I-Tyr11-somatostatin was dependent on time and temperature, saturable, reversible and highly specific. Under equilibrium condition, i.e. 60 min at 25 degrees C, native somatostatin inhibited tracer binding in a dose-dependent manner. Two types of somatostatin binding sites were defined by Scatchard analysis: a small population with a high affinity (Kd = 23.4 nM) and a large population with a low affinity (Kd = 253.5 nM) for somatostatin. The biphasic nature of the dissociation process confirmed the heterogeneity of somatostatin binding sites. Apart from somatostatin, no peptide (1 microM) tested influenced the binding of 125I-Tyr11-somatostatin. The present data represent the first analysis of somatostatin binding sites in lung.

Animals↗

Effect of sensitization on somatostatin concentration and binding in cytosol from guinea pig airways.

In guinea pigs sensitized with 1 microgram ovalbumin together with 100 mg Al(OH)3, somatostatin levels were selectively increased up to two and 3 times in tissue extracts from trachea and bronchi, respectively, but not in lung as compared to controls. The increase of somatostatin levels observed in trachea and bronchi after sensitization was associated with a decrease in the binding capacity of both high- and low-affinity binding sites (without changes in the affinity values) in the corresponding cytosolic fractions. These results suggests that an increase in airways somatostatin content may be involved in the pathogenesis of anaphylactic bronchoconstriction.

Anaphylaxis↗

Tissue and plasma distribution of exogenous growth hormone-releasing factor analogue (GRF1-29NH2) after intravenous, subcutaneous and intraperitoneal injection in the rat.

1. The administration of 125I-labelled growth hormone-releasing factor (GRF) analogue 1-29NH2 by intravenous, subcutaneous or intraperitoneal injection to rats leads to rapid (i.v.) or slow (s.c. and i.p.) increases in plasma radioactivity followed by extensive breakdown of the peptide. 2. Tissues possessing GRF-like immunoreactivity such as gastric antrum (but not fundus), duodenum and ileum showed in vitro specific uptake of 125I-GRF probably mediated by vasoactive intestinal peptide (VIP) receptors. 3. Pituitary (the primary target organ for GRF) but neither thyroid nor parathyroid exhibited specific uptake of 125I-GRF.

Animals↗

Effect of gastroduodenostomy on intestinal vasoactive intestinal peptide (VIP) levels, and VIP binding and VIP stimulation of cyclic AMP in intestinal epithelial cells from rat.

The concentration of VIP in duodenum and jejunum as well as the interaction of VIP (binding and stimulation of cyclic AMP accumulation) with epithelial cells from both gut segments were studied in rats after surgical bypass of the pylorus by gastroduodenostomy. Duodenal VIP concentration increased in rats with gastroduodenostomy as compared to sham-operated animals. The binding capacity (but not the affinity) of VIP binding sites and the efficiency (but not the potency) of VIP on cyclic AMP accumulation decreased in the condition of gastroduodenostomy. However, no modifications in either VIP concentration and interaction could be seen at the jejunal level.

1-Methyl-3-isobutylxanthine↗

Ileal vasoactive intestinal peptide (VIP) levels and VIP receptor/effector system in ileal epithelial cells after colectomy in the rat.

The interaction of VIP (binding to specific receptors and stimulation of cyclic AMP accumulation) with ileal epithelial cells and the levels of the neuropeptide in the ileal segment were determined after colectomy (removal of cecum and colon followed by ileorectal anastomosis) in the rat. The number of VIP receptors (but not the affinity) and the efficiency (but not the potency) of the neuropeptide upon stimulation of cyclic AMP accumulation in ileal epithelial cells increased 21 (but not 7) days after colectomy, whereas VIP ileal levels followed an inverse pattern. These changes could be interpreted in terms of a consequence or a cause of some of the phenomena that appear after colectomy, i.e., chronic watery diarrhea.

Animals↗

Identification and characterization of specific binding sites for somatostatin in cytosol of bovine cystic duct mucosa.

Specific binding sites for somatostatin have been detected in cytosolic fraction of bovine cystic duct mucosa. At 37 degrees C, the interaction of 125I-Tyr11-somatostatin with cytosolic fraction was rapid, reversible, specific and saturable. At equilibrium, the binding of tracer was competitively inhibited by native peptide in the 1 nM to 2 microM range of concentrations. Scatchard analysis of binding data suggested the presence of two distinct classes of somatostatin binding sites: a class with a high affinity (Kd = 7.8 +/- 0.3 nM) and a low capacity (1.3 +/- 0.3 pmol somatostatin/mg protein) and a class with a low affinity (Kd = 129.1 +/- 2.0 nM) and a high capacity (43.5 +/- 6.7 pmol somatostatin/mg protein). The binding sites were shown to be highly specific for somatostatin since neuropeptides present in cystic duct such as Leu-enkephalin, neurotensin, substance P and vasoactive intestinal peptide did practically not show competition. These findings suggest that somatostatin could contribute to the regulation of the functions of the cystic duct mucosa in physiological and pathological conditions.

Animals↗

Identification of insulin receptors in epithelial cells from duodenum, jejunum, ileum, caecum, colon and rectum in the rat.

Specific insulin binding was demonstrated in isolated epithelial cells of rat intestine, from the duodenum to the rectum. In every segment tested, insulin binding exhibited similar properties relating to kinetics and specificity and could be interpreted in terms of two populations of receptors possessing different affinities and binding capacities. Insulin receptors were unequally distributed along the intestinal tract, suggesting that the hormone may have varying importance at different points along the intestinal axis. These observations together with previous data on insulin regulation of a number of intestinal epithelial activities suggest that the pancreatic hormone may play an important role in gut physiology from the duodenum to the rectum.

Animals↗

Effect of small intestinal resection on somatostatin binding to cytosol of rabbit gastric mucosa.

Small bowel resection in the rabbit increased gastric (fundus and antrum) somatostatin content and decreased the number of somatostatin binding sites (but not their corresponding affinity values) in gastric (fundus and antrum) cytosol three weeks after surgery. Five weeks after resection the number of somatostatin binding sites at both fundic and antral levels as well as antral somatostatin content returned towards control values whereas the fundic concentration of the peptide remained increased. Present results together with the known inhibitory role of somatostatin on various gastric functions suggest that the gastric alterations showed by animals subjected to small bowel resection may be due, at least in part, to the observed decrease of the number of gastric somatostatin binding sites.

Animals↗

Effects of age and androgens upon functional vasoactive intestinal peptide receptors in rat prostatic epithelial cells.

The specific binding of vasoactive intestinal peptide (VIP) and the stimulatory effect of VIP upon cyclic AMP accumulation in isolated epithelial cells of rat ventral prostate were age dependent. The number of VIP receptors decreased but the efficiency of VIP on cyclic AMP accumulation increased in prostatic epithelium when considering the periods 35-65 days and 3-6 months. Since these features could be related to the known age-related decrease of androgen and androgen-receptor levels, we studied the effect of testosterone and its 5 alpha-reduced metabolite dihydrotestosterone upon both steps of VIP action. The two steroid hormones exerted a non-competitive inhibition on VIP-induced cyclic AMP accumulation but did not modify VIP binding to its specific receptors. This modulatory effect of androgens might involve their interaction with specific sites on the cell membrane leading to modifications of membrane activities including adenylate cyclase, as has been suggested by an increasing number of recent reports.

Aging↗

Characterization of somatostatin binding sites in cytosolic fraction of pig retina.

Specific binding sites for somatostatin have been characterized in the cytosolic fraction of the pig retina. The interaction of 125I-Tyr11-somatostatin with cytosolic fraction was rapid, reversible, specific, saturable and temperature-dependent. At 15 degrees C, the binding data were compatible with the existence of two classes of binding sites: a high-affinity class with a Kd of 16.1 nM and a low-affinity class with a Kd of 236.4 nM. The presence of somatostatin binding sites in the cytosolic fraction of the retina, together with the known occurrence of somatostatin at this level, strongly suggest that this peptide may be involved in the physiology of the retina.

Animals↗