Increase In Basal Secretion Research Articles

Endocrine patterns of the post-partum cow

Summary. Milked dairy cows generally have a shorter post-partum interval to ovarian cyclicity than suckling dairy or beef cows. In milked and suckling cows, there is a strong seasonal influence with spring-calving cows remaining anoestrous longer. Increasing the suckling intensity further delays the onset of ovarian cyclicity, probably by increasing the frequency or strength of its inhibitory influence on hypothalamic activity. Plasma FSH levels rise in most cows 5–10 days after calving and thereafter the random changes observed have little relationship to the onset of cycles. Recovery of FSH release therefore occurs earlier post partum than recovery of LH release. Hyperprolactinaemia is not a cause of reproductive failure in milked or suckling cows because there is no correlation between plasma prolactin levels and the onset of ovarian cycles. Plasma LH concentrations undergo significant changes directly related to the initiation of ovarian cycles, with low plasma levels immediately post partum, followed by an increase in basal secretion and the development of clear LH episodes. This pulsatile pattern appears earlier in dairy than in beef cows and is further delayed by suckling compared to milking. Before the first ovulation there is an increased frequency and peak height of LH episodes leading to a rise in plasma LH levels and eventually to a preovulatory-type LH surge which results in the first ovulation. These changes in the pattern of LH release appear definitive in the initiation of ovarian activity in post-partum cows.

Lipid metabolism and production of incretins in patients with obesity and type 2 diabetes mellitus after biliopancreatic diversion in Hess-Marceau modification at the early stages of follow-up

Objectives: To study the lipid metabolism and secretion of GLP-1 and GIP in obese patients with type 2 diabetes before and 3 months after biliopancreatic diversion (BPD) in Hess-Marceau modification.Materials and methods: 22 patients (44 [38.0; 55.0] years) with morbid obesity (BMI of 50.1 [41.3; 53.8]) and type 2 diabetes were included in the study. Total cholesterol, LDL-C, HDL-C, TG, GLP-1, GIP were evaluated before and 3 months after BPD.Results: At 3 months after BPD there were decreases in weight by 15.8% and a reduction in total cholesterol levels and LDL-C, an increase in the number of patients with a target value of total cholesterol and LDL-C (from 4/22 (18%) to 20/22 (91%, p = 0.0001)). TG levels during the first 3 months after BPD did not change, but there was a trend to an increase in the number of patients with TG target levels (from 8/22 (32%) to 15/22 (68%, p = 0.058)). Due to the reduction of total cholesterol intake after malabsorbtive surgery, we saw a regular decrease in HDL-cholesterol. 3 months after BPD there was an increase in basal secretion of GLP-1 and GIP as compared to preoperative levels, and a strong positive association of the basal GIP with the HDL-C (r = 0,8, p = 0.007) and a strong negative association – with the atherogenic index (r = – 0,7, p = 0.009) at 3 weeks after surgery.Conclusions: The majority of patients with obesity and type 2 diabetes as early as at 3 months after BPD in the modification by Hess-Marceau achieve a significant improvement in the parameters of lipid metabolism; expect positive impact of the GIP on lipid metabolism.

CFTR Knockdown induces proinflammatory changes in intestinal epithelial cells.

BackgroundHyperinflammation is a hallmark feature of cystic fibrosis (CF) airways. However, inflammation has also been documented systemically and, more recently, in extrapulmonary CF-affected tissues such as the pancreas and intestine. The pathogenesis of CF-related inflammation and more specifically the role of the cystic fibrosis transmembrane conductance regulator (CFTR) in that respect are not entirely understood. We have tested the hypothesis that genetic depletion of CFTR will affect the inflammatory status of human intestinal epithelial cell lines.MethodsCFTR expression was genetically depleted from Caco-2/15 and HT-29 cells using short hairpin RNA interference (shRNAi). Inflammatory conditions were induced by the addition of human recombinant tumor necrosis factor (TNF) or Interleukin-1β (IL-1β) for various periods of time. Gene expression, mRNA stability and secreted levels of interleukin (IL)-6, −8 and 10 were assessed. Analysis of pro- and anti-inflammatory signaling pathways including mitogen-activated protein kinases (p38, ERK 1/2 and JNK), nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor alpha (IκBα), and nuclear factor-kappa B (NF-κB) was also performed. Eosinophils were counted in the jejunal mucosa of Cftr−/− and Cftr+/+ mice.ResultsCFTR gene and protein knockdown caused a significant increase in basal secretion of IL-8 as well as in IL-1β-induced secretion of IL-6 and −8. Release of the anti-inflammatory cytokine, IL-10, remained unaffected by CFTR depletion. The enhanced secretion of IL-8 stems in part from increased IL8 mRNA levels and greater activation of ERK1/2 MAPK, IκBα and NF-κB in the CFTR knockdown cells. By contrast, phosphorylation levels of p38 and JNK MAPK did not differ between control and knockdown cells. We also found a higher number of infiltrating eosinophils in the jejunal mucosa of Cftr −/− females, but not males, compared to Cftr +/+ mice, thus providing in vivo support to our in vitro findings.ConclusionCollectively, these data underscore the role played by CFTR in regulating the intestinal inflammatory responses. Such findings lend support to the theory that CFTR exerts functions that may go beyond its role as a chloride channel whereby its disruption may prevent cells to optimally respond to exogenous or endogenous challenges. These observations are of particular interest to CF patients who were found to display alterations in their intestinal microbiota, thus predisposing them to pathogens that may elicit exaggerated inflammatory responses.Electronic supplementary materialThe online version of this article (doi:10.1186/s12950-015-0107-y) contains supplementary material, which is available to authorized users.

Novel features on the regulation by mitochondria of calcium and secretion transients in chromaffin cells challenged with acetylcholine at 37°C.

From experiments performed at room temperature, we know that the buffering of Ca2+ by mitochondria contributes to the shaping of the bulk cytosolic calcium transient ([Ca2+]c) and secretion transients of chromaffin cells stimulated with depolarizing pulses. We also know that the mitochondrial Ca2+ transporters and the release of catecholamine are faster at 37°C with respect to room temperature. Therefore, we planned this investigation to gain further insight into the contribution of mitochondrial Ca2+ buffering to the shaping of [Ca2+]c and catecholamine release transients, using some novel experimental conditions that have not been yet explored namely: (1) perifusion of bovine chromaffin cells (BCCs) with saline at 37°C and their repeated challenging with the physiological neurotransmitter acetylcholine (ACh); (2) separate blockade of mitochondrial Ca2+ uniporter (mCUP) with Ru360 or the mitochondrial Na+/Ca2+ exchanger (mNCX) with CGP37157; (3) full blockade of the mitochondrial Ca2+ cycling (mCC) by the simultaneous inhibition of the mCUP and the mNCX. Ru360 caused a pronounced delay of [Ca2+]c clearance and augmented secretion. In contrast, CGP37157 only caused a tiny delay of [Ca2+]c clearance and a mild decrease in secretion. The mCC resulting in continued Ca2+ uptake and its release back into the cytosol was interrupted by combined Ru360 + CGP37157 (Ru/CGP), the protonophore carbonyl cyanide‐p‐trifluoromethoxyphenylhydrazone, or combined oligomycin + rotenone (O/R); these three treatments caused a mild but sustained elevation of basal [Ca2+]c that, however, was not accompanied by a parallel increase in basal secretion. Nevertheless, all treatments caused a pronounced augmentation of ACh‐induced secretion, with minor changes of the ACh‐induced [Ca2+]c transients. Combined Ru/CGP did not alter the resting membrane potential in current‐clamped cells. Additionally, Ru/CGP did not increase basal [Ca2+]c near subplasmalemmal sites and caused a mild decrease in the size of the readily releasable vesicle pool. Our results provide new functional features in support of the view that in BCCs there are two subpopulations of mitochondria, M1 underneath the plasmalemma nearby exocytotic sites and M2 at the core cell nearby vesicle transport sites. While M1 serves to shape the ACh‐elicited exocytotic response through its efficient Ca2+ removal by the mCUP, M2 shapes the lower [Ca2+]c elevations required for new vesicle supply to the exocytotic machinery, from the large reserve vesicle pool at the cell core. The mCUP of the M1 pool seems to play a more prominent role in controlling the ACh responses, in comparison with the mNCX.

Progastrin Is Directed to the Regulated Secretory Pathway by Synergistically Acting Basic and Acidic Motifs

Bioactivation of prohormones occurs in the granules of the regulated secretory pathway of endocrine cells, which release hormones in response to external stimulation. How secretory granules are formed and how the cargo is selected is still unclear, but it has been shown for several prohormones and processing enzymes that domains within the prohormone structure can act as "sorting signals" for this pathway. The domains mediate interactions with other proteins or with the membrane or facilitate aggregation of the (pro)peptides. We have now searched for domains in progastrin that are active in sorting the prohormone into secretory granules. Truncation studies showed that the N-terminal 30 residues of progastrin are dispensable, whereas the last 49 residues are sufficient for correct biosynthesis of bioactive gastrin. Thus, further N-terminal truncation abolished gastrin expression. C-terminal truncation of 8 residues resulted in an increase in basal secretion as did point mutations in the dibasic processing sites of progastrin. These mutants, however, still responded to secretagogues, suggesting a residual sorting capacity to the regulated pathway. Amino acid substitutions in an acidic, polyglutamate motif within gastrin-17, the main bioactive, cellular gastrin form, did not alter secretion per se, but when these residues were substituted in C-terminally truncated mutants, double mutants increased in basal secretion and did not respond to secretagogue stimulation. This implies that the mutants are constitutively secreted. Our data suggest that the dibasic processing sites constitute the most important sorting domain of progastrin, and these sites act in synergy with the acidic domain.

Secretory activity and cell cycle alteration of alveolar type II cells in the early and late phase after irradiation

Purpose: Type II cells and the surfactant system have been proposed to play a central role in pathogenesis of radiation pneumonitis. We analyzed the secretory function and proliferation parameters of alveolar type II cells in the early (until 24 h) and late phase (1–5 weeks) after irradiation (RT) in vitro and in vivo. Methods and Materials: Type II cells were isolated from rats according to the method of Dobbs. Stimulation of secretion was induced with terbutaline, adenosine triphosphate (ATP), and 12-O-tetradecanoylphorbol-13-acetate (TPA) for a 2-h period. Determination of secretion was performed using 3H-labeled phosphatidylcholine. For the early-phase analysis, freshly isolated and adherent type II cells were irradiated in vitro with 9–21 Gy (stepwise increase of 3 Gy). Secretion stimulation was initiated 1, 6, 24, and 48 h after RT. For late-phase analysis, type II cells were isolated 1–5 weeks after 18 Gy whole lung or sham RT. Each experiment was repeated at least fivefold. Flow cytometry was used to determine cell cycle distribution and proliferating cell nuclear antigen index. Results: During the early-phase (in vitro) analysis, we found a normal stimulation of surfactant secretion in irradiated, as well as unirradiated, cells. No change in basal secretion and no dose effect were seen. During the late phase, 1–5 weeks after whole lung RT, we observed enhanced secretory activity for all secretagogues and a small increase in basal secretion in Weeks 3 and 4 (pneumonitis phase) compared with controls. The total number of isolated type II cells, as well as the rate of viable cells, decreased after the second post-RT week. Cell cycle alterations suggesting an irreversible G 2/M block occurred in the second post-RT week and did not resolve during the observation period. The proliferating cell nuclear antigen index of type II cells from irradiated rats did not differ from that of controls. Conclusion: In contrast to literature data, we observed no direct effect of radiation on secretory activity in the early phase after RT. In our study of isolated type II cells, as well as in intact animals, RT did not result in an impaired surfactant secretion up to 5 weeks after RT. Our in vivo experiments even showed an increased response of phosphatidylcholine secretion to all known secretagogues at Weeks 3 and 4 after whole lung RT, possibly due to inflammatory cytokines. Cell cycle alterations with G 2/M block and cell loss in the late post-RT period may contribute more to the manifestation of radiation-induced lung damage than functional impairment in type II cells.

Growth Hormone Secretagogue (GHS) Analogue, Hexarelin Stimulates GH From Peripheral Lymphocytes

The role of growth hormone releasing hormone (GHRH) and growth hormone releasing peptide-6 (GHRP-6) analogue hexarelin was investigated in the regulation of GH production from lymphocytes. Porcine and bovine blood mononuclear cells were separated using density gradient centrifugation method by layering the whole blood or buffy coat cells on lymphodex. Cells were incubated for 3 or 5 days with or without phytohemagglutinin (PHA-M), GHRH, GHRP-6 analogue hexarelin, somatostatin or GHRH + hexarelin. Growth hormone was fractionated from supernatants by gel chromatography and further concentrated by lyophilization at - 20 degrees C. A nearly two fold increase in basal secretion of GH (porcine: 3.5 +/- 0.1 ng/ml, bovine: 3.2 +/- 0.2 ng/ml) was achieved by GHRH and hexarelin at concentrations of 0.1, 1.0, 10 and 100 nM in both porcine and bovine cells. Lymphocytic GH release was also stimulated in response to PHA-M (10 micro g/well). Neither a dose dependent nor a synergistic nor an additive effect was apparent on GH secretion from lymphocytes. GHRH stimulated lymphocytic GH secretion, whereas, somatostatin had no effect. This study reports for the first time that hexarelin stimulates the secretion of GH from peripheral lymphocytes.

Osmotic tolerance limits of canine pancreatic islets.

Future improvements in the recovery and function of pancreatic islets following cryopreservation will require a more precise quantification of the stresses that occur at each stage of the cryopreservation protocol. Changes in solution osmolality during the addition and dilution of cryoprotectants and during freezing and thawing induce changes in islet volume that may exceed tolerable limits. The aim of this study was to determine the range of solution osmolalities that results in significant changes in islet function. Islets were isolated from canine pancreases by collagenase digestion and Euro-Ficoll purification. Following 12-h culture at 37 degrees C, islets were counted and dispensed into multiwell plate inserts. Islet function was assessed in each well immediately before and 24 h following a 10-min osmotic challenge with hypo- or hyperosmotic solutions of PBS (0, 75, 150, 300, 600, 1200, or 2300 mOsm/kg) at 22 degrees C. Canine islets reached their osmotic equilibrium within 10 min. Duplicate wells were used for each osmolality treatment for each of six donors (n = 12). No significant differences in basal or glucose-stimulated insulin secretion were found between wells prior to the osmotic challenge (3.35 +/- 0.45 and 20.98 +/- 3.36 microIU/IE/h, respectively). Following the osmotic challenge and 24-h in vitro tissue culture, a significant increase in basal secretion was observed for islets exposed to 0 and 75 mOsm/kg solutions and a significant decrease for islets exposed to 2300 mOsm/kg solution. Islets exposed to 0 and 2300 mOsm/kg solutions showed significant decreases in the stimulated insulin secretion when compared to controls. Solution osmolalities of 150-1200 mOsm/kg appear to be tolerated by canine islets with no significant deviations in insulin secretion. The corresponding tolerable volume range was 152.6 +/- 6.8% to 60 +/- 5.1% of the isotonic islet volume. The minimum critical volume was used in a theoretical analysis of the islet volumes that would result from equilibrium freezing in dimethyl sulfoxide (DMSO). The calculations show that 1.5 mol/l DMSO is sufficient to prevent damage to islets due to excessive shrinkage. Further refinement of cryoprotectant addition and dilution protocols, and cooling and warming protocols for canine islets, are now possible.

Testosterone deficiency in young men: marked alterations in whole body protein kinetics, strength, and adiposity.

To investigate specific effects of androgens on whole body metabolism, we studied six healthy lean men (mean +/- SEM age, 23.2 +/- 0.5 yr) before and after gonadal steroid suppression with a GnRH analog (Lupron), given twice, 3 weeks apart. Primed infusions of [13C]leucine, indirect calorimetry, isokinetic dynamometry, growth factor measurements, and percutaneous muscle biopsies were performed at baseline (D1) and after 10 weeks of treatment (D2); each subject served as his own control. Testosterone concentrations were markedly suppressed after 10 weeks of treatment (D1, 535 +/- 141 ng/dL; D2, 31 +/- 9). Leucine's rate of appearance (index of proteolysis) was markedly suppressed after 10 weeks of hypogonadism (-13%; P = 0.01) as well as the nonoxidative leucine disposal, an index of whole body protein synthesis (-13%; P = 0.01) without any changes in plasma amino acid concentrations. All subjects studied after 10 weeks showed a decrease in fat-free mass, as measured by skinfold calipers and dual emission x-ray absortiometry scans (D1, 56.5 +/- 2.9 kg; D2, 54.4 +/- 2.5; P = 0.005), and an increase in percent fat mass (D1, 19.2 +/- 2.5%; D2, 22.2 +/- 2.5; P = 0.001). Rates of lipid oxidation decreased (-31%; P = 0.05) after treatment, with parallel changes in resting energy expenditure (-9%; P = 0.05). Mean and peak GH concentrations (measured every 10 min for 6 h) and GH production rates did not decrease after testosterone deficiency, with an actual increase in basal secretion (P < 0.02). Plasma insulin-like growth factor I (IGF-I) concentrations did not change significantly after 10 weeks of treatment (D1, 227 +/- 44 micrograms/L; D2, 291 +/- 60; P = 0.08). Isokinetic dynamometry of leg extensors at 60 degrees and 180 degrees/s was also decreased after 10 weeks of hypogonadism. Total ribonucleic acid (RNA) was isolated from muscle biopsy samples, and ribonuclease protection assays were performed using human complementary DNA clones for IGF-I, IGF-binding protein-4, myosin, and actin. Ten weeks after Lupron treatment, messenger RNA (mRNA) concentrations of IGF-I decreased significantly, whereas there was a trend toward higher IGF-binding protein-4 concentrations, with no change in myosin or actin mRNA concentrations. In conclusion, testosterone deficiency in young men is associated with a marked decrease in measures of whole body protein anabolism, decreased strength, decreased fat oxidation, and increased adiposity. These effects of testosterone deficiency are independent of changes in peripheral GH production and IGF-I concentrations, even though im IGF-I mRNA concentrations decrease. These data suggest a direct effect of androgens on whole body lipid and protein metabolism.

Changes in β amyloid precursor protein secretion associated with the proliferative status of CNS derived progenitor cells

We characterized the secretion of amyloid precursor protein (APP) in rat primary neurons and conditionally immortalized central nervous system (CNS) derived progenitor cell lines, to evaluate their suitability as models for studies on APP metabolism. We observed that primary cells dissociated from different regions of the embryonic brain secreted progressively more APP during in vitro maturation. The increase in basal secretion resulted in an offset of the response to protein kinase C (PKC) activated secretion and was correlated with an increased PKCα expression. The same type of analysis was performed on CNS derived conditionally immortalized cells whose growth potential becomes restricted upon shifting of the culture conditions to a non-permissive temperature (39°C), an event that coincides with their progression toward differentiation. With a pattern consistent with that observed in primary neurons, conditionally immortalized cells exposed to 39°C showed an offset of the secretory response to activation by PdBu and also an increased expression of PKCα. These data suggest that APP secretion and its regulation in CNS derived cells is influenced by the proliferative status of the cells.

Pituitary adenylate cyclase-activating polypeptide stimulates cardiodilatin/atrial natriuretic peptide (CDD/ANP-(99–126) secretion from cultured neonatal rat myocardiocytes

It has been reported that the highly homologous neuropeptides pituitary adenylate cyclase-activating peptide (PACAP) and vasoactive intestinal polypeptide (VIP) exert similar cardiovascular effects in vivo. In the present study we compared the effects of these neuropeptides on myocardial cyclic AMP content and the release of immunoreactive CDD/ANP-(99–126) (atrial natriuretic peptide). In cultured neonatal rat cardiomyocytes PACAP and VIP evoke concentration-dependent increases in intracellular cyclic AMP content but responses to VIP are markedly less. PACAP stimulates the release of CDD/ANP-(99–126) in a concentration-dependent manner with a threshold concentration of 1 nM, and up to a 6-fold increase in basal secretion at 1 μM PACAP. In contrast, VIP had no effect on the release of CDD/ANP. Pretreatment of cells with the competitive PACAP-antagonist, PACAP-6-38 (1 μM), significantly reduces the effects of PACAP on intracellular cyclic AMP and on CDD/ANP-(99–126) secretion and abolishes the effects of VIP on cyclic AMP. Pretreatment with VIP-receptor antagonist (1 μM) prevents the cyclic AMP-responses to VIP while increases in cyclic AMP as well as stimulation of CDD/ANP-(99–126) release by PACAP are not affected. It is concluded that both neuropeptides directly influence cardiac myocytes through an increase in intracellular cyclic AMP. Release of CDD/ANP-(99–126) by PACAP may be involved in the decrease in blood pressure that follows intravenous administration of this peptide. The higher potency of PACAP to induce cyclic AMP synthesis, its stimulating effect on the release of CDD/ANP-(99–126) and the finding that the VIP-receptor antagonist inhibits responses to VIP but not to PACAP suggest that PACAP activates cardiac myocytes through a PACAP-specific receptor.

Pertussis toxin pretreatment enhances catecholamine secretion induced by pituitary adenylate cyclase-activating polypeptide in cultured porcine adrenal medullary chromaffin cells: A possible role of the inositol lipid cascade

We determined how pertussis toxin (PTX) pretreatment alters PACAP-induced catecholamine secretion in cultured porcine adrenal medullary cells. Pretreatment of these cells with PTX (1 ng/ml for 24 h or 10 ng/ml for 6 h) markedly enhanced PACAP-induced catecholamine secretion. PTX pretreatment also produced a small increase in basal secretion and secretion in response to nicotine and carbachol, but the effect of the PACAP-induced secretion was most striking. We examined the role of the phosphoinositol cascade in potentiating the PACAP-induced catecholamine secretion by PTX and found that PACAP-induced accumulation of inositol phosphates in PTX-pretreated cells was significantly greater than that in untreated cells. Furthermore, removal of extracellular Ca 2+ and addition of Ca 2+ channel blockers inhibited the catecholamine secretion induced by PACAP in PTX-pretreated cells. From these results, we speculate that a PTX-sensitive G-protein tonically inhibits phospholipase C. PTX enhances the PACAP-induced secretion of catecholamine by blocking the action of this inhibitory G-protein.

Agonist-Induced Release of Gonadotropin-Releasing Hormone, Luteinizing Hormone, and Follicle-Stimulating Hormone and their Associations with Basal Secretion of Luteinizing Hormone and Follicle-Stimulating Hormone throughout Lactation in Sows

Our working hypothesis was that the normal increase in basal secretion of LH and FSH during lactation in sows is paralleled by an increase in readily releasable pools of GnRH, LH, and FSH. Sows fitted with indwelling catheters and nursing 9-11 piglets were sampled on Days 1, 7, 14, and 21 (primiparous sows; n = 10; experiment 1) or Days 7 and 21 (multiparous sows; n = 10; experiment 2) of lactation. On each day, blood was collected at 10-min intervals for 2 h before one member of a pair of sows was infused (i.v.) with saline (n = 5) and the other with the neuroexcitatory amino acid N-methyl-DL-aspartic acid (NMA; 10 mg/kg b.w.) to measure readily releasable GnRH as estimated by LH release. Two hours later, each sow that had received saline was given GnRH (100 micrograms, i.v.) for estimation of releasable pools of pituitary LH and FSH. Responses to NMA and GnRH were estimated by area under the curve (ng.ml-1.min) of serum LH profiles for 1 h (NMA group) or 2 h (GnRH group) after treatment. Basal LH secretion in primiparous sows decreased from Day 1 to Day 7 and then returned to Day 1 values by Day 21. Similarly, in multiparous sows, basal LH was lower on Day 7 than on Day 21. Basal FSH secretion increased linearly as lactation progressed in primiparous and multiparous sows.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of ozone on the cholinergic secretory responsiveness of ferret tracheal glands

Oxidant air pollutants exacerbate several pulmonary diseases. Inhalation of ozone has been shown to induce airway smooth muscle hyperresponsiveness. Oxidant injury could also affect airway secretory mechanisms. We postulated that oxidant exposure would alter the glycoconjugate secretory function of airway submucosal glands. To test this hypothesis we examined the effects of in vivo ozone exposure on the in vitro secretory responsiveness of ferret tracheal glands. Ferrets were exposed to 1 ppm ozone, 24 hr/day for 3 or 7 days. Following exposure, glandular explants, denuded of surface epithelial cells, were prepared and incubated in medium containing 3H-glucosamine for 18 hr. Basal secretion of labeled glycoconjugates was significantly increased 31% following 3 days of ozone exposure (P less than or equal to 0.05) and remained elevated 11% after 7 days of exposure compared to the air-exposed group. After 3 or 7 days of exposure to ozone, tracheal gland responsiveness to carbachol was increased as indicated by significantly lower EC50 values (log molar concentration) of -6.43 +/- 0.04 (n = 6) and -6.50 +/- 0.11 (n = 5), respectively; compared to -6.20 +/- 0.08 (n = 6) for the air-exposed group. There was no difference in carbachol EC50 values for air and 7-day ozone-exposed animals treated with dexamethasone. Dexamethasone did not attenuate the ozone-induced increase in basal secretion. Tracheal gland responsiveness to alpha- or beta-adrenergic agonists was not changed by oxidant exposure. These experiments suggest that oxidant injury not only increases basal secretion of respiratory glycoconjugates but also increases tracheal gland sensitivity to a cholinergic agonist.

Extracellular sodium is required for methacholine‐induced secretion of mucus glycoconjugates from canine tracheal explants

Extracellular sodium is known to influence secretion by certain secretory cells, possibly by mobilizing calcium from cellular stores or by altering intracellular pH via regulation of a Na(+)-H+ antiport system. Using canine tracheal explants, we determined whether agents which alter sodium fluxes are capable of modulating basal or cholinergically-induced secretion of mucus glycoconjugates. Methacholine, a cholinergic agonist, increased mucus secretion from explants incubated in the presence or absence of calcium, but had no effect on secretion when incubated in sodium-deficient media, indicating (a) that cholinergically-induced secretion can be mediated by mobilization of cellular calcium and (b) that extracellular sodium was required for this stimulatory effect. Several agents which increase intracellular sodium were tested for their effect on mucus secretion. Ouabain, a sodium pump inhibitor, and veratridine, a sodium channel activator, did not significantly affect control or methacholine-induced secretion; gramicidin, a sodium ionophore, also had no effect on basal release. Tetrodotoxin, a sodium channel inhibitor, was also without effect on basal or methacholine-stimulated mucus release. Agents which alter intracellular pH were also examined for their effects on basal or methacholine-induced glycoconjugate secretion. Amiloride, which decreases intracellular pH by inhibiting Na(+)-H+ exchange, produced a 19 per cent increase in basal secretion (not statistically significant), but had no effect on methacholine-induced secretion. An agent, 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS), which decreases intracellular pH by inhibiting HCO3(-)-Cl- exchange, elicited decreases in both basal and methacholine-induced secretion, but the inhibition did not reach statistical significance.(ABSTRACT TRUNCATED AT 250 WORDS)

The diacylglycerol kinase inhibitor, R59022, potentiates cholecystokinin‐induced enzyme secretion from rabbit pancreatic acini

The putative inhibitor of diacylglycerol kinase activity, 6-(2-[(4-fluorophenyl)phenylmethylene]-1-piperidinyl)-ethyl-7-meth yl-5H- thiazolo[3,2-a]pyrimidin-5-one (R59022), markedly potentiated cholecystokinin-C-terminal-octapeptide(CCK-8-)stimulated enzyme secretion from isolated rabbit pancreatic acini. Maximal potentiation occurred when acini were stimulated in the presence of 5-10 microM R59022. Potentiation depended both on the concentration of R59022 and CCK-8. No potentiation was observed when acini were half-maximally stimulated, whereas the secretory response to maximal and supramaximal concentrations of secretagogue was increased by 50-60%. R59022 alone had no effect on basal enzyme secretion and the drug did not potentiate the secretory response to the Ca2+ ionophore A23187 or to the phorbol ester, 12-O-tetradecanoylphorbol 13-acetate. Moreover, no increase in basal secretion was observed when acini were incubated in the presence of both R59022 and forskolin. These observations strongly suggest that receptor-mediated activation of the inositol phospholipid pathway is required for R59022-induced potentiation. R59022 inhibited the CCK-8-stimulated incorporation of 32Pi into phosphatidic acid dose dependently, without affecting the CCK-8-stimulated hydrolysis of 32P-labelled phosphatidylinositol 4,5-bisphosphate. This is consistent with an inhibitory effect of R59022 on acinar cell diacylglycerol kinase activity. The potentiating effect of R59022 was mimicked by 12-O-tetradecanoylphorbol 13-acetate added simultaneously with CCK-8. Therefore, it is concluded that in the presence of 5-10 microM R59022 the receptor-mediated increase in acinar cell diacylglycerol content is enhanced leading to enhanced activation of protein kinase C and to potentiation of the secretory response. The fact that the secretory response to maximal and supramaximal concentrations of CCK-8 is potentiated by R59022 suggests that at these concentrations of secretagogue the diacylglycerol/protein kinase C branch of the signal-transduction route is rate-limiting.

Effect of acute ethanol on beta-endorphin secretion from rat fetal hypothalamic neurons in primary cultures

To characterize the effect of ethanol on the hypothalamic β-endorphin-containing neurons, rat fetal hypothalamic neurons were maintained in primary culture, and the secretion of β-endorphin (β-EP) was determined after ethanol challenges. Constant exposure to ethanol at doses of 6–50 mM produced a dose-dependent increase in basal secretion of β-EP from these cultured cells. These doses of ethanol did not produce any significant effect on cell viability, DNA or protein content. The stimulated secretion of β-EP following constant ethanol exposure is short-lasting. However, intermittent ethanol exposures maintained the ethanol stimulatory action on β-EP secretion for a longer time. The magnitude of the β-EP response to 50 mM ethanol is similar to that of the β-EP response to 56 mM of potassium. Ethanol-stimulated β-EP secretion required extracellular calcium and was blocked by a calcium channel blocker; a sodium channel blocker did not affect ethanol-stimulated secretion. These results suggest that the neuron culture system is a useful model for studying the cellular mechanisms involved in the ethanol-regulated hypothalamic opioid secretion.

Isoproterenol-induced desensitization of mucin release in isolated rat submandibular acini

Release of [ 14C]glucosamine-labelled mucins was studied in vitro using well-characterised preparations of rat submandibular acini. Mucin release was stimulated by forskolin, an activator of the catalytic subunit of adenylate cyclase, adn 3-isobutyl-1-methylxanthine (IBMX), a cyclic nucleotide phosphodiesterase inhibitor. Both stimulated in a dose-dependent manner to the same maximum as that seen with isoproterenol. Neither forskolin nor IBMX added in the presence of isoproterenol increased secretion above the maximum in response to isoproterenol alone, suggesting a similar mechanism of action, mediated by cyclic AMP. Prior exposure of acini to isoproterenol (10 μM) for 45 min, followed by washout resulted in (a) persistent increase in basal secretion which was abolished by propranolol and (b) reduced stimulation of mucin secretion in response to either a second isoproterenol challenge, noradrenaline or forskolin. Thus, exposure of rat submandibular acini in vitro desensitizes the cells to subsequent stimulation. Although this mimics the decreased β-adrenergic secretory responses seen in submandibular cells from cystic fibrosis patients, results suggest that the isoproterenol-induced desensitization is at the level of β-receptor and adenylate cyclase, rather than distal to cyclic AMP.

SOMATOMEDIN-C (Sm-C/IGF-1) SYNERGISES WITH FIBROBLAST GROWTH FACTOR (FGF) ON SERTOLI CELLS (SC) MULTIPLICATION AND DIFFERENTIATION

We have characterized Sm-C secretion and regulation by immature porcine SC cultured (ESPE 86). The present study evaluates the autocrine action of Sm-C on SC. The presence of IGF type I receptors on SC was identified by binding (Kd = 10−9 M) and cross-linking. No detectable insulin receptor was found. FGF and Sm-C stimulate SC DNA synthesis with observed ED50 of 2.0 and 7.5 ng/ml respectively. Synergistic effect of FGF+Sm-C was found at saturating concentration of both factors. SC multiplication was increased compared to controls by Sm-C, FGF, Sm-C + FGF by a factor of 1.65, 2.05 and 3.2 respectively. When SC function was studied, FSH receptor number was increased by 1.65, 2.05 and 3.2 respectively. When SC function was studied, FSH receptor number was increased by 1.6 by FGF alone with no effect per se nor synergistic of Sm-C. FSH stimulated cAMP secretion by SC was enhanced by FGF (140%), FGF+Sm-C (195%), but not Sm-C alone. When Plasminogen Activator-secretion by SC was measured, a 3.0 and 22 time increase in basal secretion was observed with FGF and FGF+Insulin respectively compared with controls; furthermore, in the presence of FGF and FGF+Insulin, additional 3.3 and 1.24 respective increment factors were observed after FSH stimulation. All the insulin effects were observed at 5 μg/ml and are likely to be mediated through IGF type 1 receptors. Conclusions: These data demonstrate that Sm-C/IGF-1 secreted by SC may have an autocrine effect expressed by the stimulation of SC multiplication and by the synergistic effect with FGF on SC function. These effects of Sm-C on SC, combined to its paracrine effects on Leydig cells (ESPE 86) emphasize the complex but key actions of Sm-C on testicular function and its maturation.

Effect of Small Bowel Bypass on Serum Gastrin Levels and Gastric Acid Secretion in Man

In animals massive resection of the small intestine is followed by increased gastric acid secretion and an increase in serum gastrin levels. Whether hypersecretion occurs in man after intestinal resections or intestinal bypass is unclear, but an increase in fasting gastrin levels has been reported after intestinal resection. In this series a significant increase in basal gastric acid secretion and fasting serum gastrin levels has been demonstrated after intestinal bypass. However, none of the patients developed peptic ulcer or clinical symptoms of hypersecretion after the bypass operation. Whether the increase in basal secretion and the serum gastrin concentration are interrelated or of any clinical importance is uncertain.