Somatostatin-secreting Cells Research Articles

8704 A DREADD-base Chemogenetic Approach to Quiz the Impact of the Delta Cell Input on Islet Hormone Secretion

Abstract Disclosure: D. Hakim Rodriguez: None. A. Tamayo: None. R. Rodriguez Diaz: None. E. Pereira: None. O. Alcazar: None. M. Makhmutova: None. The utilization of DREADDs (Designer Receptors Exclusively Activated by Designer Drugs) that are gated by clozapine-N-oxide (CNO) enables the selective stimulation of genetically specified cell types in vivo. This chemogenetic technology has been successfully applied in various in vivo studies, including those using mice, to remotely control GPCR signaling in specific cells, such as neurons, glia cells, pancreatic β-cells, or cancer cells. Utilizing DREADDs in the study of pancreatic islets, particularly in the somatostatin-producing δ-cells, could offer insights into the complex role of this hormone. Somatostatin is known to inhibit the secretion of other islet hormones, such as insulin and glucagon, but the detailed mechanisms of its action have not been fully elucidated. To address this, we generated mice enabling the expression of either the activator-DREADD Gq or the inhibitor-DREADD Gi in the mouse somatostatin secreting cells, including the islet δ-cell. Immunohistochemical staining of pancreas sections showed that DREADD were expressed and translocated to the membrane of the islet δ-cells. Perifusion experiments with isolated islets showed that δ-cells responded to CNO stimulation as expected. Islets bearing the DREADD Gq secreted somatostatin in a dose-dependent manner, while somatostatin secretion was inhibited in those expressing DREADD Gi. Secretion of insulin and glucagon were modulated upon activation or inhibition of δ-cells via CNO/DREADD manipulation. CNO-driven pulses of somatostatin stimulation caused asynchronous off responses of neighboring α-and β-cells during the time lapses of somatostatin secretion falloff, resulting in a net increment in the magnitude of both insulin and glucagon secretion. Meanwhile, inhibition of somatostatin secretion surprisingly unleashed glucagon secretion in non-permissive 16mM glucose. These results encouraged us to explore the potential application of DREADD tools in isolated human islets. We will discuss our current approaches whilst using viral vectors to drive the expression of DREADDs in human δ-cells and its impact in the overall islet secretory output. Presentation: 6/2/2024

Computational Reconstruction of Pancreatic Islets as a Tool for Structural and Functional Analysis.

Structural properties of pancreatic islets are key for the functional response of insulin, glucagon, and somatostatin-secreting cells, due to their implications in intraislet communication via electric, paracrine, and autocrine signaling. In this protocol, the three-dimensional architecture of a pancreatic islet is firstly reconstructed from experimental data using a novel computational algorithm. Next, the morphological and connectivity properties of the reconstructed islet, such as the number and percentages of the different type of cells, cellular volume, and cell-to-cell contacts, are obtained. Then, network theory is used to describe the connectivity properties of the islet through network-derived metrics such as average degree, clustering coefficient, density, diameter, and efficiency. Finally, all these properties are functionally evaluated through computational simulations using a model of coupled oscillators. Overall, here we describe a step-by-step workflow, implemented in IsletLab, a multiplatform application developed specifically for the study and simulation of pancreatic islets, to apply a novel computational methodology to characterize and analyze pancreatic islets as a complement to the experimental work.

Analysis of Somatostatin-Secreting Gastric Delta Cells according to Upper Abdominal Symptoms and Helicobacter pylori Infection in Children

PurposeGastric delta cells (D-cells), which are somatostatin-secreting cells, are the main paracrine inhibitor of acid secretion. The number of D-cells was studied in children presenting with upper gastrointestinal (UGI) disease.MethodsWe retrospectively investigated the number of D-cells in the gastric body and antrum through immunofluorescence examinations according to symptoms, endoscopic findings, and Helicobacter pylori infection in 75 children who visited Hanyang University Hospital Pediatrics.ResultsThe mean patient age was 12.2±3.3 years. The male-to-female ratio was 1:1.4. The mean D-cell number per high-power field in the antrum and body was 20.5 and 12 in children with substernal pain, 18.3 and 10.3 in vomiting, 22.3 and 6 in diarrhea, and 9.3 and 6 in abdominal pain, respectively (p>0.05). According to endoscopic findings, the mean D-cell number in the antrum and body was 14.3 and 6 with gastritis, 14 and 9.3 with reflux esophagitis, 16.7 and 8.7 with duodeno-gastric reflux, 19.3 and 12.7 with gastric ulcer, 16 and 13.7 with duodenitis, and 12.3 and 4 with duodenal ulcer, respectively (p>0.05). The D-cell number in the gastric body was 2.7 and 8.7 in children with current H. pylori infection and non-infected children, respectively (p=0.01), while those in the antrum were 15.5 and 14, respectively, with no statistical significance.ConclusionThe D-cell number was lower in the gastric body of children with current H. pylori infection. Further studies concerning peptide-secreting cells with a control group would provide information about the pathogenic pathways of UGI disorder.

A Strong Stomach for Somatostatin.

Evolution of life from single cell to complex multicellular organisms necessitated sophistication in communication between a myriad of specialized cell types. This communication is enabled by a network of neurocrine, endocrine, paracrine, and juxtracrine (contact-dependent) arrangements. The first 3 forms involve release of chemical messengers that activate receptors expressed on the surface or within the target cell to affect a response. Such an array of communication modes enables rapid (eg, neurocrine) and selective (eg, paracrine) communication, while also ensuring a broader reach of chemical substances to all cells within the system (eg, endocrine). One unique chemical within the organism that functions and is regulated by all of the aforementioned modes is somatostatin. Originally isolated from hypothalamic extracts as an inhibitory signal to GH release from the pituitary gland (1, 2), somatostatin was later found to be synthesized by an extensive network of distinct endocrine cells in the gastrointestinal (GI) tract and pancreas (3). Somatostatin subsequently was discovered to function broadly throughout the body as an inhibitor of multiple secretory processes, including insulin and glucagon from the pancreas, gastrin, secretin, cholecystokinin (CCK), vasoactive intestinal peptide, glucagon-like peptide 1, gastric inhibitory peptide, ghrelin, pepsin, and gastric acid from the digestive tract, T3, T4, and calcitonin from the thyroid gland, aldosterone from the adrenal gland, and monoamines from the brain (4, 5). Clinical therapeutics has taken advantage of some of these functions, resulting in the development of various somatostatin analogues for diagnosis, staging and treatment of certain neuroendocrine tumors and for treatment of acromegaly; they also hold promise to treat inflammatory conditions as well as obesity and diabetic complications (4, 6–8). Further studies on somatostatinsecreting cells, and in particular, the mechanisms regulating somatostatin release, hold similar promise. Gastric D cells serve as a major source for circulating somatostatin and are of particular interest because of their relatively large numbers as compared with other sources and due to their distinctive ability to secrete somatostatin in response to a plethora of neurotransmitters, hormones as well as nutrients. These somatostatin-secreting cells are similar to other gastric endocrine cells, which include ghrelin cells, serotonin-secreting enterochromaffin cells, histamine-secreting enterochromaffin-like cells and gastrin-secreting G cells, in that they are sparsely distributed within the mucosa. Nestled among neighboring endocrine and exocrine cells, the D cells exert inhibitory input onto their neighbors’ secretory capacities via release of somatostatin from characteristic basal cytoplasmic peduncles. Gastric D cells are also major contributors to the picomolar concentrations of somatostatin found in circulation, even though most of the somatostatin produced in the stomach acts locally and is rapidly degraded by proteases. Somatostatin therefore lacks selectivity and precision in its role as a “hormone,” in terms of both site of origin and its target organs (9). The speckled distribution of D cells within the gastric mucosa has hindered the derivation of homogenous populations of D cells thus slowing research on their physiology. Also, lack of selective antibodies to membrane-bound G protein-coupled receptors (GPCRs) and other receptor classes has largely prevented immunohistochemical characterization of the signaling systems that directly regulate somatostatin secretion and other aspects of D-cell function. Instead, insights into somatostatin secretion from D cells so far have been obtained mostly

The Homeodomain-Containing Transcription Factors Arx and Pax4 Control Enteroendocrine Subtype Specification in Mice

Intestinal hormones are key regulators of digestion and energy homeostasis secreted by rare enteroendocrine cells. These cells produce over ten different hormones including GLP-1 and GIP peptides known to promote insulin secretion. To date, the molecular mechanisms controlling the specification of the various enteroendocrine subtypes from multipotent Neurog3+ endocrine progenitor cells, as well as their number, remain largely unknown. In contrast, in the embryonic pancreas, the opposite activities of Arx and Pax4 homeodomain transcription factors promote islet progenitor cells towards the different endocrine cell fates. In this study, we thus investigated the role of Arx and Pax4 in enteroendocrine subtype specification. The small intestine and colon of Arx- and Pax4-deficient mice were analyzed using histological, molecular, and lineage tracing approaches. We show that Arx is expressed in endocrine progenitors (Neurog3+) and in early differentiating (ChromograninA−) GLP-1-, GIP-, CCK-, Sct- Gastrin- and Ghrelin-producing cells. We noted a dramatic reduction or a complete loss of all these enteroendocrine cell types in Arx mutants. Serotonin- and Somatostatin-secreting cells do not express Arx and, accordingly, the differentiation of Serotonin cells was not affected in Arx mutants. However, the number of Somatostatin-expressing D-cells is increased as Arx-deficient progenitor cells are redirected to the D-cell lineage. In Pax4-deficient mice, the differentiation of Serotonin and Somatostatin cells is impaired, as well as of GIP and Gastrin cells. In contrast, the number of GLP-1 producing L-cells is increased concomitantly with an upregulation of Arx. Thus, while Arx and Pax4 are necessary for the development of L- and D-cells respectively, they conversely restrict D- and L-cells fates suggesting antagonistic functions in D/L cell allocation. In conclusion, these finding demonstrate that, downstream of Neurog3, the specification of a subset of enteroendocrine subtypes relies on both Arx and Pax4, while others depend only on Arx or Pax4.

Expression and cellular localization of somatostatin fromParalichthys olivaceus

Somatostatin(SS) called also somatotropin release-inhibiting factor is a peptide named for its ability to inhibit pituitary growth hormone release.To facilitate investigation of the SS function in the teleost,its coding sequence in paralichthys olivaceus was isolated by the technique of Rapid Amplification of cDNA Ends(RACE).SYBR Green I Quantitative Real-time PCR analysis and immunohistochemistry were used to assess the expression of SS at mRNA and protein level.The deduced protein sequence con-tains 126 amino acid residues and a predicted molecular mass of 14.3 kD.Very low level of SS mRNA is presented during the first three days after hatching.The expression level of the SS gene was increased con-stantly before metamorphosis and steadily during the metamorphosis,then increased significantly during post-metamorphosis.There was no SS-positive reaction in the oropharyngeal cavity and esophagus.SS secreting cells are only distributed in the stomach,pancreas and pituitary.Several types of SS secreting cells could be distinguished in above organs.In stomach,SS secreting cells are dispersively distributed among the epithelium cells,presenting various kinds of forms such as ovoid,globular,triangle,elongated or spindle-shaped aspects.The pancreas and liver of flounder combine into a single organ named hepato-pancreas,and SS-positive cells are only located in the pancreas,showing as globular-shaped,most of them are closed to blood vessels.SS-positive cells at high density are located in pars distalis of pituitary,so it is difficult to distinguish the shape of those cells.

Immunohistochemical localization of somatostatin and pancreatic polypeptide in smokers with chronic pancreatitis

Recent studies have demonstrated a significant role of tobacco smoking in the development of chronic pancreatitis. Although there are published papers on the effects of cigarette smoking on insulin secretion in patients, no data are available on the effects of smoking on pancreatic endocrine cells secreting somatostatin and pancreatic polypeptide. The aim of the study was to evaluate the effects of cigarette smoking on endocrine pancreatic function by immunolocalization of somatostatin and pancreatic polypeptide in the pancreas from smokers and non-smoking patients with chronic pancreatitis in comparison with healthy controls. The LSAB2-HRP technique with polyclonal antibodies was used for the immunolocalization of somatostatin and pancreatic polypeptide in histological preparations of the pancreas. The intensity of immunohistochemical reaction was calculated with digital image analysis. The study demonstrated increased numbers of somatostatin (D) secreting cells and pancreatic polypeptide (PP) cells and their altered location in pancreatic islets and parenchyma of smoking patients with chronic pancreatitis, as compared to non-smoking patients and healthy controls. Smoking patients showed significantly higher immunostaining of the hormones in the pancreas compared to non-smoking patients and healthy persons. This study indicates that smoking may play a significant role in the development of endocrine disturbances in the development of chronic pancreatitis.

Manserin, a secretogranin II-derived peptide, distributes in the rat endocrine pancreas colocalized with islet-cell specific manner

Manserin is a recently characterized 40-amino acid neuropeptide derived from secretogranin II, a protein belonging to the chromogranin family. Although the physiological roles of manserin have not been elucidated to date, manserin has been shown to distribute in not only the brain but also the endocrine system such as the pituitary and adrenal glands, suggesting its role in the endocrine system. The present study aimed to explore the occurrence and distribution of manserin in the rat pancreas using an immunohistochemical technique with a polyclonal antibody against rat manserin. Immunoreactivity for manserin was readily detected in almost whole islets of Langerhans whereas not at all in the exocrine pancreas. Manserin-expressing cells were not colocalized with the glucagon-secreting cells (alpha cells), whereas they colocalized with insulin-secreting cells (beta cells) and somatostatin-secreting cells (delta cells), although their intracellular distribution was different. These results indicate that manserin, occurring in the endocrine pancreas, may have a potential role in the endocrine system.

G and D cells in rat antral mucosa: an immunoelectron microscopic study.

To investigate the gastrin secreting cells (G cells) and the somatostatin secreting cells (D cells) of antral mucosa in rats at the ultrastructural level. Revised immunoelectron microscopic technique was used to detect the G cells and D cells in rat antral mucosa through gastrin and somatostatin antibodies labeled by colloidal gold. Also the relevant quantitative analysis regarding the granular number of colloidal gold in G cells and in D cells was conducted. Immunological granules of colloidal gold were distributed in G cells and D cells. Gastrin labeled golden granules or somatostatin labeled ones presented mainly as lobation-like or island-like congeries. Most of the golden congeries were observed dissociated in cytoplasms of G cells or D cells, near the basement membrane. A few golden congeries were located in nuclei. The number of golden granules in one G cell was around 107.04 +/- 19.68 and was 83.36 +/- 17.58 in one D cell. Gastrin secreting granules are located in cytoplasms and nuclei of G cells, and somatostatin secreting granules both in cytoplasms and in nuclei of D cells. The number of golden granules can be quantitatively analyzed to determine the relative amount of gastrin secreting granules or somatostatin secreting granules.

Localization of alpha-endosulphine in pancreatic somatostatin delta cells and expression during rat pancreas development.

alpha-Endosulphine, a protein that belongs to the cAMP-regulated-phosphoprotein family, has been reported to modulate insulin secretion in vitro through interaction with the pancreatic beta-cell ATP-sensitive potassium (K(ATP)) channel. In this study, we analysed the tissue distribution of alpha-endosulphine and determined its pancreatic cellular localization. Quantitative tissue distribution of alpha-endosulphine was studied by RIA on tissue extracts and cellular/subcellular localization was done using immunocytochemistry, morphometry and western blot analysis. alpha-Endosulphine and somatostatin release from RINT-3 somatostatin-secreting cells was quantified by RIA. alpha-Endosulphine, concentrated particularly in the central nervous system, was also detected in a wide variety of tissues including the pancreas. Immunohistochemistry analysis of adult rat pancreatic sections showed that alpha-endosulphine localized in somatostatin delta cells, where its expression increased during post-natal development. Immunoreactive cells were detected from foetal age E19, and the number of somatostatin cells co-expressing alpha-endosulphine increased with developmental age from E19 until adult. alpha-Endosulphine, highly expressed in the cytoplasm of RINT3 somatostatin-secreting cell line, was recovered in the particulate fraction of RINT3 cell extracts but was not co-secreted with somatostatin. alpha-Endosulphine is expressed in all tissues tested including pancreas and is also detected in plasma. Pancreatic alpha-endosulphine is specifically localized in somatostatin delta cells. This cytosolic protein is not co-secreted with somatostatin and could be physically associated with particulate components of the cells. These findings are not in favour of an endocrine/paracrine effect of alpha-endosulphine on the beta-cell K(ATP) channel.

Immunohistochemical localization of cellubrevin on secretory granules in pancreatic B-cells.

Cellubrevin is one of the proteins involved in the docking and fusion of secretory granules to the plasma membrane. It has been reported that cellubrevin is widely distributed in both neural and non-neural cells, including insulin-secreting B-cells. This study aims to demonstrate by immunohistochemical techniques that cellubrevin is localized in insulin-secreting cells and further to examine whether it might occur in glucagon- and somatostatin-secreting cells in the pancreatic islet in the rat and mouse. We used the polyclonal antibody against the N-terminal peptide whose specificity was confirmed by Western blot analysis. Double immuno-staining demonstrated that cellubrevin was localized in insulin-containing cells, but both glucagon-containing and somatostatin-containing cells lacked the immuno-reactivity. Immuno-electron microscopic analysis revealed the localization of cellubrevin on the margin of secretory granules near the plasma membrane but not in the granules closer to the nucleus. These observations support the view that cellubrevin in the pancreatic islet is expressed on the membrane of the secretory granules in B-cells at the stage of exocytosis.

Cholecystokinin, neuropeptide Y and galanin modulate the release of pancreatic somatostatin-25 and somatostatin- 14 in vitro

The direct effects of cholecystokinin (CCK), neuropeptide Y (NPY) and galanin on somatostatin-14 (SS-14) and somatostatin-25 (SS-25), containing [Tyr 7, Gly 10]-SS-14 at its carboxy terminus and presumably derived from a different gene than that giving rise to SS-14, were evaluated on principal islets (Brockmann bodies) removed from rainbow trout, Oncorhynchus mykiss. The potential for peptidergic action was deduced from the extensive neural network innervating the Brockmann body and confirmed by immunohistochemistry. Immunopositive CCK-like staining was located around the periphery of principal islets as well as in centrally-located nerve tracts. Mammalian CCK-33 stimulated SS-25 release at a dose of 10 −11 M in the presence of 1, 3 and 10 mM glucose. CCK-33 inhibited the release of SS-14 at concentrations of 10 −11 and 10 −7 M only in the presence of high (10 mM) glucose. Mammalian NPY stimulated SS-25 release in a dose-related manner in the presence of low (1 mM) glucose; the efficacy of this effect decreased at higher glucose concentrations. SS-14 release was inhibited by NPY in the presence of 10 mM glucose. Mammalian galanin generally inhibited the secretion of SS-25, but stimulated the release of SS-14 at 10 −7 M in the presence of 3 mM glucose. These results suggest that CCK, NPY and galanin modulate the release of the somatostatins present within the trout pancreas. Furthermore, glucose may mediate the overall responsiveness of somatostatin-secreting cells to these peptides.

Functional active receptors for insulin-like growth factor-I (IGF-I) and IGF-II on insulin-, glucagon-, and somatostatin-producing cells

Insulin-like growth factors I and II (IGF-I and IGF-II) are expressed at high levels in the endocrine pancreas during development and tissue regeneration. However, their effects at the endocrine pancreas are poorly understood. We searched for receptors of IGF-I and IGF-II and possible biological effects on clonal insulin-secreting (HIT), glucagon-secreting (INR1G9), and somatostatin-secreting (RIN 1027 B2) cell lines. Our data showed that HIT cells and RIN 1027 B2 cells express specific type I and type II IGF receptors. INR1G9 cells possess type II IGF receptors and IGF-I binding sites with the same affinity for both IGF-I and IGF-II. In HIT cells, insulin secretion was not influenced by either peptide. Proinsulin gene transcription was stimulated by IGF-II but not by IGF-I. IGF-I potently inhibited proglucagon gene transcription and glucagon secretion in INR1G9 cells, whereas IGF-II only inhibited glucagon release. In RIN 1027 B2 cells, IGF-I but not IGF-II increased somatostatin output, whereas both stimulated somatostatin gene expression. These data demonstrate the presence of classic type I and type II IGF receptors on insulin-, glucagon-, and somatostatin-secreting cells. Both peptides may be important regulators of endocrine pancreatic function in terms of islet hormone release and gene expression. Therefore, both peptides may be involved in the regulation of intraislet cellular homeostasis.

Modulation of cytoplasmic Ca2+ signals in somatostatin-secreting cells (QGP-1N) by glucose.

Modulation of cytoplasmic Ca2+ signals in somatostatin-secreting cells (QGP-1N) by glucose.

Intracellular Ca2+ signals in human-derived pancreatic somatostatin-secreting cells (QGP-1N).

Single-cell microfluorimetry techniques have been used to examine the effects of acetylcholine (0.1-100 microM) on the intracellular free calcium ion concentration ([Ca2+]i) in a human-derived pancreatic somatostatin-secreting cell line, QGP-1N. When applied to the bath solution, acetylcholine was found to evoke a marked and rapid increase in [Ca2+]i at all concentrations tested. These responses were either sustained, or associated with the generation of complex patterns of [Ca2+]i transients. Overall, the pattern of response was concentration related. In general, 0.1-10 microM acetylcholine initiated a series of repetitive oscillations in cytoplasmic Ca2+, whilst at higher concentrations the responses consisted of a rapid rise in [Ca2+]i followed by a smaller more sustained increase. Without external Ca2+, 100 microM acetylcholine caused only a transient rise in [Ca2+]i, whereas lower concentrations of the agonist were able to initiate, but not maintain, [Ca2+]i oscillations. Acetylcholine-evoked Ca2+ signals were abolished by atropine (1-10 microM), verapamil (100 microM) and caffeine (20 mM). Nifedipine failed to have any significant effect upon agonist-evoked increases in [Ca2+]i, whilst 50 mM KCl, used to depolarise the cell membrane, only elicited a transient increase in [Ca2+]i. Ryanodine (50-500 nM) and caffeine (1-20 mM) did not increase basal Ca2+ levels, but the Ca(2+)-ATPase inhibitors 2,5-di(tert-butyl)-hydroquinone (TBQ) and thapsigargin both elevated [Ca2+]i levels. These data demonstrate for the first time cytosolic Ca2+ signals in single isolated somatostatin-secreting cells of the pancreas. We have demonstrated that acetylcholine will evoke both Ca2+ influx and Ca2+ mobilisation, and we have partially addressed the subcellular mechanism responsible for these events.

Elevation of intracellular calcium signals in single isolated somatostatin-secreting cells derived from a pancreatic endocrine tumour.

Elevation of intracellular calcium signals in single isolated somatostatin-secreting cells derived from a pancreatic endocrine tumour.

Heterogeneous expression of glucokinase among pancreatic beta cells.

The cellular location of glucokinase (GK), a key component of the glucose-sensing mechanism of the pancreatic islet, was determined using immunocytochemical techniques. In rat islets, GK immunoreactivity was detected only in beta cells with no immunoreactivity detected in alpha, delta, or pancreatic polypeptide-containing (PP) cells. However, within various beta cells, GK immunoreactivity varied considerably. Most beta cells displayed relatively low levels of cytoplasmic immunoreactivity whereas other beta cells stained intensely for this enzyme. Colocalization studies of GK and GLUT2, the high Km glucose transporter of beta cells, confirmed that these proteins are located in different subcellular domains of beta cells. The lack of GK immunoreactivity in glucagon- and somatostatin-secreting cells in islets suggests that these cells are not directly responsive to glucose or utilize a fundamentally different mechanism for sensing glucose fluctuations. Moreover, the differential expression of GK among pancreatic beta cells suggests that glucose phosphorylation is the probable enzymatic control point for the functional diversity of these cells.

Insulin suppression is associated with hypersomatostatinemia and hyperglucagonemia in glucose-injected rainbow trout

Rainbow trout, Oncorhynchus mykiss, were used to evaluate the effects of carbohydrate loading on plasma levels of pancreatic hormones and associated changes in metabolic indexes in a carnivorous fish. Glucose (3,000 mg/dl, 10 microliters/g body wt) was injected intraperitoneally into fish (mean wt 54 +/- 5 g) that were killed 0.5-24 h after administration. Glucose injection resulted in hyperglycemia with maximum glucose levels of 306 +/- 13 mg/dl observed 60 min after injection. Glucose administration also resulted in hyperlipidemia. Plasma fatty acids increased twofold in glucose-injected animals. Alterations in plasma metabolites reflected changes in energy stores. Although total lipid concentration was unaffected by glucose injection, lipolytic enzyme activity in the liver was enhanced. Biosynthetic capacity, as indicated by NADPH production from glucose-6-phosphate dehydrogenase, was decreased by glucose injection. Liver glycogen content was reduced in glucose-injected animals 1 h after injection. Glucose injection was attended by increases in the plasma levels of gene II somatostatin-25 (predominant form of pancreatic somatostatin in salmonids) and of glucagon. Insulin levels were initially suppressed after glucose injection. These results indicate that metabolic adjustments caused by glucose administration can be related to the regulatory action of pancreatic hormones. Furthermore, these results suggest that the somatostatin-secreting cells of the trout are sensitive to glucose and that somatostatin-suppressed insulin secretion contributes to the glucose intolerance of trout.

P 2 purinoceptor agonists stimulate somatostatin secretion from dog pancreas

The effects of ATP and ADP structural analogues (2-methylthio ATP; α,ß-methylene ADP) on somatostatin secretion were tested in dogs. Insulin and glucagon secretion was also evaluated. Our experiments were performed in vivo and vitro. In vivo, 2-methylthio ATP was infused directly into the pancreaticoduodenal artery of anesthetized dogs and blood was sampled from the pancreaticoduodenal vein. This ATP analogue (≈ 15 μM) immediately induced stimulation of both somatostatin and insulin secretion, which was accompanied by a slight reduction of glycemia. A delayed increase in glucagon output was observed. In vitro, using the isolated perfused dog pancreas uncinate process, α,ß-methylene ADP, a stable ADP analogue (16.5 μM), was infused in the presence of a substimulating glucose concentration (4.2 mM). Under these conditions, α,ß-methylene ADP immediately induced the stimulation of somatostatin secretion without affecting basal insulin and glucagon secretion. In conclusion our results suggest the presence of P 2 purinoceptors on pancreatic somatostatin secreting cells.

Morphogenesis and distribution of the endocrine pancreas in adult lampreys

The endocrine pancreas of larval lampreys appears as islets of cells isolated in the submucosa and those both continuous with, and within, the gut epithelium at the intestinal-oesophageal-bile duct junction. The islets, and occasionally follicles, are composed of only insulin-secreting (B) cells. During metamorphosis, the bile duct either completely degenerates or its epithelium transforms into a caudal endocrine pancreas while a cranial pancreas appears as a specialization and expansion of the larval pancreas. Immunocytochemistry and histochemistry demonstrates that there is a wide variation in the distribution of the pancreatic tissue in adults of lamprey species, and this variation may result from interspecific differences in morphogenetic events at metamorphosis. Despite species variability in its distribution, the endocrine pancreatic tissue in all adult lampreys is composed of equal numbers of B cells and somatostatin-secreting (D) cells, but there are no glucagon-secreting (A) cells. Immunocytochemistry reveals that B and D cells of the caudal pancreas differentiate from cells of the larval bile duct during metamorphosis of the sea lamprey,Petromyzon marinus.