Ontogeny of the endocrine pancreas in sea bass (Dicentrarchus labrax): an ultrastructural study. II. The big and secondary islets

The big and secondary islets of sea bass larvae were characterized ultrastructurally from, 25 to 60 days after hatching. From the 25th day, big islets consisted of inner type II and III, external type I and peripheral type IV cells. From the 55th day, type V cells appeared in limited peripheral areas. Secondary islets, first found in 32-day-old larvae, were made up of inner type II and III, external type I, and peripheral either type IV and V cells (type I islets), or only type V cells (type II islets). Type I cells contained secretory granules with a fine granular, low-medium electron-dense material, whereas the secretory granules of type II cells were smaller and had a high electron-dense core with diffused limits; needle and rod-like crystalloid contents were occasionally found. Type III secretory granules posessed a homogeneous, high or medium electron-dense material with or without a clear halo. Type IV cells had secretory granules with a polygonal dense core embedded in a granular matrix and granules containing a high or medium electron-dense material. Type V cells had secretory granules with a fine granular, high or medium electron-dense content. These cell-types correlated with cells previously identified immuno-cytochemically, as regards to their distribution in the islets, and related to those characterized ultrastructurally in adult specimens. Thus, types I, II, III, IV and V correspond to D₁, B, D₂, A and PP cells, respectively. From the 32nd day onwards, endocrine cells of all the different types were found grouped, type V cells also being observed in isolation close to pancreatic ducts and/or blood vessels. Small groups consisting of type I and II cells were found in 40-day-old larvae. A mitotic centroacinar ductular cell containing some secretory granules similar to those of type I cells, was seen adjacent to a type I cell. As the larvae grew older, the endoplasmic reticulum developed, the number of free ribosomes decreased, and the number and size of the secretory granules increased. Dark type I, II, III, IV and V cells were found in the islets and cell clusters from the 55th day onwards.     

Reg genes are CCK2 receptor targets in ElasCCK2 mice pancreas

We previously demonstrated that expression of the gastrin receptor, CCK2R, in pancreatic acini of transgenic ElasCCK2 mice induced alteration of acinar morphology and differentiation, increased sensitivity to a carcinogen and development of preneoplastic lesions and tumours. Reg proteins are suggested to be involved in pancreatic cancer and in regeneration of endocrine pancreas. Reg I gene is a known target of gastrin. We examined whether an expression of CCK2R in the pancreatic acini of ElasCCK2 mice is linked to induction of Reg proteins expression. We analyzed Reg expression by Western-blot and immunohistochemistry in pancreas from ElasCCK2 and control mice. Islet neogenesis, glucose homeostasis, insulin secretion and content were also evaluated. Reg I is exclusively produced in acini in ElasCCK2 and control mice. In tumoral pancreas, Reg I and Reg III proteins are expressed in duct-like cells in preneoplastic lesions or in the periphery of tumours and in adjacent acini. The expression of Reg III proteins is increased in ElasCCK2 pancreas before the development of preneoplastic lesions in a subpopulation of islet cells and in small islet-like cell clusters dispersed within the acinar tissue. Several criteria of an enhanced neogenesis are fulfilled in ElasCCK2 pancreas. Moreover, ElasCCK2 mice have an improved response to glucose load, an increased insulin secretion and a doubling of insulin content compared to control mice. We show that Reg proteins are targets of CCK2R activation and are induced during early steps of carcinogenesis in ElasCCK2 mice pancreas. Alterations of exocrine tissue homeostasis in ElasCCK2 pancreas concomitantly activate regenerative responses of the endocrine pancreas possibly linked to paracrine actions of Reg III proteins.     

Immunocytochemical characterization of glucagon-immunoreactive cells using monoclonal antibodies to pancreatic glucagon

Monoclonal antibodies raised to pancreatic glucagon were tested for their ability to detect glucagon-containing endocrine cells in material processed for light and electron microscopy. Samples from man, baboon and rat were used in this investigation. Two antibodies were specific for the pancreatic islet A cells, the remainder detected both pancreatic and enteric endocrine cells. In man and baboon the glucagon-containing cells were confined to the pancreas, lower small intestine and colon. In the rat the distribution was extended to include the corpus of the stomach and the jejunum. The cells identified in the ileum and colon were of three morphological types endocrine, paracrine (type 1) with a single basal process and paracrine (type 2) with multiple small cytoplasmic processes. These antibodies also detected cells in material fixed by conventional methods for electron microscopy. The ultrastructural appearance of the baboon pancreatic glucagon-containing ultracellular secretory granules were demonstrated to be clearly distinct from those described previously in man and rat. The secretory granules averaged 330 +/- 23 nm and lacked the characteristic clear outer halo seen in the other two species.     

Ultrastructural identification of a new cell type — the N-cell as the source of neurotensin in the gut mucosa

The neurotensin-cell is identified immunohistochemically and ultrastructurally by differential counting of endocrine cells in the gut of a primate (Tupaia belangeri). Utilizing light microscopy, the EC-cells are identified by the Masson-Fontana silver stain; with the same method the neurotensin cells are not stained. The other endocrine cells have been quantified in the small intestine using the peroxidase-antiperoxidase stain with antisera against glucagon, somatostatin, cholecystokinin, gastrin, secretin, pancreatic polypeptide, gastric inhibitory peptide and neurotensin. In the ileal mucosa of Tupaia, the most frequent endocrine cell is the EC-cell followed by the glucagonoid cell, (L-cell). The immunoreactive neurotensin cell represents the third most frequent endocrine cell in this region. On the ultrastructural level, this third most frequent endocrine cell is a heretofore undescribed cell, the N-cell, containing electron dense secretory granules measuring 335 +/- 87 nm in diameter.     

An immunocytochemical study of the endocrine pancreas in the Australian fat-tailed dunnart (Sminthopsis crassicaudata)

Summary The endocrine pancreas of the Australian fattailed dunnart, Sminthopsis crassicaudata, was investigated by means of electron-microscopic immunocytochemistry using the protein A-gold technique on London resin (LR) white-embedded tissue. The primary antibodies used were raised against insulin, glucagon, somatostatin and pancreatic polypeptide. The morphology of the secretory granules differed in the four cell types. The insulin cells are pleomorphic, and the secretory granules composed of an electron-dense core surrounded by an electron-lucen halo. The glucago cells possess granules with an electron-dense core usually surrounded by a halo of less dense granular material. Somatostatin cells have large, less dense secretory granules. The pancreatic polypeptide cells show small, dense secretory granules. In order for an ultrastructural study to be considered reliable for the definite identification of endocrine cell types, it is essential that it be corroborated by immunocytochemical data at the light-or preferably electron-microscopic level. Recent developments in immuno-electron-microscopic techniques have contributed to a better knowledge of cells responsible for the secretion of a wide variety of hormones, as in this study.     

Immunohistochemical and ultrastructural study of anterior pituitary cells in the female Afghan pika,Ochotona rufescens rufescens

An immunohistochemical study of the anterior pituitary gland of the female Afghan pika was carried out to distinguish the ultrastructural features of GH, PRL, ACTH, TSH and LH cells. The histochemically identified GH cells resembled ultrastructurally oval or round GH cells of the rat laden with large, dense secretory granules. PRL cells were divided into three subtypes based on differences in the diameter of their spherical secretory granules. They lacked polymorphic or irregularly shaped secretory granules. ACTH cells resembled ultrastructurally, in some respects, Siperstein's "corticotrophs" of the rat with peripheral arrangement of secretory granules. However, they were not always stellate, but elongate or angular in shape. The dense secretory granules were concentrated in the peripheral area of cytoplasm. TSH cells were non-stellate, but usually oval in shape, containing the smallest spherical secretory granules (100-200 nm in diameter). Almost all LH cells reacted also with FSH antiserum. They were irregular in shape, sometimes in contact with or surrounded the GH cells. They contained an abundance of medium-sized secretory granules (140-260 nm in diameter) which were larger than those in the LH cells of the female rat throughout the estrous cycle. Large secretory granules in the LH cells of the female pika seemed to be related to the endocrine state of persistent estrus.     

Immunocytochemical characterization of glucagon-immunoreactive cells using monoclonal antibodies to pancreatic glucagon

Summary Monoclonal antibodies raised to pancreatic glucagon were tested for their ability to detect glucagon-containing endocrine cells in material processed for light and electron microscopy. Samples from man, baboon and rat were used in this investigation. Two antibodies were specific for the pancreatic islet A cells, the remainder detected both pancreatic and enteric endocrine cells.In man and baboon the glucagon-containing cells were confined to the pancreas, lower small intestine and colon. In the rat the distribution was extended to include the corpus of the stomach and the jejunum. The cells identified in the ileum and colon were of three morphological types endocrine, paracrine (type 1) with a single basal process and paracrine (type 2) with multiple small cytoplasmic processes.These antibodies also detected cells in material fixed by conventional methods for electron microscopy. The ultrastructural appearance of the baboon pancreatic glucagon-containing ultracellular secretory granules were demonstrated to be clearly distinct from those described previously in man and rat. The secretory granules averaged 330±23 nm and lacked the characteristic clear outer halo seen in the other two species.     

Ultrastructure of endocrine pancreatic granules during pancreatic differentiation in the grass snake,Natrix natrix L. (Lepidosauria,Serpentes)

We used transmission electron microscopy to study the pancreatic main endocrine cell types in the embryos of the grass snake Natrix natrix L. with focus on the morphology of their secretory granules. The embryonic endocrine part of the pancreas in the grass snake contains four main types of cells (A, B, D, and PP), which is similar to other vertebrates. The B granules contained a moderately electron‐dense crystalline‐like core that was polygonal in shape and an electron‐dense outer zone. The A granules had a spherical electron‐dense eccentrically located core and a moderately electron‐dense outer zone. The D granules were filled with a moderately electron‐dense non‐homogeneous content. The PP granules had a spherical electron‐dense core with an electron translucent outer zone. Within the main types of granules (A, B, D, PP), different morphological subtypes were recognized that indicated their maturity, which may be related to the different content of these granules during the process of maturation. The sequence of pancreatic endocrine cell differentiation in grass snake embryos differs from that in many vertebrates. In the grass snake embryos, the B and D cells differentiated earlier than A and PP cells. The different sequence of endocrine cell differentiation in snakes and other vertebrates has been related to phylogenetic position and nutrition during early developmental stages.     

The WDHA syndrome: clinical and laboratory data on 28 Japanese cases

During the period between 1967 and 1983, there were 28 WDHA patients in Japan. Clinically, these patients were not very different from those reported in western countries, but we have been able to add a few observations as follows. First, the WDHA patients with pancreatic endocrine tumors and those with neuroblastic tumors were present in almost equal numbers. Second, fluctuating hypercalcemia was often observed in patients with this syndrome, but it was not observed in any watery diarrhea due to the other etiologies. Third, the WDHA patients who have multiple endocrine neoplasia (MEN) type 1 have multiple tumors in the pancreas, indicating that the patients with MEN should be managed in a different manner in regard to determining tumor location as well as applying surgical treatment. When various types of tumors were examined for VIP content, it was found that many tumors have the potential to produce VIP, and when VIP is produced in large amounts, plasma VIP levels become elevated, resulting in the WDHA syndrome.     

Ontogeny of the endocrine pancreas in sea bass (Dicentrarchus labrax): an ultrastructural study. I. The primordial cord and the primitive,single and primordial islets

The primordial cord and the primitive, single and primordial islets present in the 3 earliest stages of the developing endocrine pancreas of sea bass were studied ultrastructurally. The primordial cord consisted of type I and II cells and was included in the gut. Besides these cell types, X cells were seen in the primitive islet. The single islet was made up of type I, II, III and IV cells. A correlation between these endocrine cell-types and cells previously identified immunocytochemically, was established. Type I, II, III and IV cells, correlated respectively with SST-25-, insulin-, SST-14- and glucagon-immunoreactive cells, and could be related to the D₁, B, D₂ and A cells, respectively, of older larvae and adult sea bass. Each cell type shows characteristic secretory granules from its first appearance. A progressive development of the organelles and an increase in the number and size of the secretory granules, whose ultrastructure also varied, was observed in the endocrine cells of the primordial cord and the succeeding islets. In 25-day-old larvae at the beginning of the fourth developmental stage, the primordial islet, the first ventral islet found, was close to a pancreatic duct and blood vessel, and consisted of type I and II cells whose ultrastructure was similar to that of the type I and II cells in the primordial cord. These data suggest a ductular origin for the pancreatic endocrine cells in the ventral pancreas. It is suggested that although endocrine cells undergo mitosis, their increase in number during the earliest development stages is principally due to the differentiation of surrounding cells.     

Identification of glucagon-producing cells (A cells) in dog gastric mucosa

An immunocytochemical technique using specific antiglucagon serum reveals the presence of glucagon-containing cells situated exclusively in the oxyntic glandular mucosa of the dog stomach. Electron microscope examination of the mucosa demonstrated endocrine cells containing secretory granules with a round dense core surrounded by a clear halo, indistinguishable from secretory granules of pancreatic A cells. Like the alpha granules of pancreatic A cells, the granules of these gastric endocrine cells exhibited a peripheral distribution of silver grains after Grimelius silver staining. Moreover, the granules of these cells were found to be specifically labeled with reaction product, using the peroxidase immunocytochemical technique at the ultrastructural level. Accordingly, these cells were named gastric A cells. These data suggest that the gastric oxyntic mucosa contains cells indistinguishable cytologically, cytochemically, and immunocytochemically from pancreatic A cells. It is believed that gastric A cells are responsible for the secretion of the gastric glucagon.     

Immunocytochemical study of the endocrine pancreas in the grey kangaroo (Macropus fuliginosus)

Summary The endocrine pancreas of the grey kangaroo,Macropus fuliginosus, was investigated by means of immunocytochemistry using the PAP method on the same section at the light- and electron-microscopic levels. Semithin plastic sections were stained individually with primary antibodies for insulin, glucagon, somatostatin and pancreatic polypeptide (PP), and then photographed. Sections were osmicated, re-embedded in BEEM capsules, and ultrathin sections made and examined. The same labelled cells as in the semithin sections were localised in the thin sections, photographs taken and the morphology of secretory granules studied. The insulin cells were pleomorphic; their secretory granules displayed an electron-dense core surrounded by an empty halo. The glucagon cells possessed granules with an electron-dense core usually surrounded by a halo of less dense granular material. Somatostatin cells had larger, less dense secretory granules. The PP cells showed small, dense secretory granules. In order for an ultrastructural study to be considered reliable for the definite identification of endocrine cell types, it is essential that it be corroborted by correlated immunocytochemical data at the light-and electron-microscopic levels.     

Ultrastructural studies of the ontogeny of fetal human and porcine endocrine pancreas, with special reference to colocalization of the four major islet hormones.

Most, if not all, endocrine cells seem capable of synthesizing and storing more than one hormone. Such cellular colocalization of hormones can be due either to the presence of two or more specific granules within the cells or to colocalization of the hormones within a single granule. The present study was performed to clarify the subcellular localization of insulin, glucagon, somatostatin, and pancreatic polypeptide within the endocrine cells of the human and porcine pancreas during fetal development, with special reference to possible colocalization of the hormones. The tissue specimens were processed for ultrastructural cytochemistry using Lowicryl as embedding medium. An immunogold labeling technique was used with two parallel, but not interacting, antibody chains. Sections from each specimen were double labeled in different combinations giving a complete covering of the four major islet hormones. During fetal life (50-90 days prenatally in porcine pancreas, 14 weeks gestation in the human pancreas) several hormones were demonstrated, not only in the same endocrine cells, but also in the same secretory granules (polyhormonal granules). Costorage of insulin, glucagon, somatostatin, and pancreatic polypeptide was demonstrated in granules in pancreatic endocrine fetal cells. At an early fetal stage, the endocrine cells contained either dense, round granules or pale, heteromorphous granules. With increasing age and maturation of the endocrine cells, structural differentiation of the secretory granules was found to be associated with a gradual disappearance of the polyhormonal granules. The first genuine monohormonal cell to appear in the porcine fetus was the pancreatic polypeptide cell (at 70 days gestation); it was followed by the somatostatin-producing endocrine cell. Mature insulin- and glucagon-producing cells were only demonstrated after birth. Thus, in the adult pancreatic endocrine cells, each specific endocrine cell type produced only one of the four classical hormones. The present investigation demonstrated that the endocrine cells of the fetal, but not the adult, pancreas are able to synthesize all the major islet hormones, and that these peptides are costored in the same granule. The data obtained support the concept of a common precursor stem cell for pancreatic hormone-producing cells.     

A morphometric study of the secretory process in the endocrine pancreas of the foetal rat

The present study concerns pancreatic beta cells from rat foetus at 18, 19 and 21 days of gestation. On micrographs, the cytoplasm of beta cells was subdivided into 3 zones: one zone corresponding roughly to the cell web, a second zone just underlying the cell web, and a third zone comprising the remaining cytoplasm. The secretory granules present in each zone were counted; in the cell web, granules fused with plasma membrane were counted separately. During later foetal stages the increase in the frequency of granule to plasma membrane fusions parallels the increase in blood insulin levels, and the total number of granules in beta cells increases in parallel with the pancreatic insulin content. Therefore, as the beta cell matures, both secretion and biosynthesis of insulin increase sharply. The observed changes in the distribution of the granules in the different zones of the cytoplasm with the foetal age suggests that the cell web controls the access of the granules to the plasma membrane. The morphometric technique used allows a direct determination, at the cellular level, of even small variations in exocytosis-mediated secretory discharge and suggests a regulatory role of the cell web.     

Quantitative immunocytochemical analysis of the endocrine pancreas of the Nile crocodile

Four major pancreatic hormones were immunolocalized at the light and electron microscopic levels in the pancreas of the Nile crocodile, Crocodilus niloticus. Immunogold was used for electron microscopy, and peroxidase-antiperoxidase was used for light microscopy. Somatostatin-positive D-cells and pancreatic polypeptide-containing F-cells accounted for about 60% of the immunoreactive cells in the ventral pancreas. Glucagon-positive A-cells were the least frequent cell type in the ventral pancreas, about 15%, but were the predominant cell type, about 40%, in the pancreas that was dorsal in character. An expanded population of D-cells (relative to mammals and other higher vertebrates) in association with two very different numbers of A-cells can be expected to have important consequences for the homotropic control of secretory activity of the endocrine pancreas as well as for the function of the acinar pancreas. F-cells were absent from the dorsal part of the pancreas, whereas insulin-containing B-cells were slightly more abundant in this portion of the pancreas. The regional character of the endocrine pancreas was related to the complex looping of the proximal small intestine. Without immunolabeling, only B-granules were morphognomonic in electron micrographs. The insulin-reactive B-granules were the smallest (370 nm) of the secretory granules and were followed in size by somatostatin-positive D-granules (380 nm). The pancreatic polypeptide-containing secretory granules were the largest (580 nm). Glucagon-reactive A-granules (430 nm) sometimes exhibited a protuberance or extension of secretory granule matrix and limiting membrane. Such a morphological feature has previously been associated with secretion of glucagon and the initiation of insulin secretion. Taken together these studies indicate that protuberances have a significant, but as yet undefined, role in pancreatic endocrine cells.     

Ultrastructural and Fluorescence Microscopical Characterization of the Intestinal Endocrine Cells in a Cyclostome,Myxine glutinosa

Endocrine cells of so-called basal-granulated-open type in the intestinal epithelium of a cyclostome, the Atlantic hagfish (Myxine glutinosa), are characterized ultrastructurally and fluorescence microscopically. These cells regularly extend from the basal lamina to the gut lumen, ending in an apical process with microvilli and a filamentous surface coat. Fasting results in an accumulation of secretion granules in all cytoplasmic portions, except for the terminal web area. A similarity is recorded between the distribution of secretion granules and the finely granular fluorescamine-induced fluorescence, suggesting that the fluorescence is associated to some component(s) of the secretory granules. Granule release may take place at the base after an adequate stimulus (presence of food?) at the luminal portion of the cells. The formaldehyde condensation technique shows that insulin-containing hagfish islet parenchymal cells, but not intestinal endocrine cells, store dopamine after intestinal supply of the amine precursor. Acidification of formaldehyde vapour-fixed intestinal epithelium induces fluorescence in the granules of zymogen cells but not of endocrine cells, indicating a low concentration of tryptophyl-peptide(s) in the secretory granules of hagfish intestinal endocrine cells.     

Embryogenesis of the murine endocrine pancreas; early expression of pancreatic polypeptide gene.

By immunofluorescence on cytospin preparations and on semithin sections of mouse pancreatic buds, we have found glucagon and pancreatic polypeptide (PP)-containing cells at embryonal day 10.5 (E 10.5) in dorsal buds and at E 11.5 in ventral buds. Insulin-containing cells appear in dorsal buds at E 11.5, and one to two days later in ventral buds. Somatostatin-containing cells are detectable from E 13.5 in both dorsal and ventral buds. A quantitative analysis shows that up to E 15.5, PP-containing cells are relatively abundant in both buds. By PCR amplification of oligo(dT)-primed cDNAs prepared from total pancreatic RNA, we also detect PP mRNA from E 10.5 onwards, thus confirming the early expression of the PP gene in the developing mouse pancreas. Analysis of endocrine cells in situ suggests three major patterns of cell distribution in embryonic pancreas. First, individual hormone-containing cells are located within the epithelium of pancreatic ducts. In both dorsal and ventral buds, the majority of these endocrine cells contain PP, but many also contain glucagon, insulin or somatostatin. Secondly, clusters of endocrine cells are found in the pancreatic interstitium. Many of these cells contain both glucagon and PP which, by immunogold labelling of consecutive thin sections, can be shown to co-exist within individual secretory granules. Finally, starting on E 18.5, typical islets are formed with centrally located B cells and with the adult 'one cell-one hormone' phenotype. These results suggest an intriguing ontogenic relationship between A- and PP-cells, and also indicate that PP-containing cells may occupy a hitherto unexpected place in the lineage of endocrine islet cells.     

Ultrastructural localization of chromogranin: a potential marker for the electron microscopical recognition of endocrine cell secretory granules

Summary Using a monoclonal antibody (LK2H10) directed against human chromogranin, we have been able to localize this soluble glycoprotein to the matrix of secretory granules from a wide variety of endocrine cells. In the gut, enterochromaffin, enteroglucagon, glucose-dependent insulinotropic peptide, gastrin, and neurotensin-containing cells exhibit chromogranin immunoreactivity. In our system, chromogranin-immunoreactive material was restricted to the halo of human pancreatic glucagon-containing secretory granules within A-cells. Chromogranin immunoreactivity was also localized to secretory granules in phaeochromocytomas, gastrinomas, medullary carcinomas of the thyroid and a carotid body tumour (chemodectoma). Chromogranin is proposed as a potential marker for the ultrastructural recognition of endocrine cell secretory granules.     

Coexistence of pancreatic polypeptide (PP)-and glucagon-immunoreactivity in pancreatic endocrine cells of mouse

Immunocytochemical double staining techniques were used to study PP- and glucagon-like-immunoreactivity in pancreatic endocrine cells of mouse. An antiserum against FMRFamide appeared to react with all PP-immunoreactive endocrine cells. With fluorescence microscopy most PP/FMRFamide-immunoreactive cells also showed glucagon-immunoreactivity, but cells containing only PP- or glucagon-like substances were found as well. The proportion of cells containing PP-, glucagon, and both immunoreactivities varied strongly from islet to islet in all parts of the pancreas. Using an electron microscopical immunogold double staining procedure on Lowicryl-embedded pancreas, PP/FMRFamide- and glucagon-immunoreactivity appeared to be present in the majority of endocrine A cells; both immunoreactivities were randomly distributed within the granules of these cells. Cells containing only PP/FMRFamide- or glucagon-immunoreactivity were also found. Glucagon- and a faint FMRFamide-immunoreactivity was also observed in osmicated epon-embedded tissue. Independent of their immunoreactivity all positive cells showed the same round electron dense secretory granules.     

Localization of xenin-immunoreactive cells in the duodenal mucosa of humans and various mammals.

Xenin is a 25-amino-acid peptide extractable from mammalian tissue. This peptide is biologically active. It stimulates exocrine pancreatic secretion and intestinal motility and inhibits gastric secretion of acid and food intake. Xenin circulates in the human plasma after meals. In this study, the cellular origin of xenin in the gastro-entero-pancreatic system of humans, Rhesus monkeys, and dogs was investigated by immunohistochemistry and immunoelectron microscopy. Sequence-specific antibodies against xenin detected specific endocrine cells in the duodenal and jejunal mucosa of all three species. These xenin-immunoreactive cells were distinct from enterochromaffin, somatostatin, motilin, cholecystokinin, neurotensin, and secretin cells, and comprised 8.8% of the chromogranin A-positive cells in the dog duodenum and 4.6% of the chromogranin A-positive cells in human duodenum. In all three species, co-localization of xenin was found with a subpopulation of gastric inhibitory polypeptide (GIP)-immunoreactive cells. Immunoelectron microscopy in the canine duodenal mucosa demonstrated accumulation of gold particles in round, homogeneous, and osmiophilic secretory granules with a closely adhering membrane of 187 +/- 19 nm diameter (mean +/- SEM). This cell type was found to be identical to the previously described canine GIP cell. Immunocytochemical expression of the peptide xenin in a subpopulation of chromogranin A-positive cells as well as the localization of xenin immunoreactivity in ultrastructurally characterized secretory granules permitted the identification of a novel endocrine cell type as the cellular source of circulating xenin.