PubMed Health⌕ Search

Biomedical subjects

B Wiedenmann

Publications and source records attributed to B Wiedenmann.

At least 145 records · Page 8Linked to original sources
PubMed1994

Rat pancreatic AR42J cells. Amphicrine cells as an in vitro model to study peptide hormone receptor regulation.

The rat pancreatic acinar tumor cell line, AR42J, is widely used to study pancreatic acinar cell biology and biochemistry. In addition to the well-documented exocrine cell features, we have identified by immunofluorescence and by electron microscopy the co-expression of small neuroendocrine (NE) vesicles using the NE vesicle-specific markers synaptophysin and "protein S.V.2." AR24J cells store [3H]GABA, which is secreted upon potassium depolarization in a calcium-dependent manner. In addition, we found the expression of the receptor for the neurotransmitter substance P by using a receptor-specific cDNA probe. Glucocorticoid treatment, which profoundly inhibits cellular growth and induces differentiation, results in a rapid decrease of substance P receptor (SPR) gene expression as assessed by Northern blot analysis. Intracellular Ca2+ mobilization was then determined in response to substance P in control cells and glucocorticoid-pretreated cells by dual wavelength spectrophotometry using fura-2 in single cells. Glucocorticoid-mediated down-regulation of substance P receptors resulted in a dose- and time-dependent decrease of the intracellular Ca2+ mobilization stimulated by substance P. In summary, these data indicate that AR42J cells display an amphicrine phenotype with two differentially regulated secretory pathways; during glucocorticoid-induced differentiation, the cells become less sensitive to substance P stimulation as a consequence of reduced gene expression of the substance P receptor.

Animals↗
PubMed1994

Endosonography of neuroendocrine tumors of the stomach, duodenum, and pancreas.

Neuroendocrine tumors (NETs) of the foregut type are frequently smaller than 2 cm in diameter and mainly located in the pancreas or the gastric and duodenal wall. Conventional cross-sectional imaging techniques, such as transabdominal ultrasonography (US), computed tomography (CT), and magnetic resonance imaging (MRI) are limited by their inability to detect small tumors and especially those located within the gastrointestinal wall. Endoscopic ultrasonography (EUS) allows detailed visualization of the whole pancreas and almost all parts of the gastric and duodenal walls. Therefore, EUS is an important diagnostic tool for the preoperative localization of NETs of the foregut type. Several studies performed in a retrospective manner, as well as two studies performed in a prospective manner, indicate a clear superiority of EUS as compared to CT, US, MRI, and also angiography in detecting NETs of the foregut type. Somatostatin-receptor scintigraphy (SRS) also detects NETs of the foregut type in a very high percentage of cases, and the combination of EUS and SRS appears to increase the sensitivity even more. Thus EUS and also SRS should be employed early if NETs of the foregut type are suspected. Conventional imaging procedures such as US, CT, and MRI should be mainly used to exclude local and distant metastases.

Digestive System↗
PubMed1994

[Endocardial fibrosis in neuroendocrine tumors of the gastroenteropancreatic system].

Clinical characteristics, echocardiographic and Doppler echocardiographic findings, as well as serum levels of chromogranin A were recorded on 62 patients (27 women, 35 men; mean age 55 [11-83] years) with histologically confirmed tumours of the gastroenteropancreatic (GEP) system. Changes in the right heart were found in 14 patients (22%), club-like thickening of tricuspid leaflets in 13, tricuspid regurgitation in 14, stenosis in 2 and right atrial or right ventricular dilatation in 11 and 5, respectively. There was no difference between the patients with or without right-heart changes in regard to age, presence of carcinoid syndrome, duration of symptoms, primary tumour site, pattern of metastases, treatment or chromogranin A level. Two patients with nonfunctioning tumours had right-heart changes. While clinical and biochemical parameters did not identify patients with right-heart changes, echocardiography demonstrated all haemodynamically significant endocardial changes. Even patients with nonfunctioning GEP tumours should be regularly monitored by echocardiography.

Adolescent↗
PubMed1994

Sorting of synaptophysin into special vesicles in nonneuroendocrine epithelial cells.

Synaptophysin is a major transmembrane glycoprotein of a type of small vesicle with an electron-translucent content (SET vesicles), including the approximately 50-nm presynaptic vesicles in neuronal cells, and of similar, somewhat larger (< or = approximately 90 nm) vesicles (SLMV) in neuroendocrine (NE) cells. When certain epithelial non-NE cells, such as human hepatocellular carcinoma PLC cells, were cDNA transfected to synthesize synaptophysin, the new molecules appeared in specific SET vesicles. As this was in contrast to other reports that only NE cells were able to sort synaptophysin away from other plasma membrane proteins into presynaptic- or SLMV-type vesicles, we have further characterized the vesicles containing synaptophysin in transfected PLC cells. Using fractionation and immunoisolation techniques, we have separated different kinds of vesicles, and we have identified a distinct type of synaptophysin-rich, small (30-90-nm) vesicle that contains little, if any, protein of the constitutive secretory pathway marker hepatitis B surface antigen, of the fluid phase endocytosis marker HRP, and of the plasma membrane recycling endosomal marker transferrin receptor. In addition, we have found variously sized vesicles that contained both synaptophysin and transferrin receptor. A corresponding result was also obtained by direct visualization, using double-label immunofluorescence microscopy for the endocytotic markers and synaptophysin in confocal laser scan microscopy and in double-immunogold label electron microscopy. We conclude that diverse non-NE cells of epithelial nature are able to enrich the "foreign" molecule synaptophysin in a category of SET vesicles that are morphologically indistinguishable from SLMV of NE cells, including one type of vesicle in which synaptophysin is sorted away from endosomal marker proteins. Possible mechanisms of this sorting are discussed.

Biological Transport↗
PubMed1994

Localisation of neuroendocrine tumours of the upper gastrointestinal tract.

In order to localise neuroendocrine tumours of the foregut type (that is, of the stomach, duodenum, and pancreas), 18 patients were studied prospectively by endoscopic ultrasonography, computed tomography, transabdominal ultrasonography, magnetic resonance imaging, and somatostatin receptor scintigraphy. These 18 patients had a total of 25 primary tumour lesions which were verified histologically in tissue obtained by surgery or by ultrasound or endoscopy guided biopsy. Tumours were found in the stomach (n = 1), duodenum (n = 6), pancreas (n = 17), and liver (n = 1). Endoscopic ultrasonography had the highest sensitivity for tumour detection, followed by somatostatin receptor scintigraphy, computed tomography, transabdominal ultrasonography, and magnetic resonance imaging (88%, 52%, 36%, 32%, and 24% respectively). Endoscopic ultrasonography was especially sensitive in tumours smaller than 2 cm in diameter (88% v somatostatin receptor scintigraphy 35%; computed tomography 12%; transabdominal ultrasonography 6%; and magnetic resonance imaging 0%). Of 17 tumours located in the pancreas, endoscopic ultrasonography showed a sensitivity of 94% (somatostatin receptor scintigraphy 47%; computed tomography 47%; transabdominal ultrasonography 41%; and magnetic resonance imaging 29%). Of eight extrapancreatic tumours, six were identified by endoscopic ultrasonography, five by somatostatin receptor scintigraphy, and only one by computed tomography, transabdominal ultrasonography, and magnetic resonance imaging. One neuroendocrine tumour that was not detected by endoscopic ultrasonography was correctly identified by somatostatin receptor scintigraphy. Endoscopic ultrasound allowed correct determination of the tumour size and tumour spread into parapancreatic structures, especially the large vessels (T stage), in all 14 patients operated upon. The lymph node stage (N stage) was correctly determined in 10 of these 14 patients. In summary, endoscopic ultrasonography and somatostatin receptor scintigraphy were the most sensitive imaging methods for the localisation of these tumours and should be used as early diagnostic procedures to accurately stage neuroendocrine tumours of the foregut type.

Adolescent↗
PubMed1994

Detection of G-protein heterotrimers on large dense core and small synaptic vesicles of neuroendocrine and neuronal cells.

Heterotrimeric G proteins, initially believed to be exclusively present in the plasma membrane, have also been found to be associated with intracellular membrane compartments. There they are involved in various membrane trafficking processes including regulated secretion (reviewed in Bomsel, M., K. Mostov, Mol. Biol. Cell 3, 1317-1328 (1992)). Vesicles of two distinct types enter the regulated secretory pathway, i.e. large dense core vesicles and small synaptic vesicles, which differ in their membrane composition and content. Little is known about an association of heterotrimeric G proteins with regulated secretory vesicles, that would explain some aspects of the role heterotrimeric G proteins have during secretion. By immunofluorescence microscopy and immunoreplica analysis, we provide the first demonstration of the presence of complete sets of heterotrimeric G proteins, consisting of alpha-, beta-, and gamma-subunits, on large dense core vesicles from bovine adrenal medulla (chromaffin granules) and small synaptic vesicles from rodent and bovine brain. Each of the two types of secretory vesicles contains beta-subunits (at least beta 1 and beta 2), as well as gamma-subunits (at least gamma 2 or gamma 3). Interestingly, they differ in their composition of alpha-subunits. On small synaptic vesicles, we found two G(o) alpha-subunits (alpha o1 and alpha o2) and two Gi alpha-subunits (alpha i1 and alpha i2). In contrast, on chromaffin granules so far only one alpha o-subunit but no alpha i-subunits could be detected. Functional properties such as transmitter storage and/or exocytotic membrane fusion may be modulated by the various G-protein subunits associated with chromaffin granules and small synaptic vesicles.

Adrenal Medulla↗
PubMed1994

[Significance of false-positive findings of somatostatin receptor scintigraphy in diagnosis of neuroendocrine tumors of the gastroenteropancreatic system].

Localization of gastroenteropancreatic neuroendocrine tumors with conventional imaging techniques can be difficult. With the advent of the somatostatin-receptor scintigraphy the diagnosis of these could be improved. In this study 76 patients with immunohistologically proven neuroendocrine tumors were analysed by comparing somatostatin-receptor scintigraphy with conventional imaging techniques. Somatostatin-receptor-positive lesions were observed in 61 of all patients. Conventional imaging techniques (transabdominal ultrasonography, computerized tomographic scanning of the abdomen, magnetic resonance imaging of the abdomen) revealed neuroendocrine tumors in 42 patients (69%). A follow up of 19 patients with initially false positive somatostatin-receptor scintigrams showed that at least 6 patients had escaped the initial diagnosis by conventional techniques. All in all, our data support previous findings and demonstrate that somatostatin-receptor scintigraphy is a safe and sensitive procedure for in vivo imaging of gastroenteropancreatic neuroendocrine tumors. In addition, in certain cases, somatostatin-receptor scintigraphy is able to detect tumors that had escaped conventional imaging techniques.

Combined Modality Therapy↗
PubMed1993

Somatostatin-receptor imaging of neuroendocrine gastroenteropancreatic tumors.

BACKGROUND: Gastroenteropancreatic neuroendocrine tumors are often difficult to localize. This study was conducted to examine the value of somatostatin-receptor scintigraphy for visualization of gastroenteropancreatic neuroendocrine tumors. METHODS: Applying the recently developed indium-labeled somatostatin analogue 111In-pentetreotide to 40 patients with gastroenteropancreatic neuroendocrine tumors, the diagnostic power of pentreotide-receptor scintigraphy was evaluated in comparison with conventional imaging techniques. RESULTS: Expression of somatostatin receptors was observed in the majority of patients (11 of 17 in the foregut, 14 of 16 in the midgut, and 7 of 7 in metastatic neuroendocrine tumors with unknown primary). Comparative imaging by computerized tomography, magnetic resonance imaging, and transabdominal ultrasonography yielded false-negative results for somatostatin-receptor scintigraphy in 8 of 40 patients; however, in 16 patients, tumor tissue that had escaped conventional imaging techniques was detected by 111In-pentetreotide scintigraphy. CONCLUSIONS: 111In-pentetreotide scintigraphy is a practical, safe, and sensitive procedure for in vivo imaging of gastroenteropancreatic neuroendocrine tumors.

Adolescent↗
PubMed1993

Gastroenteropancreatic neuroendocrine tumours contain a common set of synaptic vesicle proteins and amino acid neurotransmitters.

Human neuroendocrine tumours of the gastroenteropancreatic system contain major integral membrane proteins of small synaptic vesicles of neurons, together with characteristic membrane polypeptides of large dense-core vesicles of neurons and neuroendocrine cells. The membrane polypeptides characteristic for small synaptic and large dense-core vesicles are detected in pheochromocytomas (n = 6), functional (n = 6) and non-functional (n = 6) foregut, and midgut carcinoids (n = 17). All gastroenteropancreatic tumours contain large amounts of amino acid neurotransmitters, i.e. glycine and glutamate. gamma-Aminobutyric acid, however, is only found in some foregut carcinoids. Thus, neuroendocrine gastroenteropancreatic tumours possess a vesicle type with a content and membrane composition similar to small synaptic vesicles of neurons.

Animals↗
PubMed1993

The small GTP-binding protein, Rab6p, is associated with both Golgi and post-Golgi synaptophysin-containing membranes during synaptogenesis of hypothalamic neurons in culture.

We have recently localized a small GTP-binding protein (Rab6p) thought to be involved in vesicular membrane transport, to the medial and trans-cisternae of the Golgi apparatus in NRK (normal rat kidney) cells. Here, we have localized and quantified Rab6p during the development in culture of embryonic neurons, up to synapse formation, and compared its subcellular distribution and level of expression to that of synaptophysin, a major integral membrane protein of small synaptic vesicles. Using immunocytochemistry (laser scanning confocal microscopy, immunoelectron microscopy), fractionation and immunoisolation methods, we show that during the early phase of synaptogenesis, Rab6p is associated with synaptophysin-containing membranes of a trans-Golgi subcompartment, post-Golgi vesicles and small synaptic vesicles or their precursors. Concomitantly, Rab6p undergoes translocation from cytosol to membranes and its level of expression increases. However, at late stages, the association of Rab6p to small synaptic vesicles sharply decreases and its level of expression plateaus. These findings suggest a role for Rab6p in the post-Golgi transport of synaptophysin, at an early step of the biogenesis of small synaptic vesicles.

Animals↗
PubMed1993

Pancreatic acinar cell carcinoma. An analysis of cell lineage markers, p53 expression, and Ki-ras mutation.

In a series of 22 pancreatic acinar cell carcinomas, including two acinar cystadenocarcinomas, cellular differentiation was analyzed by immunocytochemistry and electron microscopy. In addition, overexpression of p53 protein and Ki-ras codon 12 mutation was studied. Four of the 20 noncystic acinar cell carcinomas showed a pure acinar pattern, nine an acinar-solid, and seven a solid pattern. All tumors stained for at least one of the following pancreatic acinar markers: trypsin (21 of 22), lipase (19 of 22), chymotrypsin (13 of 22), phospholipase A2 (nine of 22), and pancreatic stone protein (19 of 22). One-third of the tumors expressed neuroendocrine markers (synaptophysin, eight of 22; chromogranin A, six of 21) and duct cell markers (CA19.9, nine of 21; B72.3, six of 21). Cellular coexpression of trypsin and synaptophysin was demonstrated in one tumor. Electron microscopy revealed zymogen granules (nine of nine). In only one of 16 tumors a Ki-ras mutation at codon 12 was found, whereas in none of 19 tumors could overexpression of p53 protein be demonstrated. The results suggest that acinar cell carcinomas show obvious capacity to differentiate into several directions, but nevertheless constitute an entity different from ductal adenocarcinomas or endocrine tumors.

Adolescent↗
PubMed1992

The amphicrine pancreatic cell line, AR42J, secretes GABA and amylase by separate regulated pathways.

Treatment of AR42J cells with dexamethasone leads to an enhanced formation of amylase-containing granules and facilitates their regulated secretion. Besides the exocrine properties, AR42J cells possess a specific uptake system for [3H]GABA. The stored GABA can be released upon potassium depolarisation in a Ca(2+)-dependent manner. After treatment with dexamethasone, potassium depolarisation fails to release GABA, but instead causes a Ca(2+)-dependent secretion of amylase. Since vesicles similar to small synaptic vesicles of neurons have been identified in AR42J cells, we suggest that the regulated GABA release is mediated by this vesicle type. It is tentatively speculated that other epithelial cells, which also contain small synaptic vesicles and amino acid neurotransmitters, may release them in a similar fashion.

Amylases↗
PubMed1992

Differential expression and subcellular localization of secretogranin II and synaptophysin during early development of mouse hypothalamic neurons in culture.

Mature neurons contain two distinct regulated secretory pathways, characterized electron microscopically by so-called large dense core vesicles and small synaptic vesicles, respectively. Each vesicle type is characterized by vesicle-specific proteins, such as the granins (chromogranins/secretogranins) for the matrix of large dense core vesicles and synaptophysin for the membrane of small synaptic vesicles. So far, no data exist on the biogenesis of these two distinct vesicle types during neuronal development. We have used secretogranin II and synaptophysin as markers for the biogenesis of these two vesicle types during the development of mouse hypothalamic neurons in culture, using immunocytochemistry and biochemical analyses. By immunofluorescence, we found that secretogranin II appears as early as synaptophysin, but in a subset of neurons only, and with different subcellular localizations. It was observed in cytoplasmic areas where little or no synaptophysin immunofluorescence was detected, such as lamellipodia, emerging neurites and growth cones. At later stages, the proportion of secretogranin II-containing varicosities remained steady whereas that of synaptophysin-containing varicosities increased dramatically. By quantitative analysis we found that the level of expression of synaptophysin increased several-fold during synaptogenesis whereas that of secretogranin II decreased. These data suggest that large dense core vesicles and small synaptic vesicles can be formed separately and expressed at different levels. They provide evidence for a differential biogenesis of these two distinct vesicle types.

Animals↗
PubMed1992

An amphicrine pancreatic cell line: AR42J cells combine exocrine and neuroendocrine properties.

The permanent cell culture line AR42J, derived from a rat pancreatic acinar carcinoma, is widely used for functional studies of exocrine pancreatic acinar cells. We now present evidence that these cells are amphicrine in that they contain zymogen granules as well as small (40-80 nm) neuroendocrine (NE) vesicles and typical neurotransmitters. Using the small NE vesicle-specific markers synaptophysin and "protein S.V.2", including synaptophysin cDNA probes, we have found that AR42J cells synthesize these proteins and contain vesicles harboring these proteins with biophysical properties similar to those of small NE vesicles. NE properties of these cells are further indicated by the presence of considerable amounts of stored amino acids (gamma-aminobutyric acid (GABA), glycine, glutamate) and by the presence of the GABA-synthesizing enzyme, glutamic acid decarboxylase. Finally, intermediate filament (IF) protein typing showed only cytokeratins 8 and 18, indicating that AR42J cells possess an IF protein complement indistinguishable from that of acinar and islet cells. Our results document the unusual case of a permanent cell line with combined exocrine and neuroendocrine properties that may be indicative of a derivation from a cell with multipotential character.

Animals↗
PubMed1992

Immunoluminometric assay of chromogranin A in serum with commercially available reagents.

Chromogranin A (Cg A) is a useful marker of neuroendocrine neoplasia in humans. Here we describe an immunoluminometric assay (ILMA) for measuring Cg A in serum, with use of a new tube as a solid phase. The new tube has a large surface area and is coated with a polyclonal antibody. Optimized coating conditions provide a high IgG adsorption to the polystyrene wall. Serum is added to the coated tube and incubated for 2 h or overnight. After washing the tubes, acridinium ester-labeled monoclonal antibody against Cg A is added and incubation is continued for 2 h. The tubes are washed again and the bound luminescence is measured. The assay is very sensitive (detection limit 1 microgram/L, if 25 microL of serum is used), is specific for human Cg A, and offers a wide dynamic range (1-1000 micrograms/L). The range in healthy humans is 10-53 micrograms/L (median 30 micrograms/L). The correlation of serum Cg A to creatinine and parathyroid hormone is described. We also report the influence of intravenous calcium injection on Cg A concentrations in serum.

Antibodies, Monoclonal↗