Cells Of Exocrine Pancreas Research Articles

Histology and ultrastructure of the hepatopancreas of the tigerfish, Hydrocynus forskahlii.

Study of the histology and ultrastructure of the hepatopancreas of the tigerfish, Hydrocynus forskahlii shows that the liver parenchyma is divided into irregularly shaped lobules, separated by the exocrine pancreas and associated connective tissue. The hepatocytes are arranged in interconnecting cords or laminae, two to three cell layers in thickness. Sinusoids separate the laminae. The spherical to oval-shaped hepatocytes contain large, round, centrally situated nuclei with prominent nucleoli. The cytoplasm of the hepatocytes contains abundant rough and smooth endoplasmic reticulae, free polysomes, and mitochondria. Exocrine pancreatic tissue is scattered throughout the liver. This tissue is encapsulated by an endothelium resting on a thin layer of connective tissue and is separated from the liver parenchyma by a sinusoid. The nuclei of the exocrine pancreas cells are spherical, basally situated within the cells, and contain dark nucleoli. Vesicular rough endoplasmic reticulae and secretory granules lie in the apical regions of the exocrine pancreas cells. © 1996 Wiley-Liss, Inc.

Histology and ultrastructure of the hepatopancreas of the tigerfish, Hydrocynus forskahlii

Study of the histology and ultrastructure of the hepatopancreas of the tigerfish, Hydrocynus forskahlii shows that the liver parenchyma is divided into irregularly shaped lobules, separated by the exocrine pancreas and associated connective tissue. The hepatocytes are arranged in interconnecting cords or laminae, two to three cell layers in thickness. Sinusoids separate the laminae. The spherical to oval-shaped hepatocytes contain large, round, centrally situated nuclei with prominent nucleoli. The cytoplasm of the hepatocytes contains abundant rough and smooth endoplasmic reticulae, free polysomes, and mitochondria. Exocrine pancreatic tissue is scattered throughout the liver. This tissue is encapsulated by an endothelium resting on a thin layer of connective tissue and is separated from the liver parenchyma by a sinusoid. The nuclei of the exocrine pancreas cells are spherical, basally situated within the cells, and contain dark nucleoli. Vesicular rough endoplasmic reticulae and secretory granules lie in the apical regions of the exocrine pancreas cells. © 1996 Wiley-Liss, Inc.

Detection of beta cell-specific DNA damage in streptozotocin-treated rats by in situ nick translation with immunostaining of alpha cells.

In situ nick translation with immunostaining of alpha cells could demonstrate the specific localization of streptozotocin- (STZ) induced DNA damage in beta cells using in vivo materials. The extent of DNA damage was determined through autoradiography by counting the number of grains in the nucleus of the alpha, beta, and exocrine cells of the rat pancreas. Subsequently, in situ nick translation with immunostaining of alpha cells was carried out in a pancreas pretreated with STZ. The number of grains observed in the beta cells of the STZ-treated groups was significantly higher than that in the control group. DNA damage of pancreatic beta cells due to STZ could be detected visually using in situ nick translation with immunostaining of alpha cells. Moreover, it was also possible to compare the DNA damage in the individual cells of the pancreas.

Structure of the exocrine pancreas of flounder (Paralichthys olivaceus): immunological localization of zymogen granules in the digestive tract using anti‐trypsinogen antibody

Most fish have a diffuse exocrine pancreas located around the internal organs, but its exact structure is not well understood. In this study, we developed a method for analysing zymogen granules using anti‐fish trypsinogen, and the structure of the flounder pancreas was examined by localizing pancreatic exocrine cells. Anionic trypsinogen was purified from eel pancreas by chromatography using DE‐52‐cellulose and Superose 12 columns. Rabbit antiserum (anti‐eel trypsinogen antibody: anti‐eTrg) was raised against purified trypsinogen. Immunoblotting analysis demonstrated that anti‐eTrg recognized two isoforms of eel trypsinogens and trypsins. This antibody also reacted with flounder trypsin. Zymogen granules containing trypsinogen were visualized within pancreatic glandular cells of the flounder Paralichthys olivaceus by immunohistochemistry using anti‐eTrg. The flounder exocrine pancreas was reconstructed from observations of serial sections of digestive tract. The exocrine pancreas cells were localized along the hepatic portal vein and the veins running to the portal vein from the intestine, stomach, pyloric appendage and spleen, forming a highly branched tissue. Interlobular ducts from pancreatic acini formed a pancreatic duct covered with smooth muscle at the point where it met (but did not join) the bile duct. The pancreatic duct opened into the intestinal lumen at the basal portion of the third pyloric appendage in parallel with the bile duct.

Neuronal and endothelial nitric oxide synthase immunoreactivity and NADPH-diaphorase staining in rat and human pancreas: influence of fixation

In this study, we wished to clarify the distribution and co-localization of nitric oxide synthase and NA-DPH-diaphorase (NADPH-d) in nerve cells, nerve fibres and parenchymal cells in exocrine and endocrine pancreas, and to assess the influence of fixation on the staining pattern obtained. For this purpose, we applied nitric oxide synthase immunocytochemistry and NADPH-d histochemistry to rat and human pancreas under different fixation conditions. Antibodies to neuronal and endothelial nitric oxide synthase were similarly applied. We found complete co-localization of neuronal nitric oxide synthase and NADPH-d in ganglion cells, and in nerve fibres around acini, excretory ducts, blood vessels and in islets of Langerhans of rat and human pancreas. Immunoreactivity for endothelial nitric oxide synthase was co-localized with NADPH-d in endothelial cells. However, in NADPH-d reactive islet and ductal epithelial cells we could detect neither brain nor endothelial nitric oxide synthase immunoreactivity with any fixation protocol applied. There were marked differences in NADPH-d staining of both neurons and parenchymal cells under different fixation conditions. These results indicate the existence of different types of NADPH-d, which are associated or not associated with nitric oxide synthase(s), and which are differently influenced by various fixation procedures in rat and human pancreas.

Glucocorticoid receptor binding site in the mouse alpha-amylase 2 gene mediates response to the hormone.

Amylase gene expression has been shown to be positively regulated by glucocorticoids. Previous reports have suggested that this effect is indirect. We have addressed this question in a mouse exocrine pancreas cell line, 266-6, in which basal level of expression of amylase mRNA is low but inducible by glucocorticoids. In these cells the effect of glucocorticoids is not inhibited by cycloheximide at early time points. Reporter plasmids containing 224 base pairs of mouse amylase 5'-flanking DNA are positively regulated by glucocorticoids in gene transfer experiments. Glucocorticoid receptor purified from rat liver binds to the amylase promoter from position -56 to -33 and at the start of transcription. Site-directed mutation at the upstream position (-47 to -42) eliminates response to glucocorticoids in transient gene transfer experiments. Thus, glucocorticoid regulation of the mouse amylase gene is a direct effect and is mediated via a receptor binding site in the promoter region of the gene. Inhibition of the hormone response by cycloheximide at later time points after induction suggests the additional requirement for a short-lived factor. The DNA binding domain of the glucocorticoid receptor binds to a single site in the amylase promoter as a monomer, suggesting that both receptor binding sites as well as an additional short-lived factor are required to obtain induction.

Relationships among residual β cells, exocrine pancreas, and islet cell antibodies in insulin-dependent diabetes mellitus

Quantitative analysis was performed using computerized morphometry on the relationships between residual β cells, pancreatic exocrine glands, and islet cell antibodies (ICA) in 14 pancreata of insulin-dependent diabetic (IDDM) patients. Both pancreatic exocrine glands and β cells were markedly reduced in weight in IDDM patients compared with non-insulin-dependent diabetic (NIDDM) patients or nondiabetic controls. β cells were preserved in six cases and were completely abolished in eight cases. In nine IDDM pancreata weighed at autopsy, the weights of pancreatic exocrine glands in six patients with either no or virtually no residual β cells (32.3 ± 1.6 g) were greater than those in three patients with residual β cells (23.1 ± 2.5 g, P < .05). Infiltration of lymphocytes positive for leukocyte common antigen (LCA) around the islet was observed in only one case with ICA and residual β cells. Infiltration of LCA-positive lymphocytes around pancreatic acinar cells was observed in 50% (six of 12) of patients examined. The weight of pancreatic exocrine glands in patients with LCA-positive lymphocyte infiltration (26.2 ± 2.3 g) was lower than that in patients without this condition (33.2 ± 2.4 g, P < .05). Pancreatic cytokeratin autoantibodies (PKA) were detected in four of 10 patients examined. In addition, all four ICA-positive patients had residual β cells, while only one of seven ICA-negative patients had residual β cells ( P = .03). The better preservation of exocrine pancreas glands in patients without residual β cells than in those with residual β cells cannot be explained by the hypothesis that atrophy of the pancreatic exocrine gland is due to loss of the trophic action of insulin on acinar cells. Exocrine pancreatic atrophy accompanied by infiltration of LCA-positive lymphocytes around the acinar cells suggests primary involvement of the exocrine pancreas in the autoimmune pathological process of IDDM. The contribution of residual β cells to the generation of ICA is also suggested.

Identification and localization of 5-HT1P receptors in the guinea pig pancreas.

Enteropancreatic 5-hydroxytryptamine (5-HT)-immunoreactive nerve fibers have recently been described. To test the hypothesis that pancreas contains 5-HT receptors similar to those that characterize the enteric nervous system, binding of 5-[3H]HT to pancreatic membranes was investigated. Membranes were prepared by differential centrifugation from guinea pig pancreas, and binding of 5-[3H]HT was measured by use of a rapid filtration assay. Alternatively, 5-[3H]HT-binding sites were analyzed and located by radioautography on frozen sections of unfixed pancreas. The 5-HT1P receptor-specific antagonist, N-acetyl-5-hydroxytryptophyl-5-hydroxytryptophan amide (5-HTP-DP), was used to define nonspecific binding in all assays. High-affinity reversible binding of 5-[3H]HT to pancreatic membranes was found. Good agreement was observed when Kd was determined from saturation isotherms (Kd = 2.1 +/- 0.4 nM; Bmax = 0.6 +/- 0.07 pmol/mg protein) or from the ratio of association and dissociation constants (2.7 +/- 0.04 nM). As is true of 5-[3H]HT binding to enteric 5-HT1P receptors, binding of 5-[3H]HT to pancreatic membranes was inhibited by 5-HTP-DP, 5- and 6-hydroxyindalpine (agonists at 5-HT1P receptors), and guanine nucleotides. In contrast, binding of 5-[3H]HT to pancreatic membranes was not inhibited by a variety of compounds known to bind to other types of 5-HT receptor, to the neuronal 5-HT transporter, to alpha- or beta-adrenoceptors, or to dopamine receptors. We concluded that pancreas contains 5-HT1P receptors with properties similar to those of bowel. Radioautography and immunocytochemistry with an anti-idiotypic antibody that recognizes 5-HT receptors revealed that pancreatic 5-HT1P receptors (identified with appropriate inhibitors of 5-[3H]HT or antibody binding) were concentrated in ganglia, nerves, and unidentified cells in both exocrine and endocrine pancreas. These observations suggest that future studies of 5-HT's role in pancreatic physiology should concentrate on enteropancreatic innervation and 5-HT1P receptors.

Die histologie en ultrastruktuur van die hepatopankreas van die bloukurper &lt;i&gt;Oreochromis mossambicus&lt;i/&gt;

The histology and ultrastructure of the hepatopancreas of the Mozambique tilapia O. mossambicus are described. The liver is surrounded by a thin connective tissue capsule. The hepatocyte arrangement shows as lobules, with the hepatocyte cords that radiate and anastomose from a central vein. Borders of individual liver lobules do not show clearly, as do the few triads found in the liver of O. mossambicus. Each hepatocyte contains a single round nucleus with a prominent nucleolus. The rough endoplasmic reticulum appears in two or more rows around the nuclei and in close proximity to the plastnalemma of the hepatocytes. The remaining cytoplasmic organelles are scattered throughout the hepatocyte cytoplasm. The exocrine pancreas cells are centred around the portal veins. The nuclei of these cells are spherical and hasally situated in the cubiform to cylindriform cells. Well developed rough endoplasmic reticula - vesicular, tubular and circular in appearance - as well as secretory granules, apically situated in the cells, are present.

Morphometry and Elemental Analysis of Rat Exocrine Pancreas following Administration of Trypsin Inhibitor

The morphological responses of the exocrine pancreas of the adult male rat to soybean trypsin inhibitor (STI) were studied by ultrastructural morphometry and electron probe X-ray microanalysis. STI administered orally in drinking water for 14 days resulted in a 72% increase in the wet weight of the pancreas. This enlargement was due, largely, to an increase in acinar cell mass. Volume increases in the acinar cell mass and extra-acinar cell compartment were 72 and 30%, respectively. The estimated total number of acinar cells in the mean exocrine pancreas was 500 million in the control and 630 million in the experimental group, representing an increase of 27%. Acinar cell volume was 1,790 microns 3 for the control and 2,457 microns 3 for the STI group. The pronounced morphometric changes of the organelles in the STI group were: the mean nucleolar volume increased by 56%; the volume of zymogen granular mass per cell increased by 93%; the volume of the Golgi complex and the condensing vacuoles per cell increased by 52 and 100%, respectively, whereas the membrane area of the Golgi complex and the condensing vacuoles increased by 98 and 47%, respectively. Spectral analysis of seven elements (Na, Mg, P, S, Cl, K and Ca) showed significant changes for nuclei, zymogen granules and mitochondria following STI: nuclei showed Na, P, K increased; zymogen granules showed Na, P, S, K increased, Cl decreased; mitochondrial particles showed Mg, P, Cl, Ca increased, and the mitochondrial matrix showed S decreased. The persistent uptake of STI probably resulted in a continual release of a trophic hormone acting on pancreatic tissue components, consequently causing hyperplasia and hypertrophy of the exocrine pancreas to accommodate a heightened demand for synthesis of exportable proteins.

Glucocorticoids effects on exocrine pancreatic secretion in caeruleininduced acute pancreatitis in the rat

The present work reports on exocrine pancreatic secretion in control rats, adrenalectomized rats and hydrocortisone-treated (10 mg/Kg/d) rats during 7 days, under normal conditions and after induction of acute pancreatitis with caerulein (20 micrograms/Kg) by 4 subcutaneous injections at hourly intervals. Pancreatic secretion was seen to be affected by the procedure of adrenalectomy, which led to a marked reduction in the secretion of proteins and amylase with respect to control values. This was probably due to the decrease occurring in the zymogen granules in the acinar cells of the exocrine pancreas, a phenomenon which also led to a decrease in pancreatic weight observed in these animals. Treatment with hydrocortisone induced a decrease in the secretion of proteins and amylase, as well as an increase in pancreatic weight. This agrees with the accepted hypothesis that large amounts glucocorticoids stimulate the synthesis and storage of proteins in the exocrine pancreas, reducing the secretory phase. The administration of high doses of caerulein under these conditions led to acute pancreatitis in the three groups of animals. This was paralleled by a dramatic decrease in protein and amylase secretion and by severe interstitial edema of the pancreas and by increases in serum amylase values. In the case of the animals treated previously with hydrocortisone, the latter were tripled with respect to the control animals. The conclusion is offered that since the storage of enzyme proteins is governed by glucocorticoids, which furthermore increase the sensitivity of the acinar cells to stimulation by secretagogues, the administration of these substances during the development of pancreatic lesions such as acute pancreatitis is highly compromising to the organism.

Tissue distribution of [formula omitted] p25A, a [formula omitted] p21-like GTP-binding protein, studied by use of a specific monoclonal antibody

We made a monoclonal antibody specifically recognizing smg p25A among many ras p21-like GTP-binding proteins and investigated the tissue distribution of smg p25A by use of this antibody. By immunoblot analysis, smg p25A was detected in rat brain and bovine adrenal medulla but not in bovine adrenal cortex or other rat tissues including thymus, spleen, lung, heart, liver and kidney. However, by immunocytochemical studies, smg p25A was detected not only in the synaptic areas of rat brain and the chromaffin cells of bovine adrenal medulla but also in the endocrine cells of rat pancreatic islets, the acinar cells of rat exocrine pancreas and the exocrine cells of rat submaxillary gland. These results suggest that smg p25A is involved in the regulation of secretory processes not only in synapses but also in other endocrine and exocrine secretory cells.

Characterization of MgATP-Driven H+ uptake in to a microsomal vesicle franction from rat pancreatic acinar cells

In microsomal vesicles, as isolated from exocrine pancreas cells, MgATP-driven H+ transport was evaluated by measuring H+-dependent accumulation of acridine orange (AO). Active H+ uptake showed an absolute requirement for ATP with simple Michaelis-Menten kinetics (Km for ATP 0.43 mmol/liter) with a Hill coefficient of 0.99. H+ transport was maximal at an external pH of 6.7, generating an intravesicular pH of 4.8. MgATP-dependent H+ accumulation was abolished by protonophores, such as nigericin (10(-6) mol/liter) or CCCP (10(-5) mol/liter), and by inhibitors of nonmitochondrial H+ ATPases, such as NEM or NBD-Cl, at a concentration of 10(-5) mol/liter. Inhibitors of both mitochondrial and nonmitochondrial H+ pumps, such as DCCD (10(-5) mol/liter) or Dio 9 (0.25 mg/ml), reduced microsomal H+ transport by about 90%. Vanadate (2 x 10(-3) mol/liter), a blocker of those ATPases, which form a phosphorylated intermediate, did not inhibit H+ transport. The stilbene derivative DIDS (10(-4) mol/liter), which inhibits anion transport systems, abolished H+ transport completely. MgATP-dependent H+ transport was found to be anion dependent in the sequence Cl- greater than Br- greater than gluconate-; in the presence of SO2-4, CH3COO- or No-3, no H+ transport was observed. MgATP-dependent H+ accumulation was also cation dependent in the sequence K+ greater than Li+ greater than Na+ = choline+. As shown by dissipation experiments in the presence of different ion gradients and ionophores, both a Cl- and a K+ conductance, as well as a small H+ conductance, were found in the microsomal membranes. When membranes containing the H+ pump were further purified by Percoll gradient centrifugation (ninefold enrichment compared to homogenate), no correlation with markers for endoplasmic reticulum, mitochondria, plasma membranes, zymogen granules or Golgi membranes was found. The present data indicate that the H+ pump located in microsomes from rat exocrine pancreas is a vacuolar- or "V" -type H+ ATPase and has most similarities to that described in endoplasmic reticulum, Golgi apparatus or endosomes.

The nonselective cation channel in the basolateral membrane of rat exocrine pancreas

Nonselective Ca2+-sensitive cation channels in the basolateral membrane of isolated cells of the rat exocrine pancreas were investigated with the patch clamp technique. With 1.3 mmol/l Ca2+ on the cytosolic side, the mean open-state probability Po of one channel was about 0.5. In inside-out oriented cell-excised membrane patches the substances diphenylamine-2-carboxylic acid (DPC), 5-nitro-2-(3-phenelpropylamino)-benzoic acid (NPPB) and 3',5-dichlorodiphenylamine-2-carboxylic acid (DCDPC) were applied to the cytosolic side. These compounds inhibited the nonselective cation channels by increasing the mean channel closed time (slow block). 100 mumol/l of NPPB or DPC decreased Po from 0.5 (control conditions) to 0.2 and 0.04, respectively, whereas 100 mumol/l of DCDPC blocked the channel completely. All effects were reversible. 1 mmol/l quinine also reduced Po, but in contrast to the above mentioned substances, it induced fast flickering. Ba2+ (70 mmol/l) and tetraethylammonium (TEA+; 20 mmol/l) had no effects. We investigated also the stilbene disulfonates 4-acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic acid (SITS), 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS) and 4,4'-dinitro-2,2'-stilbenedisulfonate (DNDS). 10 mumol/l SITS applied to the cytosolic side increased Po from 0.5 to 0.7 and with 100 mumol/l SITS the channels remained nearly permanently in its open state (Po approximately equal to 1). A similar activation of the channels was also observed with DIDS and DNDS. These effects were poorly reversible. The stilbene disulfonates acted by increasing the channel mean open time. When the channel was inactivated by decreasing bath Ca2+ concentration to 0.1 mumol/l, addition of 100 mumol/l of SITS had no effect. Similarly, reducing bath Ca2+ concentration from 1.3 mmol/l in presence of 100 mumol/l SITS (channels are maximally activated) to 0.1 mumol/l, inactivated the channels completely. These results demonstrate, that SITS can only activate the channels in the presence of Ca2+. SITS had no effects, when applied to the extracellular side in out-side out patches.(ABSTRACT TRUNCATED AT 400 WORDS)

1] Membrane transport of calcium: An overview

Publisher Summary This chapter focuses on the membrane transport of calcium. Ca 2+ signaling plays an important role in numerous areas of cell biochemistry and physiology, ranging from exocrine and hormonal secretion, to muscle and nonmuscle motility, to the activity and regulation of several important metabolic pathways. The central element in the signaling function of Ca 2+ is its reversible complexation by specific proteins, which bind Ca 2+ in micromolar or submicromolar concentrations with optimal affinity in the presence of much larger concentrations of other divalent and monovalent cations, such as Mg 2+ , Na + , and K + . Many of these proteins are dissolved in the cytosol or organized in nonmembrane structures, such as the myofibrils. Interest in endoplasmic reticulum as a Ca 2+ reservoir has been heightened by the finding that inositol trisphosphate, one of the two important catabolites of phosphatidylinositol diphosphate, specifically releases Ca 2+ from endoplasmic reticulum in response to a number of plasma membrane stimuli that activate a phospholipase. The release was originally demonstrated in permeabilized exocrine pancreas cells, but more recently, has been shown in isolated endoplasmic reticulum vesicles. In the latter system, however, particular conditions must be applied.

Quantitative energy dispersive X-ray microanalysis of eight elements in pancreatic endocrine and exocrine cells after cryo-fixation

Quantitative X-ray microanalysis of 8 elements was performed on ultrathin, freeze-dried sections of islets and pancreas pieces from non-inbred ob/ob-mice. Diffusion of elements was reduced to a minimum by rapidly freezing the tissue samples between nitrogen-cooled polished copper surfaces and avoiding the use of chemical fixatives and stains. The ultrastructural morphology was adequately maintained to allow measurements on secretory granules, mitochondria, cell nuclei, and cytoplasm free of these organelles. The distribution of the various elements between cellular compartments was similar in islet beta-cells and exocrine pancreas cells. However, the insulin secretory granules were outstanding in exhibiting the highest concentrations of zinc and calcium. In comparison with cytoplasm in the beta-cells, the insulin granules accumulated calcium 2-fold and zinc as much as 40-fold. As no correlation could be made for endoplasmic reticulum in the cytoplasmic measurements areas, the true accumulations above cytosol are likely to be even higher.

Selective regulation of trypsin gene expression by calcium and by glucose starvation in a rat exocrine pancreas cell line.

Treatment of the rat pancreatic acinar cell line AR4-2J with the calcium ionophore A23187 selectively increases, within a few hours, the steady-state level of trypsin mRNA. Addition of the tumor-promoting phorbol ester phorbol 12-myristate 13-acetate potentiates the calcium-induced increase. The mRNA level of the other tested exocrine pancreatic genes decreases. These results were confirmed by DNA transfection experiments, using the 5' flanking region of the trypsin and chymotrypsin genes linked to the coding sequence of the chloramphenicol acetyltransferase (CAT) gene. In calcium-induced cells transfected with the trypsin constructs, an increase in CAT activity was observed, whereas the chymotrypsin constructs revealed a decreased CAT activity. Glucose starvation of AR4-2J cells similarly elicited a selective increase in trypsin mRNA. This selective regulation of trypsin may reflect its role as the key activator of the other zymogen species.

Two Different Factors Act Separately or Together To Specify Functionally Distinct Activities at a Single Transcriptional Enhancer

The expression of genes fused downstream of the Moloney murine sarcoma virus (MoMSV) long terminal repeat is stimulated by glucocorticoids. We mapped the glucocorticoid response element that conferred this hormonal regulation and found that it is a hormone-dependent transcriptional enhancer, designated Sg; it resides within DNA fragments that also carry a previously described enhancer element (B. Levinson, G. Khoury, G. Vande Woude, and P. Gruss, Nature [London] 295:568-572, 1982), here termed Sa, whose activity is independent of the hormone. Nuclease footprinting revealed that purified glucocorticoid receptor bound at multiple discrete sites within and at the borders of the tandemly repeated sequence motif that defines Sa. The Sa and Sg activities stimulated the apparent efficiency of cognate or heterologous promoter utilization, individually providing modest enhancement and in concert yielding higher levels of activity. A deletion mutant lacking most of the tandem repeat but retaining a single receptor footprint sequence lost Sa activity but still conferred Sg activity. The two enhancer components could also be distinguished physiologically: both were operative within cultured rat fibroblasts, but only Sg activity was detectable in rat exocrine pancreas cells. Therefore, the sequence determinants of Sa and Sg activity may be interdigitated, and when both components are active, the receptor and a putative Sa factor can apparently bind and act simultaneously. We concluded that MoMSV enhancer activity is effected by at least two distinct binding factors, suggesting that combinatorial regulation of promoter function can be mediated even from a single genetic element.

Two different factors act separately or together to specify functionally distinct activities at a single transcriptional enhancer.

The expression of genes fused downstream of the Moloney murine sarcoma virus (MoMSV) long terminal repeat is stimulated by glucocorticoids. We mapped the glucocorticoid response element that conferred this hormonal regulation and found that it is a hormone-dependent transcriptional enhancer, designated Sg; it resides within DNA fragments that also carry a previously described enhancer element (B. Levinson, G. Khoury, G. Vande Woude, and P. Gruss, Nature [London] 295:568-572, 1982), here termed Sa, whose activity is independent of the hormone. Nuclease footprinting revealed that purified glucocorticoid receptor bound at multiple discrete sites within and at the borders of the tandemly repeated sequence motif that defines Sa. The Sa and Sg activities stimulated the apparent efficiency of cognate or heterologous promoter utilization, individually providing modest enhancement and in concert yielding higher levels of activity. A deletion mutant lacking most of the tandem repeat but retaining a single receptor footprint sequence lost Sa activity but still conferred Sg activity. The two enhancer components could also be distinguished physiologically: both were operative within cultured rat fibroblasts, but only Sg activity was detectable in rat exocrine pancreas cells. Therefore, the sequence determinants of Sa and Sg activity may be interdigitated, and when both components are active, the receptor and a putative Sa factor can apparently bind and act simultaneously. We concluded that MoMSV enhancer activity is effected by at least two distinct binding factors, suggesting that combinatorial regulation of promoter function can be mediated even from a single genetic element.

A quantitative immuno-electronmicroscopic study of amylase and chymotrypsinogen in peri- and tele-insular cells of the rat exocrine pancreas.

Malaisse-Lagae demonstrated in 1975 that peri-insular (PI) cells and tele-insular (TI) cells produce amylase (Am) and chymotrypsinogen (Ch) in a different ratio. These biochemical measurements are in contradiction with recent observations of Bendayan (1985), who found that the Am/Ch ratio measured with the protein A-gold technique applied to ultrathin Epon sections was the same in PI and TI cells. We have previously shown (Posthuma et al., 1984) that experimentally induced changes in Am and Ch content of rat pancreas are quantitatively reflected by immuno-gold labeling of zymogen granules in cryosections. Here we applied the same technique to compare the Am/Ch labeling density ratios in PI and TI pancreatic cells. To ascertain constancy of experimental conditions, we used ultrathin cryosections from tissue blocks consisting of TI and PI tissue elements. Consecutive sections of these blocks were alternatively immunolabeled for Am and Ch, using protein A-gold as marker. The density of gold particles over zymogen granules of both PI and TI cells was measured. It appeared that the Am/Ch labeling density ratio was significantly lower in PI than in TI cells. This difference resulted from a lower Am labeling as well as higher Ch labeling density over zymogen granules in PI cells.