Glucokinase in B-cell-depleted islets of Langerhans

Glucose phosphorylation was studied in B-cell-enriched or in B-cell-depleted pancreatic islets from normal or streptozotocin-diabetic rats, respectively, using quantitative histochemical procedures. The data indicate that B-cell-enriched preparations from normal animals and whole islets from normals, diabetics, and insulin-treated diabetic animals have comparable glucokinase activities. Average maximum velocities were (mmol/kg dry tissue/hr) 134.1 +/- 7.3 for whole islets and 125.6 +/- 10.7 for the B-cell-enriched preparations from normal rats, 143.1 +/- 13.6 for B-cell-depleted islets from diabetic rats, and 124.4 +/- 10.7 for B-cell-depleted islets from insulin-treated diabetic animals. The Kmax for glucose of the enzyme in islets from untreated diabetic rats was 16 mM, comparable to the Kmax found for glucokinase from normal rat islets. Mannoheptulose, previously shown to be a competitive inhibitor of glucokinase from liver and normal islets, also inhibited glucokinase in B-cell-depleted islets from diabetic rats. The data indicate that glucokinase is not selectively located in the B-cell, as was previously assumed, but is also found in A- and/or D-cells of diabetic rats. This observation raises significant questions about the functional role of islet glucokinase under control and diabetic conditions.     

Increased filamentous actin in islets of Langerhans from fasted hamsters

We measured pools of soluble and sedimentable actin in hamster islets using a new DNase I binding/immunoprecipitation assay. Islets from fed and fasted hamsters contained the same amount of actin when expressed per microgram of protein. In three experiments the sedimentable (filamentous) actin increased significantly in islets from fasted hamsters. Recovery determinations demonstrate that during fasting the sedimentable actin pool (F) is increased as the soluble (G) actin pool decreases. These results suggest that the microfilament system is affected by the metabolic state of hamsters and may be responsible, in part, for inhibiting insulin secretion during fasting.     

Organ culture of the islets of Langerhans from young and senescent rats

Summary The B-cells of the endocrine pancreas constitute an adequate model for in vitro study of the aging process in highly differentiated cells. In the present study, collagenase-isolated islets of Langerhans from young and senescent rats were cultured up to 28 days. The response of the B-cells to the stimulatory conditions of the culture medium involved the nucleus, ribosomes, endoplasmic reticulum, Golgi apparatus, and secretory granules. Correlated data from light microscopy, electron microscopy, and insulin radioimmunoassay show that the differentiation and function of senescent B-cells are maintained in culture, as it has been proven for the B-cells of younger animals. On the other hand, signs of cytological deficiency not directly concerned with the specific function of B-cells were observed: abnormal mitochondria and lysosomes are more numerous in the senescent B-cells. The proliferative capacity of the B-cells of aged rats is reduced. Send offprint requests to: Université Catholique de Louvain, Unité de Morphologie animale, Place Croix du Sud, 5, B 1348 Louvain-la-Neuve, BelgiumM.C.M. is currently associated with the Unité d'Histologie, Université de Louvain     

Adventures with insulin in the islets of Langerhans

Insulin is a small but beautifully organized protein with a unique two-chain structure, the first protein to be sequenced. The mechanism of its biosynthesis invited much initial speculation but was finally clarified by the discovery of proinsulin, its single-chain precursor. The rich present-day field of protein precursor processing via post-translational proteolysis within the secretory pathway arose in the early 1970s as an offshoot of studies on insulin biosynthesis, which provided a novel paradigm for the generation of many other small neuroendocrine peptides. Before long, this mechanism was also found to play a role in the production of a much wider spectrum of proteins traversing the secretory pathway (receptors, growth factors, blood-clotting components, and even many viral envelope proteins) occurring in almost all eukaryotic cells. Indeed, yeast provided a key clue in the search for the proprotein convertases, the endoproteases that work along with carboxypeptidases and other modifying enzymes, such as the amidating enzyme complex (PAM), in converting inactive or less active precursor proteins into their fully active peptide products. In this "Reflections" article, I have tried to recount the people and events in my life that led to my involvement first in basic biochemical research and then on to insulin, proinsulin, and many relevant related areas that continue to fascinate and challenge my colleagues and me, as well as many other biomedical scientists today, as diabetes mellitus increasingly threatens human health throughout our contemporary world.     

Nestin-expressing cells in the pancreatic islets of Langerhans

The pancreatic islets of Langerhans produce several peptide hormones, predominantly the metabolically active hormones insulin and glucagon, which are critical for maintaining normal fuel homeostasis. Some evidence exists that pancreatic endocrine cells turn over at a slow rate and can regenerate in certain conditions. This could be due to the presence of pluripotent cells residing in the pancreas. Recently the intermediate filament protein nestin has been identified to be a marker for a multipotent stem cell in the central nervous system. Given the similarity between the pancreatic islets and neuronal cells, we hypothesized that stem cells expressing nestin might be present in the pancreas. Here we present evidence that a subset of cells in the pancreatic islets express the stem cell marker nestin. These cells might serve as precursors of differentiated pancreatic endocrine cells.     

Phosphorylation reactions in islets of Langerhans.

The possible significance of phosphorylation reaction in islets of Langerhans in relation to the secretion of insulin is discussed. The secretagogues, glucose and its metabolites, cAMP and Ca++ and their influences on protein-kinase activity are given particular attention. The data obtained by the authors, as well as by other groups, are in agreement that cAMP is a potent stimulator of protein kinase activity. Glucose and its metabolites influenced protein kinase activity in one instance. Ca++ in supraphysiological amounts inhibited protein phosphorylation. The links between phosphorylation reactions and insulin secretion are, at the present time, conjectural.     

Estrogen receptors in the islets of Langerhans of baboons

Previous clinical studies have indicated that during pregnancy and following administration of contraceptives women show altered carbohydrate metabolism. We performed autoradiographic studies using 3H-estradiol-17 beta and 3H-dihydrotestosterone on male and female baboons. Discrete sites of localization of exposed photographic emulsion were observed over nuclei of cells of the islets of Langerhans of the pancreases of baboons injected with estrogen but not over those of baboons injected with androgen. These observations that islet cells contain specific receptors for estrogen when combined with the clinical observations, suggest that estrogens have a direct effect on the islet cells that may modulate the release of insulin.     

Regeneration of the islets of Langerhans in the guinea pig

Summary The regeneration of the islets of Langerhans in the guinea pig was studied after intravenous injection of alloxan. A number of cells in the islets was destroyed within 24 h after alloxan, but after 48 h there was a rapid proliferation of the surviving cells of the islets. This depended on the dosage of the drug as well as the timing. Electron microscopy of the islet at 48 h showed that the dividing cells had small electron dense granules and resembled a subtype of normal A cells, whose function is not yet known. There were also many agranular cells in the islet. These two groups of cells seen in the regenerating islet could be precursor cells, which could differentiate into cells. There was no evidence for transformation of duct cells or acinar cells into islet cells. None of the guinea pigs became permanently diabetic. This was probably due to inadequate dosage which resulted in only partial degeneration of the cells followed by regeneration and recovery. There was also some regeneration of the liver, kidney and the adrenal cortex following alloxan.The author is grateful to Professor R. Barer for his guidance and for providing the facilities for this study. Thanks are also due to Mrs. D. Barraclough for technical assistance and to Mrs. M. Hollingsworth for assistance with the photographsThis work was financed by a grant from the Cystic Fibrosis Research Trust     

Glucose diffusion in pancreatic islets of Langerhans.

We investigate the time required for glucose to diffuse through an isolated pancreatic islet of Langerhans and reach an equilibrium. This question is relevant in the context of in vitro electrophysiological studies of the response of an islet to step changes in the bath glucose concentration. Islet cells are electrically coupled by gap junctions, so nonuniformities in islet glucose concentration may be reflected in the activity of cells on the islet periphery, where electrical recordings are made. Using a mathematical model of hindered glucose diffusion, we investigate the effects of the islet porosity and the permeability of a surrounding layer of acinar cells. A major factor in the determination of the equilibrium time is the transport of glucose into islet beta-cells, which removes glucose from the interstitial spaces where diffusion occurs. This transport is incorporated by using a model of the GLUT-2 glucose transporter. We find that several minutes are required for the islet to equilibrate to a 10 mM change in bath glucose, a typical protocol in islet experiments. It is therefore likely that in electrophysiological islet experiments the glucose distribution is nonuniform for several minutes after a step change in bath glucose. The delay in glucose penetration to the inner portions of the islet may be a major contributing factor to the 1-2-min delay in islet electrical activity typically observed after bath application of a stimulatory concentration of glucose.