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Methods have been developed for the isolation on a semi-micro scale of a plasma membrane-enriched fraction from rat islets of Langerhans. An important feature of these experiments is the use of 125I-labeled wheat germ agglutinin as a specific probe for plasma membrane-containing fractions. The partly purified plasma membrane fraction had a density in sucrose of about 1.10 and was enriched in the a
The overall dynamics of glucose-induced insulin release was strikingly similar in dispersed cells and intact islets perifused in parallel. Both preparations exhibited a latency of 1-2 min, after which period there was a brisk rise of insulin release followed by a sustained second phase. During the second phase, insulin release from dispersed cells attained a stable plateau rate, whereas the releas
Pancreatic islets rich in beta-cells were isolated from non-inbred ob/ob-mice and used for studying various aspects of the function of the plasma membrane. A review is given of the authors' work along the following lines: the role of transmembrane transport or membrane binding in the recognition of insulin-releasing sugars, amino acids, sulfonylureas, and sulphydryl-blocking agents; the role of cy
The effects of thiol compounds on insulin release were studied in microdissected pancreatic islets of non-inbred ob/ob mice. In control experiments the reactivity of thiols against 6,6′-dithiodinicotinic acid and the degradation of mouse insulin were measured. At a concentration of 0.1 mM, 1-thio-D-glucose or reduced glutathione potentiated the insulin-releasing action of 10 mM D-glucose without a
Six previously pancreatectomized dogs were transplanted with ductligated, pancreatic allografts. Glucagon and insulin levels in the venous outflow from the graft and in the systemic circulation were determined during the first 60 minutes after transplantation. Two of the dogs were subjected to L-arginine stimulation 5 days after transplantation and the glucagon levels in the venous outflow from th
Suspensions of dispersed islet cells were prepared by shaking collagenaseisolated pancreatic islets of ob ob-mice in Ca2+-free buffer. The dispersed cells exhibited a glucose uptake with stereospecificity for the d isomer and concentrated Rb+ about 30-fold from a medium containing 70 μm RbCl. These results compare well with previous observations on unbroken islets and indicate that the dispersion
Insulin release and the content of cAMP were studied in microdissected pancreatic islets of non inbred ob/ob (obese) mice. In the absence of 3 isobutyl 1 methylxanthine, a phosphodiesterase inhibitor, 20 mM glucose had no effect on cAMP save a very small initial rise detectable by a freeze stop perifusion technique only. However, combined with this methylxanthine, 20 mM glucose produced significan
The effects of an electron dense thiol reagent, Hg phenylazo ferritin, on β cell ultrastructure was studied in cell suspensions from mouse pancreatic islets. This type of thiol reagent induced severe damage in the β cells. The ferritin particles were traced with the electron microscope and found to be bound to the plasma membrane as well as to various cytoplasmic structures. The β cell cytotoxic a
Mouse pancreatic islets microdissected free from, or surrounded by, exocrine cells were used to study the effects of secretin and choleeystokinm-pancreozymin on insulin release. Both secretin and cholecystokinin-pancreozymin potentiated glucose-stimulated insulin release regardless of whether exocrine cells were present. The results fail to support the idea that the presence of the exocrine parenc
Pancreatic islets, isolated from the pancreas of obese-hyperglycemic mice, were used to prepare free islet cells in suspension. Batches of 100 islets were disrupted by mechanical shaking for 10 sec in a Ca2+-free HEPES-buffered Krebs-Ring'er medium containing 1 mM EGTA. From 200-500 islets, about 2.2×106 cells could be obtained in suspension, corresponding to a yield of roughly 55% as calculated f
Body acceleration due to heartbeat-induced reaction forces can be measured as mobile phone accelerometer (m-ACC) signals. Our aim was to test the feasibility of using m-ACC to detect changes induced by stress by ultra-short heart rate variability (USV) indices (standard deviation of normal-to-normal interval-SDNN and root mean square of successive differences-RMSSD). Sixteen healthy volunteers wer
Mice, 7-8-mo old, of the C57BL/KsJ-db strain and homozygotic for the mutant gene db, exhibited marked hyperglycemia and moderately elevated serum insulin levels. Light and electron microscopy provided evidence of a slightly decreased proportion of β cells in the pancreatic islets, irregular islet architecture with intraislet ducts, and degenerative as well as hypertrophic changes in the individua
d-Glyceraldehyde stimulated the release of insulin from pancreatic islets of Umeå- ob ob-mice whether or not glucose was present in the medium. Like the action of glucose, that of d-glyceraldehyde was biphasic in time, exhibited a sigmoidal dose-response relationship, was potentiated by theophylline, arginine, iodoacetamide, or l-glyceraldehyde, and was inhibited by epinephrine, 2,4-dinitrophenol,
Pancreatic islets rich in β cells and microdissected from noninbred ob/ob mice contained about 15 pmol sialic acid per μg dry weight as determined by a microversion of the Warren thiobarbituric acid method. Removal of half of the sialic acid by treatment with Clostridium perfringens neuraminidase had no effect on the islet content of insulin, moderately inhibited glucose oxidation, and resulted in
Glucose (20 mM) released insulin from pancreatic islets of C57BL 6J- db2J db2J mice, the response being potentiated by 1 mM 3-isobutyl-1-methylxanthine. Islets of C57BL KsJ- db db mice failed to respond to glucose and released only little insulin when challenged with both glucose and methylxanthine. After incubation with 0 or 20 mM glucose alone the islet content of adenosine 3′:5′-cyclic monophos
Chloromercuribenzene-p-sulphonic acid (0.1 mM) or 5,5′-dithiobis-(2-nitrobenzoic acid) (1 mM) alone had no effect on cyclic AMP in microdissected pancreatic islets of non-inbred ob/ob mice. In the presence of 1 mM 3-isobutyl-1-methylxanthine, the mercurial increased and the disulphide decreased the cyclic AMP content. Both sulphydryl reagents stimulated insulin release whether 3-isobutyl-1-methylx
Treatment of pancreatic islets from ob/ob-mice with bacterial neuraminidase (0.8 to 40 mU/ml) resulted in a significant decrease of the sialic acid content and of the secretory response to glucose. The inhibitory effect on the glucose stimulated insulin release was reproduced with different batches of neuraminidase from Clostridium perfringens and Vibrio cholera. Treatment with neuraminidase affec
