Expression of glucose transporter 4 in the human pancreatic islet of Langerhans

Glucose transporter 4 (GLUT4) is the main insulin-responsive glucose transporter in skeletal muscle and adipose tissue of human and rodent, and is translocated to the plasma membrane in response to insulin. GLUT2 is well known as the main glucose transporter in pancreatic islets and could highly regulate glucose-stimulated insulin secretion by B-cells as a glucose sensor. We confirmed the presence of GLUT4 mRNA and GLUT4 protein in pancreas in the human. Indirect immunohistochemistry showed that the pancreatic islets of human and rat were conspicuously labeled by anti-GLUT4 antibody. The presence of placental leucine aminopeptidase (P-LAP), a homologue of insulin-regulated aminopeptidase (IRAP), was also shown in the human pancreatic islet. IRAP/P-LAP is thought to be involved in glucose metabolism. This study provides the first evidence that GLUT4 is present in human and rat pancreatic islets and may suggest its specific role in glucose homeostasis in conjunction with IRAP/P-LAP.     

Inhibition by Celiptium of the fetal thymidine kinase synthesis induced by estrogens in the rat uterus

Celiptium (Ce) is an antitumor drug used in the therapy of breast carcinomas, which are to a large extent dependent on estrogens. We have studied the effect of Ce on some proteins induced by estradiol (E2) in the rat uterus. It was observed that Ce administered at the same time or before E2, was able to inhibit the induction by E2 of fetal thymidine kinase (TK-F), of creatine kinase of brain-type (CK-BB) and of progesterone receptor (PR). When Ce was given after E2, its inhibitory effects were less evident. Results seemed to indicate that Ce could bind the acceptor sites for E2 receptor and thus inhibit the activity of E2-regulated genes. This assumption was corroborated by the fact that Ce did not modify the activity of enzymes not submitted to E2 regulation.     

Different metabolic responses in alpha-, beta-, and delta-cells of the islet of Langerhans monitored by redox confocal microscopy

Blood glucose homeostasis is mainly achieved by the coordinated function of pancreatic alpha-, beta-, and delta-cells, which secrete glucagon, insulin, and somatostatin, respectively. Each cell type responds to glucose changes with different secretion patterns. Currently, considerable information can be found about the signal transduction mechanisms that lead to glucose-mediated insulin release in the pancreatic beta-cell, mitochondrial activation being an essential step. Increases in glucose stimulate the mitochondrial metabolism, activating the tricarboxylic acid cycle and raising the source of redox electron carrier molecules needed for respiratory ATP synthesis. However, little is known about the glucose-induced mitochondrial response of non-beta-cells and its role in the stimulus-secretion coupling process. This limited information is probably a result of the scarcity of these cells in the islet, the lack of identification patterns, and the technical limitations of conventional methods. In this study, we used flavin adenine dinucleotide redox confocal microscopy as a noninvasive technique to specifically monitor mitochondrial redox responses in immunoidentified alpha-, beta-, and delta-cells in freshly isolated intact islets and in dispersed cultured cells. We have shown that glucose provokes metabolic changes in beta- and delta-cell populations in a dose-dependent manner. Conversely, no significant responses were observed in alpha-cells, despite the sensitivity of their metabolism to drugs acting on the mitochondrial function, and their intact ability to develop Ca2+ signals. Identical results were obtained in islets and in cultures of dispersed cells. Our findings indicate metabolic differences in glucose utilization among the alpha-, beta-, and delta-cell populations, which might be important in the signal transduction events that lead to hormone release.     

Effect of estrogens on dmba induced breast tumors

Evidence is presented for the anti-carcinogenic activity of estriol when this hormone is administered to 50–55 day old SD rats prior to treatment with DMBA. In control and estradiol treated animals, 85–100% of animals developed breast tumors regardless of dose of steroid. Low doses of estriol (1 μg/day) offered some protection against tumorigenesis but at levels exceeding 10μg/day only 20–35% of animals developed tumors even when observed for 180 days following DMBA ingestion. Furthermore, the mean time for tumor production is greatly increased in the presence of E₃. Short term (2 weeks) administration offered no protection against tumor formation, suggesting that the effect of estriol probably relates to the promotion stage of tumorigenesis.     

Inflammatory mediators expressed in human islets of Langerhans: implications for islet transplantation

Expression of immune modulating mediators in human Islets of Langerhans could have important implications for development of autoimmunity in type 1 diabetes and influence the outcome of clinical islet transplantation. Islets obtained from five donors were analyzed at various times after isolation using cDNA array technology. The Atlas Human Cytokine/Receptor and Hematology/Immunology nylon membranes representing 268 genes and 406, respectively, were used and the relative expression of each gene analyzed. Of the 51 gene products identified, high mRNA expression of MCP-1, MIF, VEGF, and thymosin beta-10 was detected in all islet samples. IL-8, IL-1-beta, IL-5R, and INF-gamma antagonist were expressed in islets cultured for 2 days. IL-2R was expressed in islets cultured for more than 6 days. In conclusion, several inflammatory mediators were expressed in isolated islets, particularly at an early stage after isolation, indicating that a few days of culture could be beneficial for the outcome of islet transplantation.     

Signaling by estrogens

By regulating activities and expression levels of key signaling molecules, estrogens control mechanisms that are responsible for crucial cellular functions. Ligand binding to estrogen receptor (ER) leads to conformational changes that regulate the receptor activity, its interaction with other proteins and DNA. In the cytoplasm, receptor interactions with kinases and scaffolding molecules regulate cell signaling cascades (extranuclear/nongenomic action). In the nucleus, estrogens control a repertoire of coregulators and other auxiliary proteins that are associated with ER, which in turn determines the nature of regulated genes and level of their expression (genomic action). The combination of genomic and nongenomic actions of estrogens ultimately confers the cell-type and tissue-type selectivity. Recent studies have revealed some important new insights into the molecular mechanisms underlying ER action, which may help to explain the functional basis of existing selective ER modulators (SERMs) and provide evidence into how ER might be selectively targeted to achieve specific therapeutic goals. In this review, we will summarize some new molecular details that relate to estrogen signaling. We will also discuss some new strategies that may potentially lead to the development of functionally selective ER modulators that can separate between the beneficial, prodifferentiative effects in bone, the cardiovascular system and the CNS as well as the "detrimental," proliferative effects in reproductive tissues and organs.     

Correlation of islet size and biochemical parameters of isolated islets of Langerhans of rats.

[3H]-Leucine incorporation, insulin secretion, insulin content and DNA content of isolated islets of Langerhans of rats were determined. All these parameters are linearly correlated with the cross-sectional area of the isolated islets measured by means of an ocular micrometer.     

Hyperplasia of myoepithelial cells expressing calponin during atrophy of the rat parotid gland induced by duct ligation

The number and location of myoepithelial cells in the rat parotid gland submitted to surgical ligation of its main excretory duct were studied through immunohistochemical labelling for calponin. These cells were labelled by the streptavidin-biotin method using anti-calponin primary antibody, and their number was determined during each step of glandular atrophy, i.e., at time zero (control) and 1, 7, 15, 21, 30 and 60 days after ligation. Morphological analysis showed a gradual decrease and fibrosis of the glandular lobules accompanied by disappearance of the acini and the occurrence of duct-like structures. The expression of calponin was observed in all specimens analysed, being restricted to myoepithelial cells. Labelling revealed the distribution of myoepithelial cells around the acini and intercalated ducts in the control group and around duct-like structures later during the course of atrophy. Quantitative analysis demonstrated significant increase in the number of myoepithelial cells up to day 7 post-ligation, followed by gradual increases which, however, were not statistically significant. These results suggest that myoepithelial cells proliferate intensely up to day 7 post-ligation, an event that coincides with a higher rate of disappearance of acinar cells. After this period, the elevated number of cells observed at the end of the previous period is maintained.