Tri-modal In vivo Imaging of Pancreatic Islets Transplanted Subcutaneously in Mice.

PURPOSE: Transplantation of pancreatic islets (PIs) is a promising therapeutic approach for type 1 diabetes. The main obstacle for this strategy is that the outcome of islet engraftment depends on the engraftment site. It was our aim to develop a strategy for using non-invasive imaging techniques to assess the location and fate of transplanted PIs longitudinally in vivo. PROCEDURES: In order to overcome the limitations of individual imaging techniques and cross-validate findings by different modalities, we have combined fluorine magnetic resonance imaging (F-19 MRI), fluorescence imaging (FLI), and bioluminescent imaging (BLI) for studying subcutaneously transplanted PIs and beta cell-like cells (INS-1E cell line) in vivo. We optimized the transduction (using lentiviral vectors) and labeling procedures (using perfluoro crown ether nanoparticles with a fluorescence dye) for PIs and INS-1E cell imaging. RESULTS: The feasibility of using the proposed imaging methods for PI assessment was demonstrated both in vitro and in vivo. Our data suggested that F-19 MRI is suitable for high-resolution localization of transplanted cells and PIs; FLI is essential for confirmation of contrast localization by histology; and BLI is a reliable method to assess cell viability and survival after transplantation. No significant side effects on cell viability and function have been observed. CONCLUSIONS: The proposed tri-modal imaging platform is a valuable approach for the assessment of engrafted PIs in vivo. It is potentially suitable for comparing different transplantation sites and evaluating novel strategies for improving PI transplantation technique in the future.

Improved Labeling of Pancreatic Islets Using Cationic Magnetoliposomes.

Pancreatic islets (PIs) transplantation is an alternative approach for the treatment of severe forms of type 1 diabetes (T1D). To monitor the success of transplantation, it is desirable to follow the location of engrafted PIs non-invasively. In vivo magnetic resonance imaging (MRI) of transplanted PIs is a feasible cell tracking method; however, this requires labeling with a suitable contrast agent prior to transplantation. We have tested the feasibility of cationic magnetoliposomes (MLs), compared to commercial contrast agents (Endorem and Resovist), by labeling insulinoma cells and freshly isolated rat PIs. It was possible to incorporate Magnetic Ressonance (MR)-detectable amounts of MLs in a shorter time (4 h) when compared to Endorem and Resovist. MLs did not show negative effects on the PIs' viability and functional parameters in vitro. Labeled islets were transplanted in the renal sub-capsular region of healthy mice. Hypointense contrast in MR images due to the labeled PIs was detected in vivo upon transplantation, while MR detection of PIs labeled with Endorem and Resovist was only possible after the addition of transfection agents. These findings indicate that MLs are suitable to image PIs, without affecting their function, which is promising for future longitudinal pre-clinical and clinical studies involving the assessment of PI transplantation.

Comparison of different compressed sensing algorithms for low SNR 19 F MRI applications-Imaging of transplanted pancreatic islets and cells labeled with perfluorocarbons.

Transplantation of pancreatic islets is a possible treatment option for patients suffering from Type I diabetes. In vivo imaging of transplanted islets is important for assessment of the transplantation site and islet distribution. Thanks to its high specificity, the absence of intrinsic background signal in tissue and its potential for quantification, 19 F MRI is a promising technique for monitoring the fate of transplanted islets in vivo. In order to overcome the inherent low sensitivity of 19 F MRI, leading to long acquisition times with low signal-to-noise ratio (SNR), compressed sensing (CS) techniques are a valuable option. We have validated and compared different CS algorithms for acceleration of 19 F MRI acquisition in a low SNR regime using pancreatic islets labeled with perfluorocarbons both in vitro and in vivo. Using offline simulation on both in vitro and in vivo low SNR fully sampled 19 F MRI datasets of labeled islets, we have shown that CS is effective in reducing the image acquisition time by a factor of three to four without seriously affecting SNR, regardless of the particular algorithms used in this study, with the exception of CoSaMP. Using CS, signals can be detected that might have been missed by conventional 19 F MRI. Among different algorithms (SPARSEMRI, OMMP, IRWL1, Two-level and CoSAMP), the two-level l1 method has shown the best performance if computational time is taken into account. We have demonstrated in this study that different existing CS algorithms can be used effectively for low SNR 19 F MRI. An up to fourfold gain in SNR/scan time could be used either to reduce the scan time, which is beneficial for clinical and translational applications, or to increase the number of averages, to potentially detect otherwise undetected signal when compared with conventional 19 F MRI acquisitions. Potential applications in the field of cell therapy have been demonstrated.

In vivo and ex vivo 19-fluorine magnetic resonance imaging and spectroscopy of beta-cells and pancreatic islets using GLUT-2 specific contrast agents.

The assessment of the ß-cell mass in experimental models of diabetes and ultimately in patients is a hallmark to understand the relationship between reduced ß-cell mass/function and the onset of diabetes. It has been shown before that the GLUT-2 transporter is highly expressed in both ß-cells and hepatocytes and that D-mannoheptulose (DMH) has high uptake specificity for the GLUT-2 transporter. As 19-fluorine MRI has emerged as a new alternative method for MRI cell tracking because it provides potential non-invasive localization and quantification of labeled cells, the purpose of this project is to validate ß-cell and pancreatic islet imaging by using fluorinated, GLUT-2 targeting mannoheptulose derivatives (19 FMH) both in vivo and ex vivo. In this study, we confirmed that, similar to DMH, 19 FMHs inhibit insulin secretion and increase the blood glucose level in mice temporarily (approximately two hours). We were able to assess the distribution of 19 FMHs in vivo with a temporal resolution of about 20 minutes, which showed a quick removal of 19 FMH from the circulation (within two hours). Ex vivo MR spectroscopy confirmed a preferential uptake of 19 FMH in tissue with high expression of the GLUT-2 transporter, such as liver, endocrine pancreas and kidney. No indication of further metabolism was found. In summary, 19 FMHs are potentially suitable for visualizing and tracking of GLUT-2 expressed cells. However, current bottlenecks of this technique related to the quick clearance of the compound and relative low sensitivity of 19 F MRI need to be overcome. Copyright © 2016 John Wiley & Sons, Ltd.

Vitamin D supplementation ameliorates hypoinsulinemia and hyperglycemia in static magnetic field-exposed rat.

The purpose of this study is to evaluate the effects of vitamin D supplementation on glucose and lipid metabolism in static magnetic field (SMF)-exposed rats. Rats exposed to SMF (128 mT; 1 h/day) during 5 consecutive days showed an increase in plasma glucose level and a decrease in plasma insulin concentration. By contrast, the same treatment failed to alter body weight and plasmatic total cholesterol, high-density lipoprotein (HDL)-cholesterol, low-density lipoprotein (LDL)-cholesterol, and triglyceride levels. Interestingly, supplementation with vitamin D (1,600 IU/100 g, per os) corrected and restored glycemia and insulinemia in SMF-exposed rats. The same treatment had no effects on lipid metabolism.

Vitamin E prevents glucose metabolism alterations induced by static magnetic field in rats.

In the present study, we investigate the effects of a possible protective role of vitamin E (vit E) or selenium (Se) on glucose metabolism disruption induced by static magnetic field (SMF) in rats. Rats have been exposed to SMF (128 mT, 1 h/day during 5 days). Our results showed that SMF failed to alter body weight and relative liver weight. Our data demonstrated that exposure to SMF increased (+21 %) blood glucose level and caused a decrease (-15 %) in liver glycogen content. Moreover, the same treatment induced a reduction of pancreatic islet area. Interestingly, supplementation with vit E (DL α-tocopherol acetate, 150 mg/kg per os during 5 days) prevented alterations induced by SMF on glucose metabolism and liver glycogen content, whereas supplementation with Se (Na2SeO3, 0.20 mg/l, in drinking water for 4 weeks) restored only hepatic glycogen contents. By contrast, both vit E and Se failed to correct the area of pancreatic islets.

Insulinotropic action of Citrullus colocynthis seed extracts in rat pancreatic islets.

The present study aimed to investigate the direct in vitro effects of several distinct Citrullus colocynthis seed extracts on glucose-stimulated insulin release from pancreatic islets isolated from rats. Six extracts were tested, a crude aqueous, defatted aqueous, ethyl acetate, H2O-methanol and n-butanol extract and an extract containing a major component (fraction A) identified by gel chromatography in the ethyl acetate, n-butanol and H2O-methanol extracts. Under selected experimental conditions, the majority of extracts exhibited a positive insulinotropic action, at least when tested in the presence of 8.3 mM D-glucose. The concentration-response correlation observed with distinct extracts revealed the participation of distinct chemical compounds, including compounds with an inhibitory insulinotropic potential, in the modulation of the insulin secretory response to D-glucose. The results of the present study are relevant for further investigations which aim to identify compounds exhibiting positive insulinotropic actions. These agents may be suitable for the treatment of human diabetic subjects.

Protective action of Citrullus colocynthis seed extracts against the deleterious effect of streptozotocin on both in vitro glucose-stimulated insulin release from rat pancreatic islets and in vivo glucose homeostasis.

Citrullus colocynthis extracts improve glucose homeostasis in alloxan- or streptozotocin (STZ)-induced diabetic rats. Little is known, however, regarding the protective effect of these extracts against the ß-cytotoxic action of STZ. In the present study, an H2O-methanol extract was found to suppress the inhibition of glucose-stimulated insulin secretion by STZ in rat-isolated pancreatic islets. Similarly, when an aqueous extract from Citrullus colocynthis seeds was injected daily for 21 days prior to STZ administration, the perturbation of glucose homeostasis otherwise generated by the ß-cytotoxic agent was minimized in rats.

Uptake and efflux of 3-O-methyl-D-glucose in rat parotid cells.

In the framework of recent investigations on the regulation of D-glucose production by salivary glands, the aim of the present study was to compare the uptake of 3-O-[14C]methyl-D-glucose by rat parotid cells over a 6-min incubation period at 37°C to its efflux from prelabelled parotid cells, also incubated for 6 min at 37°C. It was first assessed that the intracellular 3HOH water space, whether expressed in absolute terms or relative to the total 3HOH distribution space, is not significantly different between parotid cells obtained from either control rats or streptozotocin-induced diabetic rats. In the control rats, the uptake of 3-O-[14C]methyl-D-glucose corresponded, following correction for extracellular contamination, to a mean distribution space of 0.44±0.05 nl/103 cells, representing 29.8±3.4% of the intracellular water space. The efflux of 3-O-[14C]methyl-D-glucose from prelabelled parotid cells, expressed relative to their initial radioactive content, averaged 82.9±4.8 and 84.1±2.5% in control and diabetic rats, respectively. These findings suggest that the increased production of salivary D-glucose in diabetic subjects may be attributable to hyperglycemia, rather than to any major perturbation of the intrinsic processes involved, at least in parotid cells, in hexose handling.

The metabolic syndrome of fructose-fed rats: effects of long-chain polyunsaturated ω3 and ω6 fatty acids. V. Post-mortem findings.

The present study deals with the possible effects of dietary ω3 and ω6 fatty acids upon the metabolic syndrome found in rats exposed for 8 weeks to a diet containing 64% (w/w) D-fructose instead of starch. Fructose-fed rats were found to display a modest increase in plasma albumin and protein concentration and more pronounced increases in plasma urea, creatinine, phospholipids, triglycerides and cholesterol concentrations, glycated hemoglobin concentration and liver contents of cholesterol, triglycerides and phospholipids. The plasma concentrations of HDL-cholesterol, calcium and iron were decreased, however, in the fructose-fed rats. In general, the partial substitution of sunflower oil by either safflower oil or salmon oil opposed the metabolic perturbations otherwise associated with the fructose-induced metabolic syndrome in the fructose-fed rats, with salmon oil demonstrating particular efficacy. Consideration is given to the possible biological determinants of these perturbations and their attenuation in rats exposed to safflower or salmon oil.

Glucose transport by acinar cells in rat parotid glands.

BACKGROUND/AIMS: Salivary glucose is often considered as being from glandular origin. Little information is available, however, on the possible role of glucose transporters in the secretion of the hexose by salivary glands. The major aim of the present study was to investigate the expression and localization of several distinct glucose transporters in acinar cells of rat parotid glands. METHODS: Quantitative real-time PCR analysis, immunohistochemistry and western blotting techniques were used to assess the presence of SGLT1, GLUT1, GLUT2 and GLUT4 in acinar cells of rat parotid glands. RESULTS: Quantitative real-time PCR documented the expression of SGLT1 and GLUT1 in parotid tissues, with a much lower level of GLUT4 mRNA and no expression of GLUT2 mRNA. Western blot analysis revealed the presence of SGLT1, GLUT1 and GLUT4 proteins, but not GLUT2 proteins in the parotid extract. Immunohistochemistry confirmed these findings. SGLT1 was specifically located at the baso-lateral membrane, co-localizing with Na(+)/K(+) ATPase. GLUT1 was found both at the baso-lateral and apical level. GLUT4 appeared to be also located at the baso-lateral level. However, too little GLUT4 was present to allow co-localization labeling. CONCLUSION: Based on these findings, a model is proposed for the transport of glucose into the acinar cells and thereafter into the acinar lumen.

A new role for aquaporin 7 in insulin secretion.

UNLABELLED: BACGROUNS/AIMS: Several insulinotropic agents were recently reported to cause ß-cell swelling. The possible participation of AQP7 to water transport was investigated in AQP7(+/+) or AQP7(-/-) mice. METHODS: Aquaporin expression, insulin secretion, cell swelling and electrical activity were investigated in pancreatic islets. RESULTS: RT-PCR revealed the expression of AQP5 and AQP8 mRNA. Double immunofluorescent labeling indicated their presence in ß-cells. Whilst basal insulin release from isolated pancreatic islets incubated at 2.8 mM D-glucose did not differ between AQP7(+/+) or AQP7(-/-) mice, the secretion of insulin evoked by the omission of 50 mM NaCl, the substitution of 50 mM NaCl by 100 mM glycerol or a rise in D-glucose concentration to 8.3 mM and 16.7 mM was severely impaired in the islets from AQP7(-/-) mice. Yet, exposure of ß-cells to either the hypotonic medium or a rise in D-glucose concentration caused a similar degree of swelling and comparable pattern of electrical activity in cells from AQP7(+/+) and AQP7(-/-) mice. Both the cell swelling and change in membrane potential were only impaired in AQP7(-/-) cells when exposed to 50 mM glycerol. CONCLUSION: It is proposed, therefore, that AQP7 may, directly or indirectly, play a role at a distal site in the exocytotic pathway.

Perturbation of glycerol metabolism in hepatocytes from n3-PUFA-depleted rats.

Second generation n3-PUFA-depleted rats represent a good animal model of metabolic syndrome as they display several features of the disease such as liver steatosis, visceral obesity and insulin resistance. The goal of our study was to investigate the influence of n3-PUFA deficiency on hepatic glycerol metabolism. Aquaglyceroporin 9 (AQP9) allows hepatic glycerol transport and consequently contributes to neoglucogenesis. AQP9 knockout mice display hypertriacyl-glycerolemia, one of the hallmarks of the metabolic syndrome. Our data show reduced AQP9 expression at the protein level in n3-PUFA-depleted rats, without any changes at the mRNA levels. [U-¹4C]glycerol uptake was increased in hepatocytes from n3-PUFA-depleted animal cells. The apparent discrepancy between decreased AQP9 protein expression, and increased [U-¹4C]glycerol uptake could be explained by an observed increase in glycerol kinase activity.

Intermittent fasting modulation of the diabetic syndrome in streptozotocin-injected rats.

This study investigates the effects of intermittent overnight fasting in streptozotocin-induced diabetic rats (STZ rats). Over 30 days, groups of 5-6 control or STZ rats were allowed free food access, starved overnight, or exposed to a restricted food supply comparable to that ingested by the intermittently fasting animals. Intermittent fasting improved glucose tolerance, increased plasma insulin, and lowered Homeostatis Model Assessment index. Caloric restriction failed to cause such beneficial effects. The ß-cell mass, as well as individual ß-cell and islet area, was higher in intermittently fasting than in nonfasting STZ rats, whilst the percentage of apoptotic ß-cells appeared lower in the former than latter STZ rats. In the calorie-restricted STZ rats, comparable findings were restricted to individual islet area and percentage of apoptotic cells. Hence, it is proposed that intermittent fasting could represent a possible approach to prevent or minimize disturbances of glucose homeostasis in human subjects.

The metabolic syndrome of fructose-fed rats: effects of long-chain polyunsaturated ω3 and ω6 fatty acids. III. Secretory behaviour of isolated pancreatic islets.

In the present study, rats were exposed from the 8th week after birth and for the ensuing 8 weeks to diets containing either starch or fructose (64% w/w) and sunflower oil (5%). Two further groups of rats were exposed to the fructose-containing diet with substitution of part (1.6%) of the sunflower diet by an equal amount of either salmon oil rich in long-chain polyunsaturated ω3 fatty acids or safflower oil reach in long-chain polyunsaturated ω6 fatty acids. The insulin content of the islets and their secretory response to D-glucose (5.6, 8.3 and 16.7 mM), to the combination of D-glucose (5.6 mM) and D-fructose (10.0 mM) and to 2-ketoisocaproate (10.0 mM) were then measured. In the sunflower oil-fed rats, the dietary substitution of starch by fructose decreased basal insulin output, lowered the apparent Km for the insulinotropic action of D-glucose and altered the insulinotropic efficiency of the latter hexose relative to that of other nutrients. Some of these secretory perturbations were opposed by the enrichment of the diet in long-chain polyunsaturated fatty acids, especially ω3 fatty acids. It is proposed that these changes in B-cell secretory behaviour may be linked, in part at least, to both the apparent caloric efficiency of each diet, and hence to the regulation of the islet content in endogenous nutrients, and to alteration of insulin sensitivity considered as a major feature of the present animal model of metabolic syndrome.

Dietary sardine protein lowers insulin resistance, leptin and TNF-α and beneficially affects adipose tissue oxidative stress in rats with fructose-induced metabolic syndrome.

The present study aims at exploring the effects of sardine protein on insulin resistance, plasma lipid profile, as well as oxidative and inflammatory status in rats with fructose-induced metabolic syndrome. Rats were fed sardine protein (S) or casein (C) diets supplemented or not with high-fructose (HF) for 2 months. Rats fed the HF diets had greater body weight and adiposity and lower food intake as compared to control rats. Increased plasma glucose, insulin, HbA1C, triacylglycerols, free fatty acids and impaired glucose tolerance and insulin resistance was observed in HF-fed rats. Moreover, a decline in adipose tissues antioxidant status and a rise in lipid peroxidation and plasma TNF-α and fibrinogen were noted. Rats fed sardine protein diets exhibited lower food intake and fat mass than those fed casein diets. Sardine protein diets diminished plasma insulin and insulin resistance. Plasma triacylglycerol and free fatty acids were also lower, while those of α-tocopherol, taurine and calcium were enhanced as compared to casein diets. Moreover, S-HF diet significantly decreased plasma glucose and HbA1C. Sardine protein consumption lowered hydroperoxide levels in perirenal and brown adipose tissues. The S-HF diet, as compared to C-HF diet decreased epididymal hydroperoxides. Feeding sardine protein diets decreased brown adipose tissue carbonyls and increased glutathione peroxidase activity. Perirenal and epididymal superoxide dismutase and catalase activities and brown catalase activity were significantly greater in S-HF group than in C-HF group. Sardine protein diets also prevented hyperleptinemia and reduced inflammatory status in comparison with rats fed casein diets. Taken together, these results support the beneficial effect of sardine protein in fructose-induced metabolic syndrome on such variables as hyperglycemia, insulin resistance, hyperlipidemia and oxidative and inflammatory status, suggesting the possible use of sardine protein as a protective strategy against insulin resistance and related situations.

The metabolic syndrome of fructose-fed rats: effects of long-chain polyunsaturated ω3 and ω6 fatty acids. II. Time course of changes in food intake, body weight, plasma glucose and insulin concentrations and insulin resistance.

The time course for changes in food intake, body weight, plasma glucose and insulin concentrations and HOMA index was monitored over a period of 8 weeks in rats exposed from the 8th week after birth to diets containing either starch or fructose and sunflower oil. In two further groups of rats exposed to the fructose-rich diet part of the sunflower oil was substituted by either salmon oil rich in long-chain polyunsaturated ω3 fatty acids or safflower oil rich in long-chain polyunsaturated ω6 fatty acids. Despite lower food intake, the gain in body weight was higher in fructose-fed rats than in starch-fed rats. The supplementation of the fructose-rich diet by either ω3 or ω6 fatty acids lowered both food intake and body weight gain. The measurements of plasma glucose and insulin concentrations, HOMA index and insulinogenic index performed after overnight starvation were in fair agreement with those recorded at the occasion of an intraperitoneal glucose tolerance test, with higher values for plasma glucose concentration and HOMA index in the fructose-fed rats exposed to the sunflower oil (with or without enrichment with ω6 fatty acids) than in the starch-fed rats exposed to the sunflower oil or fructose-fed rats exposed to a diet enriched with ω3 fatty acids. Such was also the case for the measurements of glycated albumin at sacrifice. Moreover, the insulinogenic index was lower in the fructose-fed rats with or without dietary enrichment in ω6 fatty acids than in the fructose-fed rats with dietary enrichment in ω3 fatty acids. The elucidation of the biochemical determinants of the later difference requires further investigations in isolated pancreatic islets.

The metabolic syndrome of fructose-fed rats: effects of long-chain polyunsaturated ω3 and ω6 fatty acids. IV. D-glucose metabolism by isolated pancreatic islets.

The major aim of the present study was to search for changes of D-glucose metabolism in isolated pancreatic islets possibly involved in the alteration of their secretory response to the hexose, as observed when comparing rats exposed for 8 weeks to diets containing either starch and sunflower oil or fructose and sunflower oil, as well as rats exposed to diets containing fructose, sunflower oil and either salmon oil or safflower oil. The substitution of starch by fructose in the diet affected unfavourably D-glucose phosphorylation by the isolated islets. In the fructose-fed rats, there was a close parallelism between D-[5-³H]glucose utilization and the dietary ω3/ω6 fatty acid ratio. There was little to distinguish, however, between the four groups of rats in terms of D-[U-¹4C]glucose oxidation. The paired ratio between D-[U-¹4C]glucose oxidation and D-[5-³H]glucose utilization, which always increased as the concentration of the hexose was raised from 2.8 to 8.3 and 16.7 mM, was tightly related, in the fructose-fed rats, to the HOMA index for insulin resistance.

(19)F-heptuloses as tools for the non-invasive imaging of GLUT2-expressing cells.

Suitable analogs of d-mannoheptulose are currently considered as possible tools for the non-invasive imaging of pancreatic islet insulin-producing cells. Here, we examined whether (19)F-heptuloses could be used for non-invasive imaging of GLUT2-expressing cells. After 20 min incubation, the uptake of (19)F-heptuloses (25 mM) by rat hepatocytes, as assessed by (19)F NMR spectroscopy, ranged from 0.50 (1-deoxy-1-fluoro-d-mannoheptulose and 3-deoxy-3-fluoro-d-mannoheptulose) to 0.25 (1,3-dideoxy-1,3-difluoro-d-mannoheptulose) and 0.13 (1-deoxy-1-fluoro-d-glucoheptulose, 3-deoxy-3-fluoro-d-glucoheptulose and 1,3-dideoxy-1,3-difluoro-d-glucoheptulose) µmol per 3×10(6)cells. (19)F MRI experiments also allowed the detection of 1-deoxy-1-fluoro-d-mannoheptulose in rat hepatocytes. All three (19)F-mannoheptuloses cited above, as well as 7-deoxy-7-fluoro-d-mannoheptulose and 1-deoxy-1-fluoro-d-glucoheptulose inhibited insulin release evoked in rat isolated pancreatic islets by 10mM d-glucose to the same extent as that observed with an equivalent concentration (10mM) of d-mannoheptulose, while 3-deoxy-3-fluoro-d-glucoheptulose and 1,3-dideoxy-1,3-difluoro-d-glucoheptulose (also 10mM) were less potent than d-mannoheptulose in inhibiting insulin release. The 1-deoxy-1-fluoro-d-mannoheptulose and 3-deoxy-3-fluoro-d-mannoheptulose only marginally affected INS-1 cell viability. These findings are compatible with the view that selected (19)F-heptuloses may represent suitable tools for the non-invasive imaging of hepatocytes and insulin-producing cells by (19)F MRI.

The metabolic syndrome of fructose-fed rats: effects of long-chain polyunsaturated ω3 and ω6 fatty acids. I. Intraperitoneal glucose tolerance test.

The present series of experiments aim mainly at investigating the possible influence of changes in the com-position of dietary lipids (sunflower oil, salmon oil, safflower oil) upon the metabolic syndrome found in rats exposed to a fructose-rich diet. For purpose of comparison, a control group of rats received the sunflower oil diet with substitution of fructose by starch. An intraperitoneal glucose tolerance test, performed after overnight starvation fifty days after the start of the experiments at the 6th week after birth, indicated, as expected, impaired tolerance to glucose and deterioration of insulin sensitivity (HOMA index), without changes in the insulinogenic index, when comparing the fructose-fed rats to the starch-fed rats both exposed to the sunflower oil diet. In the fructose-fed rats, enrichment of the diet by long-chain polyunsaturated ω3 fatty acids supplied by salmon oil, a modest improvement of insulin sensitivity was opposed, in term of glucose homeostasis, by a decreased secretory response to glucose of insulin-producing cells. Last, in the fructose-fed rats, the partial substitution of sunflower oil by safflower oil rich in long-chain polyunsaturated ω6 fatty acids further deteriorated glucose homeostasis, with a higher mean HOMA index and a severe decrease of the insulinogenic index. These findings justify further investigations on such items as the time course for changes in metabolic and hormonal variables and both the metabolic and secretory responses of isolated pancreatic islets to selected nutrient secretagogues.