[Growth and regeneration potential abnormalities of pancreatic beta-cells in GK rats, NIDDM mellitus model].
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Age Factors↗
Biomedical subjects
B Portha
Publications and source records attributed to B Portha.
Effect of gliclazide treatment on insulin secretion and beta-cell mass in non-insulin dependent diabetic Goto-Kakisaki rats.
The Goto-Kakisaki rat is a genetic non-overweight model of non-insulin-dependent diabetes mellitus. Adult Goto-Kakisaki rats exhibit a mild basal hyperglycaemia (11 mmol/l) with impaired glucose tolerance, elevated basal plasma insulin level, a failure of insulin release in response to glucose together with a 50% depletion of the total pancreatic beta-cell mass and insulin stores. We have examined the effects of long-term (4 weeks) gliclazide treatment on the severity of diabetes in adult male Goto-Kakisaki rats (10-12 weeks of age). Gliclazide was administered orally (10 mg/kg per day). Gliclazide-treated Goto-Kakisaki rats were evaluated against Wistar and untreated Goto-Kakisaki rats. In the gliclazide-treated Goto-Kakisaki rats, basal plasma glucose levels declined progressively reaching 8 mmol/l as a mean at the end of treatment, and their basal insulin levels decreased to values similar to those in non-diabetic Wistar rats. Despite their total pancreatic beta-cell remaining unaffected, their pancreatic insulin stores were twice increased, with a similar improvement of the insulin content per individual beta-cell. Furthermore, the glucose-stimulated insulin release as evaluated in vivo during an intravenous glucose tolerance-test was significantly improved (twice increased) in the gliclazide-treated Goto-Kakisaki rats. This was correlated with a modest but significant enhancement of the early phase of insulin release in vitro (isolated perfused pancreas), in response to glucose. However, the overall insulin response in vitro remained clearly defective with no reappearance of the late phase of insulin release. The in vitro response to arginine (which was basically amplified in the Goto-Kakisaki model) or to gliclazide were kept unchanged after the gliclazide treatment. In conclusion, chronic gliclazide does not exert any beta-cytotrophic effect, but improves beta-cell function in the adult Goto-Kakisaki rat as far as it lowers basal insulin release, increases beta-cell insulin stores, and increases the glucose-induced insulin release.
Administration, Oral↗
Impaired pancreatic beta cell function in the fetal GK rat. Impact of diabetic inheritance.
The Goto-Kakisaki (GK) rat is a genetic model of non-insulin-dependent diabetes. At 21.5 d of age we found that GK fetuses had an increased plasma glucose concentration, a decreased plasma insulin level, and a reduced pancreatic beta cell mass. To investigate the beta cell function during fetal life we used a hyperglycemic clamp protocol applied to the mothers, which allowed us to obtain a steady-state hyperglycemia in the corresponding fetuses. At variance, with Wistar (W) fetuses, plasma insulin concentration in GK fetuses did not rise in response to hyperglycemia. In contrast, GK fetal pancreas released insulin in response to glucose in vitro to the same extent as W fetal pancreas. Such a discrepancy between the in vivo and in vitro results suggests that the lack of pancreatic reactivity to glucose as seen in vivo is extrinsic to the fetal GK beta cell. Finally, the importance of gestational hyperglycemia was investigated by performing crosses between GK and W rats. Fetuses issued from crosses between W mother and GK father or GK mother and W father had a beta cell mass close to normal values and were still able to increase their plasma insulin levels in response to hyperglycemia in vivo. Our data suggest that hyperglycemia in utero does not influence the severity of the decrease of the beta cell mass or the lack of the insulin secretory response to glucose in the fetal GK rat. Moreover they indicate that conjunction of GK genes originating from both parents is necessary in order for these defects to be fully expressed.
Animals↗
Evaluation of dual-energy X-Ray absorptiometry for body-composition assessment in rats.
Recent developments in dual-energy X-ray absorptiometry (DXA) have rendered feasible the determination of whole-body composition in small laboratory animals by directly measuring fat, fat-free and mineral bone masses. Our aim was to evaluate this technique by cross-calibrating the DXA method with the carcass chemical analysis in a heterogeneous population of nondiabetic Wistar and diabetic GK rats (21 animals were used for precision error and reproducibility determinations and 26 were used for accuracy studies). We report that this technique is optimized for weights >200 g. The respective CV for lean mass, fat mass and percentage of fat mass determined in short-term or transversal studies was 1.1 +/- 0.1, 3.0 +/- 1.3 and 3. 1 +/- 0.4% (mean +/- SD) respectively. Further, this technique is valid for rats weighing from 130 to 200 g by using three successive scans. In longitudinal studies, daily calibrations significantly increased the percentage of fat mass CV to 6.6 +/- 3.3%, but it was significantly decreased to 3.0 +/- 2.7% by the use of triplicate scans. The accuracy for DXA was excellent in reference to the chemical extraction technique (r2 = 0.95 for percentage of fat mass, P < 0.0001), using an adjustment factor of 0.75 (limits of agreement between the two methods for percentage of fat mass = -1.7-2.3%). Mimicry of longitudinal changes in body composition with intraperitoneal injections of saline solution demonstrated a satisfactory detection of body component changes (</=2% of error for each final component analyzed, when increasing total lean mass by 11. 8%). We conclude that DXA is appropriate for rat whole-body composition determination, allowing reliable long-term follow-up of individual animals for the first time.
Absorptiometry, Photon↗
Altered activity of the autonomous nervous system as a determinant of the impaired beta-cell secretory response after protein-energy restriction in the rat.
Glucose-induced insulin secretion in vivo is known to be severely blunted in the rat as a consequence of protein-energy restriction starting early in life. We have recently reported in such malnourished rats (M rats) that the release of the counterregulatory hormones that defend against hypoglycemia was severely disturbed, and their plasma levels of epinephrine and norepinephrine were prominently increased. Knowing that the autonomic nervous system has the potential to play a major role in the control of insulin secretion in response to glucose in vivo, we therefore determined whether protein-energy restriction starting after weaning could alter sympathetic and/or parasympathetic nerve activities, and whether these changes could be responsible for the lack of response to glucose of their beta-cells in vivo. When tested in the basal postabsorptive state, the malnourished rats exhibited profound alterations of both parasympathetic and sympathetic nerve activities; the firing rates of the vagus nerve and the superior cervical ganglion were dramatically decreased and increased, respectively. Under the same conditions, insulin secretion in vivo in response to a glucose load (deltaI/deltaG) was severely decreased in M rats compared with that in control (C) rats. When evaluated after administration of acetylcholine, deltaI was amplified to the same extent in M rats as in C rats. After administration of the alpha2A-adrenergic agonist oxymetazoline, glucose-induced insulin release in M rats was not significantly affected, whereas it was sharply decreased in C rats. Finally, administration of yohimbine, an alpha2-adrenergic antagonist, partially restored the lack of reactivity of the beta-cells to glucose in the M rats, as deltaI/deltaG was amplified by 6-fold in the M group and by 3.3-fold in the C group. We conclude that protein-energy restriction starting early in life in rats brings about changes in the overall activity of the autonomic nervous system that, in turn, are responsible at least in part for the acquisition/maintenance of decreased beta-cell reactivity to glucose in vivo.
Acetylcholine↗
Pancreatic beta-cell regeneration after 48-h glucose infusion in mildly diabetic rats is not correlated with functional improvement.
We investigated the effect of glucose infusion on beta-cell regeneration in rats made mildly diabetic by a single injection of low dosage (35 mg/kg) streptozotocin (STZ). Nondiabetic (ND) and STZ rats were submitted to a 48-h glucose infusion (hyperglycemia approximately 22 mmol/l in both groups: ND and STZ hyperglycemic-hyperinsulinemic [ND HG-HI and STZ HG-HI rats]). Before infusion, beta-cell mass was 65% lower in STZ rats than in ND rats (2.0 +/- 0.02 vs. 5.5 +/- 0.6 mg), 1.6-fold increased in ND HG-HI rats (8.7 +/- 1.7 mg), and 2.7-fold increased in STZ HG-HI rats (5.4 +/- 0.9 mg). In ND HG-HI rats, beta-cell enlargement was related to an increase in beta-cell responsiveness to nutrient secretagogues both in vivo and in vitro, whereas in STZ HG-HI rats, no significant improvement in insulin secretion could be noticed. To determine the respective role of hyperglycemia and hyperinsulinemia on beta-cell area changes, ND and STZ rats were submitted to a 48-h hyperinsulinemic-euglycemic clamp. No modification of beta-cell mass was detected in either group. In conclusion, 48-h superimposed hyperglycemia was enough to restore beta-cell mass previously reduced by STZ injection. This effect seemed to be due to hyperglycemia rather than hyperinsulinemia alone. The data stress the dissociation between beta-cell regeneration and improvement in islet function in diabetic rats. Our model seems suitable for studying factors that can improve the plasticity and function of the pancreas in NIDDM.
Animals↗
Effects of dehydroepiandrosterone in rats injected with streptozotocin during the neonatal period.
Control rats and diabetic animals injected with streptozotocin during the neonatal period were either maintained on a standard diet or given access to food supplemented with dehydroepiandrosterone (DHEA, 0.2%) for 11 days before sacrifice. In both control and diabetic rats, DHEA feeding augmented the activity of the mitochondrial FAD-linked glycerophosphate dehydrogenase and cytosolic NADP-linked malate dehydrogenase in liver, but not so in either the parotid gland or pancreatic islets. DHEA lowered, in both control and diabetic rats, the ratio between D-glucose oxidation and utilization and the rate of insulin release in pancreatic islets exposed to a high concentration of D-glucose, as well as the insulin concentration and insulin/glucose ratio in plasma. These findings support the view that, in diabetes, DHEA, by increasing sensitivity to insulin, may allow islet B-cells to avoid the otherwise unfavorable consequences of chronic hyperactivity.
Administration, Oral↗
Transplantation of syngenic pancreatic islets into rats with streptozotocin induced non insulin dependent diabetes mellitus.
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Animals↗
Impaired development of pancreatic beta-cell mass is a primary event during the progression to diabetes in the GK rat.
In the endocrine pancreas of the GK rat, a genetic model of non-insulin-dependent diabetes mellitus (NIDDM), it is not clear whether the histopathological changes reported up to now are related to the pathogenesis of hyperglycaemia or whether they occur secondarily to metabolic alterations. Using GK rats from the Paris colony, our study chronicles for the first time the pathophysiologic changes that occur in the GK pancreas from the late fetal period (day 21.5) until adult age (18 weeks). As compared to Wistar controls, GK fetuses exhibited higher plasma glucose level, lower plasma insulin level and normal plasma glucagon level. Their pancreatic insulin content and the relative volume and the total mass of their beta cells were sharply decreased, representing only 23, 38 and 23% of control values, respectively. During the period from 4 days to 14 days after birth, GK neonates exhibited normal basal plasma glucose and glucagon levels despite decreased plasma insulin level. Their pancreatic insulin content represented only 31-40% of values found in the age-related control pancreases and their total beta-cell mass was only 35% on day 4, 30% on day 7 and 37% on day 14. The adult diabetic GK rats exhibited higher basal plasma glucose and insulin levels while their basal plasma glucagon level remained normal. Their pancreatic insulin content and the total beta-cell mass remained decreased, representing only 32% and 47% of control values, respectively. Moreover, the adult GK pancreases exhibited noticeable alteration in the architecture of the large islet subpopulation which displayed considerable fibrosis with clusters of beta cells widely separated from each other by strands of connective tissue. Concerning the development of alpha cells in the GK rats, their relative volume was found to be normal during fetal and early neonatal periods. It was found to be moderately decreased (representing 64-67% of corresponding control values) in 14-day-old neonates and adult GK rats. Our findings demonstrate that in the GK rat, the deficit of total beta-cell mass as observed in the adult animal is related to impaired beta-cell development. The restriction of the beta-cell mass must be considered as a primary and crucial event in the sequence leading to overt diabetes in this NIDDM model.
Age Factors↗
Alteration of the counterregulatory hormones in the conscious rat after protein-energy restriction.
We have recently reported that in rats submitted to protein-energy restriction early in life, an increased insulin efficiency upon the whole-body glucose utilization rate may be one reason for their chronic mild basal hypoglycaemia. However, the basis for their low plasma glucose level may also lie in the impaired activation of one or several of the counterregulatory hormones that prevent or correct hypoglycaemia. Our study was therefore designed to compare glucose counterregulatory mechanisms in restricted and control rats, both in the basal postabsorptive state and at controlled high plasma insulin level and standardized low glycaemic level (hypoglycaemic-hyperinsulinaemic glucose clamps performed in conscious rats). When tested in the basal postabsorptive state, the restricted rats exhibited prominent increases in the plasma levels of epinephrine (4.5 fold), norepinephrine (3.4 fold) and glucagon (1.7 fold). This was in the presence of significant decreases of plasma growth hormone and corticosterone levels (by 59 and 32%, respectively). With respect to the responses to acute severe hypoglycaemia (2.5 mmol/l), the glucagon, epinephrine and norepinephrine plasma levels in the restricted rats increased to values similar to those in controls. Also, the corticosterone level increased but remained significantly lower (p < 0.001) compared to the control response. The plasma growth hormone level was not significantly affected by acute hypoglycaemia in the restricted or in the control groups. We conclude that protein-energy restriction, starting early in life in the rat, severely impairs the release of counterregulatory hormones that defend against hypoglycaemia.
Acute Disease↗
Lack of deterioration of insulin action with aging in the GK rat: a contrasted adaptation as compared with nondiabetic rats.
One of the main characteristics of non-insulin-dependent diabetes mellitus (NIDDM) is an alteration of tissue insulin sensitivity, which is also observed during the aging process in the nondiabetic. In this study, we evaluated the influence of age on insulin resistance in a genetic lean model of NIDDM, the Goto-Kakisaki (GK) rat, using the euglycemic-hyperinsulinemic clamp technique at 2, 12, and 18 months of age. In GK rats, basal hyperglycemia (11 mmol/L) and insulinemia, glucose intolerance, and the specific failure of the insulin response to glucose apparent at 2 months of age remained stable until 18 months. Whatever the age, the insulin-suppressive effect on glucose production was significantly less in GK rats than in Wistar rats. The insulin effect on whole-body glucose utilization was decreased at 2 months (15.8 +/- 1.0 mg/min/kg v 23.5 +/- 2.0, P < .001) and was only mildly aggravated between 2 and 18 months (10.3 +/- 0.9 mg/min/kg, P < .05). By contrast, in Wistar control rats, basal insulinemia and the insulin response to glucose markedly increased between 2 and 18 months (2-month delta I v 18-month delta I, 1.4 +/- 0.1 mU/ml.min v 2.9 +/- 0.3, P < .001) and glucose tolerance remained normal. In 18-month-old Wistar rats, the insulin-stimulated glucose utilization rate (GUR) was found to be markedly decreased compared with that of 2-month-old Wistar rats (9.9 +/- 0.8 mg/min/kg v 23.5 +/- 2.0, P < .001), thus demonstrating an age-related decrease of insulin action. In conclusion, we find that there is no major alteration of insulin action due to aging in the GK rat, at variance with the pattern in nondiabetic rodents. It is speculated that such an adaptation in this lean model of NIDDM could be related to the limited capacity of these rats to expand their body weight with age, since it is recognized that body weight gain is largely responsible for the age-related impairment in peripheral insulin action in nondiabetic humans and nondiabetic animal models.
Aging↗
Insulin secretion in adult rats that had experienced different underfeeding patterns during their development.
Four populations of 70-day-old undernourished rats were studied. All rats were submitted to a 65% restriction of food intake as follows: 1) undernutrition of the mother from day 14 of gestation until birth only (fetal period, group F), 2) undernourished during fetal and suckling periods only (group FS), 3) undernourished from day 14 of gestation until day 70 of life (group FSA), and 4) undernourished from day 28 of life until day 70 (group A). Body weight, plasma glucose, glucose tolerance, basal plasma insulin, in vivo glucose-induced insulin secretion, pancreatic insulin content, and in vitro response of isolated islets to glucose and amino acids were investigated. Islet insulin content decreased in all four groups studied, and islet mass was found reduced in group FSA. Glucose tolerance was decreased in group A, and in vitro response of islets decreased in both groups FSA and A. We conclude that two different adaptive processes should be considered: the possible long-term consequences of impaired pancreas development due to inadequate perinatal nutrition and the functional alteration of insulin secretion to undernutrition in adulthood.
Aging↗
Impaired phosphoinositide metabolism in glucose-incompetent islets of neonatally streptozotocin-diabetic rats.
The effects of nutrient and neurotransmitter stimuli on insulin release, loss of phosphoinositides (PI), and production of inositol phosphates (InsP) were investigated in islets from neonatally streptozotocin-injected (nSTZ) rats. In islets from nSTZ rats, insulin secretory responses to 16.7 mM D-glucose and 10.0 mM D-glyceraldehyde were reduced compared with controls. Contents in phosphatidylinositol 4-monophosphate [PtdIns(4)P] and phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2], but not in phosphatidylinositol, were diminished. Glucose effects on breakdown of PtdIns(4)P and PtdIns(4,5)P2 and on total InsP accumulation were both reduced. D-Glucose was unable to increase the levels of both inositol trisphosphate isomers, Ins(1,3,4)P3 and Ins(1,4,5)P3. Glyceraldehyde also failed to promote InsP formation. By contrast, the ability of 1.0 mM carbachol or 300 nM cholecystokinin to stimulate insulin secretion and InsP generation was still observed. Thus a disturbed coupling between nutrient recognition and activation of phospholipase C, possibly together with a shortage of available polyphosphoinositides, could be responsible for the altered islet PI turnover in the nSTZ rats. It is proposed that such defects may contribute to the impairment of glucose-stimulated insulin secretion in this model of non-insulin-dependent diabetes mellitus.
Animals↗
Contrasted impact of maternal rat food restriction on the fetal endocrine pancreas.
The effects of food restriction of the mother (65% restriction of ad libitum food intake) on fetal and maternal insulin secretion and islet function were studied at 21 days gestation in three different rat populations: 1) undernourished from 0-7 days gestation, 2) undernourished from 7-14 days gestation, and 3) undernourished from 14-21 days gestation. The body weights of mothers were decreased in groups 2 and 3 vs. those in control fed pregnant animals, and no changes in basal parameters were found in any group. A glucose tolerance test in mothers from group 3 showed a mild intolerance to glucose and a decreased islet insulin content, although islet stimulation in vitro with glucose alone or plus arginine showed a normal insulin secretory response. Body weight was decreased in fetuses from the three groups (P < 0.01), and pancreas weight was reduced only in group 3. Insulinemia was increased in groups 2 and 3, and pancreatic insulin content increased only in group 3. However, fetuses from mothers of group 3 showed increased islet insulin content, increased response of insulin in vitro to glucose or glucose plus arginine, and hypertrophy of beta-cell mass. These results indicate, first, that the development of the fetal pancreas depends on a balanced maternal glucose homeostasis and, second, that adaptive maternal changes to undernutrition seem to induce alterations on the fetal endocrine pancreas, especially when food restriction is applied during the last week of gestation.
Animals↗
Glucagon-like peptide-1(7-36)-amide confers glucose sensitivity to previously glucose-incompetent beta-cells in diabetic rats: in vivo and in vitro studies.
The effects of glucagon-like peptide-1(7-36)-amide (GLP-1) on cAMP content and insulin release were studied in islets isolated from diabetic rats (n0-STZ model) which exhibited impaired glucose-induced insulin release. We first examined the possibility of re-activating the insulin response to glucose in the beta-cells of the diabetic rats using GLP-1 in vitro. In static incubation experiments, GLP-1 amplified cAMP accumulation (by 170%) and glucose-induced insulin release (by 140%) in the diabetic islets to the same extent as in control islets. Using a perifusion procedure, GLP-1 amplified the insulin response to 16.7 mM glucose by diabetic islets and generated a clear biphasic pattern of insulin release. The incremental insulin response to glucose in the presence of GLP-1, although lower than corresponding control values (1.56 +/- 0.37 and 4.53 +/- 0.60 pg/min per ng islet DNA in diabetic and control islets respectively), became similar to that of control islets exposed to 16.7 mM glucose alone (1.09 +/- 0.15 pg/min per ng islet DNA). Since in vitro GLP-1 was found to exert positive effects on the glucose competence of the residual beta-cells in the n0-STZ model. we investigated the therapeutic effect of in vivo GLP-1 administration on glucose tolerance and glucose-induced insulin release by n0-STZ rats. An infusion of GLP-1 (10 ng/min per kg; i.v.) in n0-STZ rats enhanced significantly (P < 0.01) basal plasma insulin levels, and, when combined with an i.v. glucose tolerance and insulin secretion test, it was found to improve (P < 0.05) glucose tolerance and the insulinogenic index, as compared with the respective values of these parameters before GLP-1 treatment.
Animals↗
Insulin administration enhances growth of the beta-cell mass in streptozotocin-treated newborn rats.
We have previously reported that the damage caused by streptozotocin (STZ) administration to the beta-cells in newborn rats was followed by spontaneous recovery from neonatal diabetes. Our present data indicate that STZ administration on the day of birth (day 1) reduced the total beta-cell mass on day 4 to only 10% of the normal value and that after such damage, 27% of the corresponding normal beta-cell mass was spontaneously regained on day 7. During days 4-7, the contribution of the predicted beta-cell growth (due to the replication of preexisting differentiated beta-cells) to the total beta-cell growth represented only 56%. Therefore, recruitment of new beta-cells from a precursor pool indeed represents a significant mechanism for beta-cell regeneration after STZ during this period of life. Here, we report for the first time that 1) insulin therapy from days 2 to 4 did not significantly influence the occurrence of beta-cell damage after STZ administration (total beta-cell mass on day 4 was reduced to 12% of the normal value) and 2) insulin therapy from days 2 to 6 did improve the otherwise spontaneous beta-cell regeneration, since on day 7 total beta-cell mass was 44% of the corresponding normal beta-cell mass. During days 4-7, the contribution of the predicted beta-cell growth to the total beta-cell growth represented only 32% in the insulin-treated STZ group. Finally the insulin-favored regeneration of the beta-cells reflects both an increased replication from differentiated beta-cells and an increased neogenesis from precursor/stem cells, with this last pathway being preferentially activated.
Animals↗
Decreased ATP-induced synthesis and Ca(2+)-stimulated degradation of polyphosphoinositides in pancreatic islets from neonatally streptozotocin-diabetic rats.
Phosphoinositide (PI) synthesis and hydrolysis were investigated in pancreatic islet homogenates from neonatal streptozotocin diabetic (n-STZ) and control rats. In the diabetics, ATP, in absence of Ca2+, failed to increase the amount of phosphatidylinositol 4-phosphate (PtdInsP) and phosphatidyl inositol 4, 5-bisphosphate (PtdInsP2) at variance with the pattern in controls. Also, the Ca(2+)-stimulated generation of inositol phosphates (InsP) was dramatically decreased, whether in the absence or presence of ATP. Moreover, phosphatidylinositol (PtdIns) kinase activity was reduced while PtdInsP kinase activity was not impaired. These data suggest that the suppressed formation of PtdInsP and subsequent PtdInsP2 synthesis, concomitantly with a decreased Ca(2+)-stimulated phospholipase C activity, may participate to the alteration of the PI pathway, the limitation of the InsP production, and finally the impairment of the insulin release in the n-STZ model of non-insulin-dependent diabetes.
1-Phosphatidylinositol 4-Kinase↗
Chromosomal mapping of genetic loci associated with non-insulin dependent diabetes in the GK rat.
Goto-Kakizaki (GK) rats are a well characterized model for non-insulin dependent diabetes mellitus (NIDDM). We have used a combination of physiological and genetic studies to identify quantitative trait loci (QTLs) responsible for the control of glucose homeostasis and insulin secretion in a F2 cohort bred from spontaneously diabetic GK rats. The genetic dissection of NIDDM allowed us to map up to six independently segregating loci predisposing to hyperglycaemia, glucose intolerance or altered insulin secretion, and a seventh locus implicated in body weight. QTLs implicated in glucose tolerance and adiposity map to the same region of rat chromosome 1, and may indicate the influence of a single locus. Our study demonstrates that distinct combinations of genetic loci are responsible for different physiological characteristics associated with the diabetic phenotype in the GK rat, and it constitutes an important step for directing the search for the genetic factors involved in human NIDDM.
Animals↗