Functional correlates of Na+,K+-ATPase in lean and obese (ob/ob) mice

Concentrations of Na+,K+-ATPase enzyme units are lower in skeletal muscle and liver of adult obese (ob/ob) mice than in their lean counterparts. The present studies were designed to provide information on functional correlates of Na+,K+-ATPase in ob/ob mice. Obese mice had lower potassium (K+) content in muscle and liver and higher sodium (Na+) content in muscle and liver and higher sodium (Na+) content in muscle than lean counterparts. The calculated intracellular Na+/K+ ratio in muscle of obese mice was approximately twice as high as in muscle of lean mice. Oxygen consumption was measured in mice maintained at 14 degrees, 25 degrees, or 33 degrees for 40 min and injected with 0.3 or 0.9 microgram ouabain per g body weight. Ouabain, a specific inhibitor of Na+,K+-ATPase, decreased oxygen consumption less in obese mice (12%--25%) than in lean mice (19%--38%). These results suggest that Na+ pump activity may be reduced in obese mice.     

Brown adipose tissue: contributions of nature and nurture to the obesity of an obese mutant mouse (ob/ob)

The aim of this investigation was to compare the contributions of the genotype of the brown adipose tissue (BAT) and of its environment to the obesity of the mutant mouse C57 BL/6J ob/ob. Pieces of interscapular BAT from lean or obese mice were transplanted to a site underneath the kidney capsule of recipient lean or obese mice. The grafts were left in place for 6 to 12 weeks and then examined by histological methods by electron microscopy to examine the ultrastructure of the mitochondria and by fluorescence histochemistry to examine the catecholaminergic innervation of the grafts. When lean BAT was grafted into obese mice, or when obese BAT was grafted into lean mice, kept at ambient temperatures, the characteristics of the donor BAT (i.e. lipid droplet size, mitochondrial ultrastructure and catecholaminergic innervation) transformed partially, but not completely, towards those of BAT in the host mouse. However, if lean mice containing obese BAT grafts were cold-acclimated at 4 degrees C or obese mice containing lean BAT grafts were warm-acclimated at 33 degrees C, the characteristics of the donor BAT transformed completely towards those of the BAT in the host mouse. This complete transformation occurred even if the host mice were returned to 23 degrees C after the period of temperature acclimation. Fluorescent histochemical observations indicated that the sympathetic innervation of BAT grafts was only indistinguishable from that of the lean or obese host BAT when the mice received a period of temperature acclimation (cold for lean mice; warm for obese mice). We conclude that BAT grafts from lean mice can assume the typical characteristics of BAT in obese hosts and that BAT grafts from obese mice can assume the typical characteristics of BAT in lean hosts provided that both the sympathetic innervation and the vascularization of the grafts is the same as in the host. Intrinsic properties of BAT in genetically obese mice are therefore unlikely to be of paramount importance in determining the obesity of the ob/ob mouse. Our results support the conclusions of other workers in implicating the low activity of the sympathetic innervation of BAT as being crucially important in causing the reduction of thermogenic activity.     

Effects of a warm environment on energy balance in obese (ob/ob) mice

Low rates of thermoregulatory heat production associated with low metabolic activity of brown adipose tissue, the primary site of thermoregulatory heat production, contribute substantially to the high efficiency of energy retention in obese (ob/ob) mice housed at 20 degrees C to 28 degrees C. To eliminate the need for thermoregulatory heat production lean and ob/ob mice were housed at 34.5 degrees C. At this temperature ob/ob mice still retained energy with a greater efficiency than lean littermates. Next, we investigated the possibility that the high efficiency of energy retention in ob/ob mice housed at 34.5 degrees C was related to depressed dietary-induced thermogenesis associated with low metabolic activity of brown adipose tissue. The sympathetic nervous system is a primary regulator of brown adipose tissue metabolism. Therefore, rates of norepinephrine (NE) turnover in brown adipose tissue, as an indicator of sympathetic nervous system stimulation of the tissue, were measured. Lean and ob/ob mice housed at 34.5 degrees C had equally low rates of NE turnover in their brown adipose tissue. Thus, the high efficiency of energy retention in ob/ob mice maintained at 34.5 degrees C is caused by factors other than low sympathetic stimulation of brown adipose tissue.     

An enzymatic defect in the obese (ob/ob) mouse: Loss of thyroid-induced sodium- and potassium-dependent adenosinetriphosphatase

Genetically obese (ob/ob) mice, mice that became obese after treatment with gold thioglucose, and lean animals were studied in the euthyroid state, after induction of hypothyroidism, and after treatment with triiodothyronine. The activity of glycerol 3-phosphate dehydrogenase (sn-glycerol-3-phosphate:(acceptor) oxidoreductase; EC 1.1.99.5] was reduced in the livers from hypothyroid animals and was increased by treatment with triiodothyronine in all groups. The activity of the ouabain-suppressible sodium- and potassium-dependent ATPase (ATP phosphohydrolase; EC 3.6.1.3) was increased by triiodothyronine and reduced by hypothyroidism in the lean and gold thioglucose-treated obese animals. In the obese (ob/ob) mice, on the other hand, treatment with triiodothyronine did not increase the activity of this enzyme, which remained at the level found in hypothyroid animals. This enzymatic activity was reduced in both liver and kidney. Adenylate cyclase [ATP pyrophosphate-lyase (cyclizing); EC 4.6.1.1] activity in liver membranes, however, was similar in all three groups of mice. This enzyme complex was activated by glucagon and was unaffected by treatment with thyroid hormones. The lack of a thyroid-dependent ouabain-suppressible (Na(+) + K(+))-ATPase in the tissues of the obese (ob/ob) mouse could explain most, if not all, of the abnormalities that have been described in this animal.     

Abnormal thyroid hormone deiodination in tissues of ob/ob and db/db obese mice

To explore the relationships between thyroid hormone economy and defective thermogenesis in two genetically obese mouse strains, ob/ob and db/db, we measured iodothyronine deiodination in tissue homogenates and the T4 and T3 content of selected tissues. In lean mice acclimated at room temperature, type II 5'-deiodination in brown adipose tissue (BAT) homogenates rose 14.5-fold between 30 and 180 min of cold exposure at 4 C. In ob/ob mice kept at room temperature, type II 5'-deiodination rates in BAT homogenates were 2.29 times those in lean controls, but acute exposure to 4 C for 3 h caused a much smaller rise in 5'-deiodination in the ob/ob mice than in lean controls. Administration of 1 microgram/g norepinephrine caused similar elevations in BAT 5'-deiodination in ob/ob and lean mice. db/db mice had a defect in cold-stimulated BAT 5'-deiodination similar to that in the ob/ob mice in two of three experiments. In the obese animals of both strains, but not the lean mice, the magnitude of the rise in BAT type II deiodination after cold exposure was much greater in December than in warm months. Cerebrocortical homogenates from ob/ob mice had about 33% higher type II 5'-deiodination rates than those from lean controls, but showed normal type III T3 5'-deiodination. ob/ob liver homogenates had about 33% lower type I 5'-deiodination rates than controls. db/db, but not ob/ob, mice had lower serum T4 concentrations than controls. Both ob/ob and db/db mice had lower hepatic T4 concentrations than lean controls. The ob/ob animals had no alteration in brain T4 concentrations. Neither ob/ob or db/db mice had altered serum T3 concentrations, ob/ob mice had subnormal hepatic, but not brain, mean T3 concentrations. db/db mice also had reduced mean heptic T3 concentration. These results suggest that the subnormal cold-induced rise in ob/ob and db/db BAT type II 5'-deiodination is unrelated to thyroid status, perhaps being partly caused by defective sympathetic stimulation. This impaired deiodination response might result in subnormal intracellular T3 in BAT, thereby contributing to impaired thermogenesis. In contrast, the elevated cerebrocortical and unstimulated BAT type II deiodination and the decreased hepatic type I deiodination in the ob/ob mice are probably due to intracellular thyroid hormone deficiency.     

Insulin secretion and the morphological and metabolic characteristics of pancreatic islets of hyperthyroid ob/ob mice.

Thyroxine treatment induced experimental hyperthyroidism in ob/ob mice, inhibited glucose-induced insulin secretion from the isolated perfused ob/ob mouse pancreas, and reduced total pancreas insulin content. In contrast, glucose-induced insulin release from incubated pancreatic islets and insulin content of pancreatic islets from ob/ob mice isolated by freehand microdissection were not reduced after thyroxine treatment when expressed per microgram dry islet. Histological examination of the ob/ob mouse pancreas revealed islets without degenerative lesions of islet cells. Granularity of beta cells was well preserved. The average number of pancreatic islets was unchanged. However, the beta cell area was significantly decreased in relation to the total pancreatic parenchyma after thyroxine treatment. This implies that insulin release and content per pancreatic islet was half of that of the controls. ATP content of islets was slightly reduced. Glucose oxidation and glucose utilization by islets from treated mice were slightly increased. Thyroxine treatment of the animals did not abolish the stimulation of 45Ca2+ uptake by glucose, but it did suppress the potentiating effect of fasting on the stimulatory effect of glucose on 45Ca2+ uptake. The metabolic characteristics of islets from experimentally hyperthyroid mice are those of all hyperthyroid tissues. The results provide no evidence for the view that the effects of thyroxine treatment may be due to disturbed metabolic function or energy deprivation of pancreatic islets. Inhibition of insulin secretion from the pancreas after thyroxine administration is apparently due to a reduction in pancreas insulin content and a diminished pancreatic islet volume. Reduced pancreatic islet volume represents most probably a reduction of individual islet cell volume.     

Alterations of brown adipose tissue in genetically obese (ob/ob) mice. II. Studies of beta-adrenergic receptors and fatty acid degradation

beta-Receptor number, norepinephrine-stimulated adenylate cyclase activity and lipolysis, octanoate-induced NAD(P) redox changes, and heat production were studied in brown adipose tissue (BAT) of lean and obese (ob/ob) mice. beta-Receptor number was increased 1.54-fold in purified brown adipocyte plasma membrane of ob/ob mice compared to that in lean controls. This increase was reversed by cold adaptation (5 C). Basal and norepinephrine-stimulated adenylate cyclase values were not different in the two groups. Norepinephrine stimulated lipolysis at 10 nM in BAT of lean mice, but only at 10 microM in BAT ob/ob mice. Octanoate produced an increase in the NAD(P) redox state in BAT of lean mice, but it did not modify the NAD(P) redox state in BAT of ob/ob mice. Concomitantly, octanoate increased heat production 3-fold in BAT of lean mice, but did not promote any significant increase in heat production in BAT of ob/ob mice. These two parameters were restored toward values observed in lean mice when the ob/ob mice were adapted to a cold environment. The data indicate that BAT of ob/ob mice exhibits three alterations; one at the level of the beta-receptor, one at the level of the lipolytic response to norepinephrine, and one at the level of fatty acid activation and/or beta-oxidation.     

The effect of limit feeding on thermogenesis and thermoregulation in genetically obese (ob/ob) mice during cold exposure

Thermogenesis and thermoregulation in ad-lib-fed and limit-fed lean (+/ob or +/+) and obese (ob/ob) mice during acute cold exposure were studied by measuring oxygen consumption and body temperature. No significant differences in oxygen consumption were found between the lean ad-lib, obese ad-lib- or obese limit-fed groups. The oxygen consumption of the lean-limit-fed group was decreased by 25-30 per cent compared with the other groups. The body temperature of the obese ad-lib-fed group fell at a rate of at least twice that of any other group. The weight, total cytochrome c oxidase activity and protein content of the brown adipose tissue (BAT) of the lean groups was similar, and there appeared to be little difference in cell size or fat content. The BAT of both obese groups showed a several-fold increase in weight, and a 50 per cent increase in total protein, compared with the lean groups. The limit-fed obese group showed a significant increase in cytochrome c oxidase activity compared with all other groups. The BAT cells of both obese groups were much enlarged and contained considerable amounts of fat. These observations indicate that the susceptibility of obese mice to hypothermia is not due to a reduced capacity for thermogenesis, but to a failure to conserve heat. Failure of thermoregulation in obese animals may be due to postural constraints that result in increased heat loss by radiation. The results are discussed in relation to the accredited role of BAT thermogenesis in rodents exposed to the cold.     

Acute and chronic regulation of ob mRNA levels by beta3-adrenoceptor agonists in obese Yellow KK mice.

The inhibitory effect of beta3-adrenoceptor agonists on the ob gene in brown adipose tissue (BAT) and white adipose tissue (WAT) is now well documented both in vivo in lean animals and in vitro, but the reported effects of beta3-adrenoceptor agonists on ob gene expression in obese animals remain controversial. We investigated whether ob gene expression in BAT and WAT is reduced by acute and chronic administrations of a beta3-adrenoceptor agonist, CL316,243 (CL). The ob gene mRNA levels in BAT, perimetric and inguinal WAT of obese Yellow KK mice were about 4-fold higher than those of lean controls. Acute exposure (6 h) to CL decreased ob gene mRNA levels in three fat depots in both animals. Chronic exposure (10 days) to CL also decreased ob gene mRNA levels in BAT, perimetric, and inguinal WAT in both animals. We concluded that acute and chronic regulation by a beta3-adrenoceptor agonist suppressed ob gene expression in obese Yellow KK mice and lean controls.     

Skeletal muscle accretion and turnover in lean and obese (ob/ob) mice.

Skeletal muscle growth in obese (ob/ob) and lean mice was estimated from changes in fat-free carcass weight during development. No differences were observed at 2 wk of age, but fat-free carcasses of obese mice weighed less than those of lean mice at 5 wk of age. Fat-free carcasses of the adult obese mice were only 78% as heavy as those of lean mice, although the obese mice weighed up to twice as much as the lean mice. Even though obese mice accumulated less muscle than lean mice, urinary creatinine output of the obese mice was equal to that of the lean mice. The amount of 3-methylhistidine (3MH) in the carcasses of the mice and urinary output of 3MH were used to calculate the fractional breakdown rate (FBR) of myofibrillar proteins. Accumulation of 3MH in the carcasses paralleled carcass weight gain; thus, obese mice accumulated less 3MH than lean mice. Urinary output of 3MH was measured from 5 to 37 wk of age and was as great in the obese as in the lean mice; consequently, the FBR of the myofibrillar proteins was faster in the obese than in the lean mice. The FBR averaged 8.5 ± 0.6%/day for obese mice and 5.0 ± 0.1%/day for lean mice. The rate of 3MH accumulation in the carcass and the FBR were subsequently utilized to calculate a fractional synthesis rate (FSR) for the myofibrillar proteins. The FSR was not reduced in obese mice. The results suggest that obese mice accumulate less muscle because the rate of muscle degradation is increased.     

Increased responsiveness to the hyperglycemic, hyperglucagonemic and hyperinsulinemic effects of circulating norepinephrine in ob/ob mice

OBJECTIVE: Several studies have implicated increased sympathetic tone as a contributing factor to the hyperglycemia and hyperglucagonemia of ob/ob mice. However, the responsiveness of plasma glucose, insulin and glucagon to circulating norepinephrine (NE) in ob/ob vs normal lean mice has never been described. Therefore, the present study investigated the effect of a 15 min intravenous NE infusion (1 pmol/min/g) on plasma glucose, insulin and glucagon in anesthetized lean, ob/ob, ob/ob-concurrent yohimbine (alpha(2) antagonist) treated, and ob/ob-chronically sympatholytic dopamine agonist treated (for 14 days prior to infusion) mice. In an effort to gain insight into a possible relation between norepinephrine, hyperglucagonemia and hyperinsulinemia in ob/ob mice, this study also examined the isolated islet responses to NE and glucagon in lean, ob/ob and ob/ob-sympatholytic dopamine agonist treated mice. RESULTS: Basal humoral values of glucose, insulin and glucagon were all elevated in ob/ob vs lean mice (by 63, 1900 and 63%, respectively, P<0.01). However, NE infusion further increased levels of glucose, insulin and glucagon in ob/ob (by 80, 90 and 60%, respectively, P<0.05) but not in lean mice (between group difference for all parameters P<0.05). Acute concurrent yohimbine treatment as well as chronic prior sympatholytic dopamine agonist treatment (bromocriptine plus SKF38393) simultaneously strongly aborgated or abolished all these humoral hypersensitivity responses to intravenous NE in ob/ob mice (P<0.05). Clamping the plasma glucose level in untreated ob/ob mice at a high level (30 mM) established by NE infusion did not significantly alter the plasma insulin level, suggesting that some other influence of NE was responsible for this insulin effect. Direct NE administration at 1 microM to islets from lean and ob/ob mice inhibited 15 mM glucose-stimulated insulin secretion in both groups, but at 0.1 microM it was inhibitory only in islets from ob/ob mice. However, glucagon (10 nM) increased 15 mM glucose-stimulated insulin secretion in ob/ob (by 170%, P<0.05) but not lean mice (between group difference P<0.05). CONCLUSION: These findings suggest that hypersensitivity to circulating NE may potentiate hyperglycemia and hyperglucagonemia in ob/ob mice, and the subsequent hyperglucagonemia coupled with increased islet beta-cell insulin secretory responsiveness to glucagon in ob/ob mice may support hyperinsulinemia, thus explaining the increased plasma insulin level response to intravenous NE in these animals. These findings further support a role for increased peripheral noradrenergic activities in the development and maintenance of the hyperglycemic, hyperglucagonemic and hyperinsulinemic state, characteristic of type 2 diabetes.     

Glycerol release in vitro from adipose tissue of obese (ob/ob) mice treated with thyroid hormones.

Glycerol release from adipose tissue of adult genetically obese (ob/ob) mice was greater than that from adipose tissue of lean litter-mates. However, a higher concentration of adrenaline was required to stimulate lipolysis in ob/ob adipose tissue. Since oxygen consumption and the uptake of radioactive iodine by the thyroid were reduced and the biological half-life of thyroidal radio-iodine was lengthened, it is suggested that ob/ob mice are hypothyroid. The poor sensitivity of ob/ob adipose tissue to low doses of adrenaline was improved by treatment with either tri-iodothyronine or thyroxine. The lipolytic responses to both dibutyryl 3':5'-cyclic adenosine monophosphate and aminophylline were enhanced in thyroid hormone-treated lean and ob/ob mice. A higher dose of tri-iodothyronine was required to enhance lipolysis in ob/ob mice than in lean controls. The possibility that hypothyroidism is responsible for the change in response of ob/ob adipose tissue to lipolytic hormones is discussed.     

The effects of exercise and food restriction on obesity and diabetes in young ob/ob mice

Hormone levels and body composition were examined in six-week-old C57BL/6J ob/ob mice following 25 d of limited caloric intake, voluntary exercise, or combined treatment. Pair-feeding obese mice to the daily intakes of lean mice reduced body weight gain, skeletal growth and lean body mass. Although weight gain was the same in the two phenotypes, ob/ob mice had fourfold higher rates of fat deposition. When exercise was combined with pair-feeding, skeletal and lean body growth were reduced even further and weight gain was now less than ad libitum-fed lean controls. Carcass fat accretion, however, continued to be two to three times greater. No single treatment reversed the hyperglycemia or elevated hormone production of obese mice, although slightly lower values of glucose, insulin, glucagon and corticosterone were associated with pair-feeding. When diet was combined with exercise, fasting glycemia and glucagonemia were reduced to equal the values of lean mice but insulin and corticosterone levels remained elevated. The present results show that dieted and exercised ob/ob mice continue to exhibit very high rates of fat deposition even though skeletal and lean growth are severely limited. Since fat accretion is maintained under these conditions, it appears that obese mice are not just storing excess calories as fat, but are actively regulating body fat content to levels about 30 percent higher than lean mice.     

Cold acclimation of obese (ob/ob) mice: effects of energy balance

Obese (ob/ob) and lean mice at 4 weeks of age were housed at 23 degrees C or 14 degrees C for 4 to 8 weeks to examine effects of acclimation to mild cold on energy balance. Energy intake of young lean mice increased by about 50% when housed at 14 degrees C, but energy intake of cold-acclimated obese mice increased by only 8%. Efficiency of energy retention (ratio of energy gain to energy intake) in obese mice declined from 22% +/- 1.2% at 23 degrees C to 10% +/- 1.8% after 4 weeks at 14 degrees C. Lean mice exhibited a less pronounced response to temperature; their efficiency of energy retention declined from 7% +/- 1.3% at 23 degrees C to 4% +/- 2.2% after 4 weeks at 14 degrees C. After 8 weeks of cold exposure, body weights and efficiency of energy retention became equal in obese and lean mice. Calculated heat production of cold-acclimated obese and lean mice was 40% higher than that of respective controls. Obese mice reacclimated to 23 degrees C after being kept for 4 weeks at 14 degrees C consumed the same amount of energy and were 16% more efficient than obese maintained at 23 degrees C; reacclimated lean mice consumed 12% more energy but were 53% less efficient than lean mice maintained at 23 degrees C. The results indicate that obese mice are able to increase heat production and markedly reduce their efficiency of energy retention when acclimated to mild cold but that they, unlike lean mice, rapidly revert to a high efficiency of energy retention after 4 weeks of reacclimation to 23 degrees C.     

Vitamin C supplementation decreases insulin glycation and improves glucose homeostasis in obese hyperglycemic (ob/ob) mice

The effects of dietary vitamin C supplementation on glucose homeostasis and insulin glycation were examined in adult lean and obese hyperglycemic (ob/ob) mice. In lean mice, supplementation of the drinking water with vitamin C (25 g/L) for 14 days did not affect food intake, fluid intake, glycated hemoglobin, plasma glucose, or plasma insulin concentrations. Total pancreatic insulin content and the percentage of glycated pancreatic insulin were also similar to control lean mice. In ob/ob mice, vitamin C supplementation caused significant reductions by 26% to 48% in food intake and fluid intake, glycated hemoglobin, plasma glucose, and insulin concentrations compared with untreated control ob/ob mice. The total insulin content and the extent of insulin glycation in the pancreas of ob/ob mice were also significantly decreased by 42% to 45% after vitamin C supplementation. This change was accompanied by a significant 80% decrease in the percentage of glycated insulin in the circulation of vitamin C-supplemented ob/ob mice. These data demonstrate that vitamin C supplementation can decrease insulin glycation and ameliorate aspects of the obesity-diabetes syndrome in ob/ob mice.     

Effects of streptozotocin on glucose metabolism, insulin response, and adiposity in ob/ob mice.

To determine whether hyperglycemia in the obese hyperglycemic (ob/ob) mouse is related to enhanced activity of the pancreatic beta cell, streptozotocin (175 mg/kg) was injected into lean and ob/ob mice at 8 wk of age. The influence of this injection upon glucose metabolism, adipose cellularity, pancreatic morphology and immunoreactive insulin (IRI) release from isolated pancreatic islets was measured. The plasma glucose levels before and after an oral glucose load were elevated in lean and decreased in ob/ob mice 2 wk after treatment with streptozotocin. By 5 wk after this treatment, a reduced pancreatic islet size, beta cell number and a decreased pancreatic islet IRI release were present in both lean and ob/ob mice. At this time, plasma glucose was still elevated in lean, but depressed in ob/ob mice and the insulin responsiveness in muscle and adipocytes was unchanged. Hyperglycemia abates in the ob/ob mouse as hypersecretion of insulin is diminished, but these observations may not be directly related, since streptozotocin affects key metabolic activities of the livers as well as the pancreatic beta cell. The progression of obesity and status of adipose cellularity are not directly related to hyper-insulinemia, since they are not altered following streptozotocin treatment.     

Pancreatic polypeptide and other hormones in pancreas of obese (ob/ob) mice

Summary The insulin, glucagon, somatostatin and pancreatic polypeptide content of acid-ethanol extracts of pancreas from lean and obese (ob/ob) mice of various ages was determined by radioimmunoassay. Rat and mouse pancreatic polypeptide react weakly with antibodies to avian, bovine and canine pancreatic polypeptide, but immunoassay for this peptide was possible using an antibody to the carboxyl-terminal hexapeptide of bovine pancreatic polypeptide and amino-terminal labelled bovine pancreatic polypeptide as tracer. The insulin, glucagon and pancreatic polypeptide content of pancreas from obese mice was greater than that of lean controls. The increase in insulin content uniformly involved ventral, dorsal and splenic lobes. Glucagon content was elevated primarily in the splenic lobe. Pancreatic polypeptide content was most significantly elevated in the splenic lobe where pancreatic polypeptide cells are infrequent in normal lean mice and this was accompanied by increased numbers of pancreatic polypeptide cells per islet in this lobe.     

Pancreatic islets of obese hyperglycemic mice (ob/ob).

The development of the obesity-associated hyperglycemic syndrome in ob/ob mice, genetically determined, was observed over time by a combined functional and structural study of pancreatic islets. Islet areas increased with advancing age in ob/ob mice from 2 times at 1 month to 30 times at 6 months of age the size of lean mouse islets. Islet areas apparently increased more than pancreatic insulin content in ob/ob mice. Glucose and insulin tolerance tests were performed to study in vivo responses to glucose and insulin, respectively, in 1-, 3-, and 6-month-old mice. With ob/ob mice, glucose tolerance tests revealed more elevation of plasma glucose than in lean mice, the lean mice revealing more elevated plasma insulin than the obese mice. In insulin tolerance tests, lean mice presented marked hypoglycemia, whereas ob/ob mice revealed slightly higher plasma glucose at 1 month of age but three to four times higher amounts than that of lean mice at 6 months of age. Thus, increasing insulin resistance in ob/ob mice older than 3 months is associated with progressively increasing islet area, which contains proportionally less insulin than that of lean mouse pancreas. The data suggest that insulin resistance in ob/ob mice progressively develops up to 6 months of age and that marked islet hyperplasia is likely in response to sustained hyperglycemia, leading to hyperinsulinemia and eventual marked obesity.     

Age dependent effects of oxytetracycline in ob/ob mice

Summary The effects of daily oxytetracycline injections on the metabolism and growth of prepubertal, 10 and 24 week old ob/ob and lean mice were examined. Drug treatment (100 mg/kg) between 17 and 28 days of age similarly reduced body weight gain and skeletal growth of both ob/ob and lean mice, but had little effect on obesity, serum glucose, or IRI levels. In older obese mice, 6 days of oxytetracycline led to substantial reductions of serum glucose, serum IRI, and obesity when compared with either ad libitum fed obese mice or food-restricted ob/ob mice serving as weight loss controls. In lean mice oxytetracycline also significantly reduced body weight and serum glucose, while no effects on obesity or IRI levels were noted. The present data show that oxytetracycline reduces the severity of many of the diabetic symptoms of ob/ob mice, but that only the ameliorative effects on obesity and hyperinsulinaemia appear to be specific to the ob/ob genotype.     

Adrenalectomy reduces but does not reverse obesity in ob/ob mice

Bilateral adrenalectomy (ADX) of 4-month-old ob/ob mice led to reduced rates of body weight gain, a complete cessation of fat deposition and increased percentage carcass protein and ash during a 2-month observation period after surgery. However, ADX obese mice were still heavier and had more body fat and lower concentrations of carcass protein and ash than intact sex-matched littermate lean mice at the end of the experiment. When adrenalectomy was performed in younger obese mice before the syndrome was fully expressed (23 +/- 2 days of age), body weight gain was reduced by 40 per cent and fat deposition by 50 per cent during the next 3.5 months, but each was still greater than that of littermate lean mice. Despite the lower rate of weight gain after adrenalectomy, the skeletal and lean body growth of the early ADX obese mice equalled that of both obese and lean mice fed ad libitum. When the carcass composition of early ADX obese mice was compared with that of intact obese mice which were calorically restricted to the same rate of body weight gain, the ADX group had significantly less carcass fat (28 per cent) and more protein (50 per cent) and ash (20 per cent) than the dieted obese mice. In both experiments adrenalectomy led to reduced circulating immunoreactive insulin levels, although hyperinsulinemia persisted. The present results show that adrenalectomy is an effective tool for ameliorating the severity of many aspects of the ob/ob syndrome, particularly when compared with caloric restriction, but the procedure does not entirely reverse the deranged metabolism or abnormal carcass composition of these mice.