Biomedical subjects

G L Dohm

Publications and source records attributed to G L Dohm.

At least 37 records · Page 2Linked to original sources

PubMed1998

A new paradigm for type 2 diabetes mellitus: could it be a disease of the foregut?

SUMMARY BACKGROUND DATA: We previously reported, in a study of 608 patients, that the gastric bypass operation (GB) controls type 2 diabetes mellitus in the morbidly obese patient more effectively than any medical therapy. Further, we showed for the first time that it was possible to reduce the mortality from diabetes; GB reduced the chance of dying from 4.5% per year to 1% per year. This control of diabetes has been ascribed to the weight loss induced by the operation. These studies, in weight-stable women, were designed to determine whether weight loss was really the important factor. METHODS: Fasting plasma insulin, fasting plasma glucose, minimal model-derived insulin sensitivity and leptin levels were measured in carefully matched cohorts: six women who had undergone GB and had been stable at their lowered weight 24 to 30 months after surgery versus a control group of six women who did not undergo surgery and were similarly weight-stable. The two groups were matched in age, percentage of fat, body mass index, waist circumference, and aerobic capacity. RESULTS: Even though the two groups of patients were closely matched in weight, age, percentage of fat, and even aerobic capacity, and with both groups maintaining stable weights, the surgical group demonstrated significantly lower levels of serum leptin, fasting plasma insulin, and fasting plasma glucose compared to the control group. Similarly, minimal model-derived insulin sensitivity was significantly higher in the surgical group. Finally, self-reported food intake was significantly lower in the surgical group. CONCLUSIONS: Weight loss is not the reason why GB controls diabetes mellitus. Instead, bypassing the foregut and reducing food intake produce the profound long-term alterations in glucose metabolism and insulin action. These findings suggest that our current paradigms of type 2 diabetes mellitus deserve review. The critical lesion may lie in abnormal signals from the gut.

Adult↗

PubMed1998

Regulation of glucose transporters GLUT-4 and GLUT-1 gene transcription in denervated skeletal muscle.

Because GLUT-4 expression is decreased whereas GLUT-1 expression is increased in denervated skeletal muscle, we examined the effects of denervation on GLUT-4 and GLUT-1 gene transcription. The right hindlimb skeletal muscle of male transgenic mice containing sequential truncations (2,400, 1,639, 1,154, and 730 bp) of the human GLUT-4 promoter linked to the chloramphenacol acyl transferase (CAT) gene was denervated, and the contralateral hindlimb was sham operated. RNase protection analysis revealed that after 72 h denervation decreased CAT mRNA and GLUT-4 mRNA levels 64-85%, respectively (P < 0.05), in the gastrocnemius muscles. In contrast, denervation of the right hindlimb of male rats increased GLUT-1 gene transcription and GLUT-1 mRNA levels by 94 and 213%, respectively (P < 0.05). In conclusion, GLUT-4 transcription is decreased but GLUT-1 transcription is increased in denervated skeletal muscle, suggesting that the effects of denervation on GLUT-4 and GLUT-1 expression are, in part, transcriptionally mediated. Furthermore, these data indicate that a DNA sequence regulated by denervation is located within 730 bp of the 5'-flanking promoter region of the human GLUT-4 gene.

Animals↗

PubMed1998

Membrane glycoprotein PC-1 and insulin resistance.

Peripheral resistance to insulin is a major component of non-insulin dependent diabetes mellitus. Defects in insulin receptor tyrosine kinase activity have been demonstrated in several tissues from insulin resistant subjects, but mutations in the insulin receptor gene occur in only a small fraction of cases. Therefore, other molecules that are capable of modulating the function of the insulin receptor are likely candidates in the search for the cellular mechanisms of insulin resistance. We have isolated an inhibitor of insulin receptor tyrosine kinase activity from cultured fibroblasts of an insulin resistant NIDDM patient and identified it as membrane glycoprotein PC-1. Subsequently we have demonstrated that expression of PC-1 is elevated in fibroblasts from other insulin resistant subjects, both with and without NIDDM. Studies in muscle, the primary site for insulin-mediated glucose disposal, have shown that the levels of PC-1 in this tissue are inversely correlated to insulin action both in vivo and in vitro. Transfection of PC-1 into cultured cells has confirmed that overexpression of PC-1 can produce impairments in insulin receptor tyrosine kinase activity and the subsequent cellular responses to insulin. Preliminary data suggests a direct interaction between PC-1 and the insulin receptor. However, the mechanisms whereby PC-1 inhibits insulin receptor signaling remain to be determined.

Animals↗

PubMed1997

Vanadate stimulation of 2-deoxyglucose transport is not mediated by PI 3-kinase in human skeletal muscle.

Glucose transport in mammalian skeletal muscle is stimulated by insulin, hypoxia and tyrosine protein phosphatase inhibitors such as vanadate. However, it is unknown whether the vanadate signaling mechanism shares a common or separate pathway with insulin or hypoxia. Therefore, experiments were conducted on incubated human muscle strips to compare the effects of vanadate with insulin and hypoxia stimulated 2-deoxyglucose transport (2-DOG). We also used the phosphatidylinositol 3-kinase (PI 3-kinase) inhibitor wortmannin to examine whether PI 3-kinase is a common step by which each stimulate glucose transport. Results demonstrate that whereas the effects of vanadate and hypoxia were additive with insulin stimulated glucose transport, the effect of vanadate plus hypoxia was not. In addition, wortmannin significantly (P < 0.05) reduced insulin but not vanadate or hypoxia stimulated 2-DOG transport. Moreover, PI 3-kinase activity was significantly elevated (P < 0.05) in the presence of insulin but not vanadate. In conclusion, these data suggest that vanadate and hypoxia stimulate glucose transport via a similar signaling pathway which is distinct from insulin and that the vanadate signaling pathway is not mediated by PI 3-kinase in human skeletal muscle.

Analysis of Variance↗

PubMed1997

Transcriptional regulation of hexokinase II in denervated rat skeletal muscle.

Hexokinase II protein is augmented in denervated skeletal muscle; therefore, we determined if hexokinase II gene transcription rates and mRNA levels are increased with denervation. The right hindlimb skeletal muscles of male rats were denervated while the left hindlimbs were sham operated. Seventy-two h following surgery, rats were sacrificed and the gastrocnemius and soleus muscles were harvested for nuclear and RNA isolation. Nuclear run-on and ribonuclease protection analyses indicated that denervation increased hexokinase II transcription rates and mRNA levels 42% and 88%, respectively (p < 0.05). Total hexokinase activity rose 23% in denervated gastrocnemius muscle. In conclusion, the increase in hexokinase II gene transcription and mRNA may account for the increase in hexokinase II protein and the subsequent rise in total hexokinase activity in denervated rat skeletal muscle.

Animals↗

PubMed1997

Treadmill running increases phosphatidylinostol 3-kinase activity in rat skeletal muscle.

Exercise has been shown to increase insulin-stimulated skeletal muscle glucose transport. Activation of phosphatidylinostol 3-kinase (PI 3-kinase) is required for insulin to stimulate glucose transport. The present study was designed to investigate whether treadmill running (60 min, 8% grade, 30 m/min) augments insulin-stimulated activation of PI 3-kinase. Insulin dramatically increased insulin receptor substrate-1 (IRS-1) tyrosine phosphorylation (p < 0.05). Treadmill running did not induce IRS-1 tyrosine phosphorylation, not did it alter insulin-stimulated IRS-1 tyrosine phosphorylation. Insulin increased PI 3-kinase activity by 3.0-fold (over basal activity) in white muscle and 5.2-fold in red muscle (p < 0.05). Exercise did not alter basal PI 3-kinase activity in either white or red muscle. However, in response to exercise, insulin-stimulated PI 3-kinase activity was significantly increased in both muscle fibers (p < 0.05). These results suggest that increased insulin responsiveness induced by exercise may be due, in part, to enhanced insulin-stimulated PI 3-kinase activity.

Animals↗

PubMed1997

Alterations in skeletal muscle protein-tyrosine phosphatase activity and expression in insulin-resistant human obesity and diabetes.

Obese human subjects have increased protein-tyrosine phosphatase (PTPase) activity in adipose tissue that can dephosphorylate and inactivate the insulin receptor kinase. To extend these findings to skeletal muscle, we measured PTPase activity in the skeletal muscle particulate fraction and cytosol from a series of lean controls, insulin-resistant obese (body mass index > 30) nondiabetic subjects, and obese individuals with non-insulin-dependent diabetes. PTPase activities in subcellular fractions from the nondiabetic obese subjects were increased to 140-170% of the level in lean controls (P < 0.05). In contrast, PTPase activity in both fractions from the obese subjects with non-insulin-dependent diabetes was significantly decreased to 39% of the level in controls (P < 0.05). By immunoblot analysis, leukocyte antigen related (LAR) and protein-tyrosine phosphatase 1B had the greatest increase (threefold) in the particulate fraction from obese, nondiabetic subjects, and immunodepletion of this fraction using an affinity-purified antibody directed at the cytoplasmic domain of leukocyte antigen related normalized the PTPase activity when compared to the activity from control subjects. These findings provide further support for negative regulation of insulin action by specific PTPases in the pathogenesis of insulin resistance in human obesity, while other regulatory mechanisms may be operative in the diabetic state.

Adult↗

PubMed1997

Mechanisms by which insulin and muscle contraction stimulate glucose transport.

Insulin binding to its receptor activates a tyrosine kinase that initiates a cascade of signaling events, the initial step being the tyrosine phosphorylation of insulin receptor substrate 1 (IRS-1). Subsequent IRS-1 association and activation of phosphatidylinositiol 3-kinase (PI 3-kinase) is believed to be involved in the events leading to the translocation of glucose transporters (GLUT4) to the plasma membrane resulting in uptake of glucose into the cell. Muscle contractions increase insulin sensitivity, but also stimulate muscle glucose uptake independent of insulin. The contraction signaling pathway is distinct from the insulin pathway because the effect of insulin and contractions on glucose uptake are additive, and contractions do not increase insulin receptor kinase or PI 3-kinase activity. In contrast, studies indicating that contractions cause the translocation of GLUT4 and that both contractions and insulin-stimulated glucose transport can be blocked by calcium channel blockers suggest that the two pathways may converge. However, the possibility that two distinct GLUT4 pools may be targeted, one by insulin the other by contractions, indicates that additional research is needed to better define the mechanisms by which glucose transport is stimulated in muscle.

Animals↗

PubMed1997

Eccentric contractions decrease glucose transporter transcription rate, mRNA, and protein in skeletal muscle.

We have recently shown that eccentric contractions (ECs; forced lengthening of active muscle) elicit a delayed decrease in glucose transporter (GLUT-4) protein content in rat skeletal muscle and a decrease in subsequent contraction-stimulated glucose transport. Here, we investigate whether this decrease in total GLUT-4 protein after prior ECs is due to changes in GLUT-4 gene transcription rate and GLUT-4 mRNA level. Furthermore, the effect of prior ECs on sarcolemmal GLUT-4 protein content in plasma membrane (PM) vesicles isolated from contraction-stimulated muscle was determined. Rat gastrocnemius muscle was electrically stimulated for ECs, and the contralateral muscle served, as unstimulated control (UC). Two days later, the total GLUT-4 protein content was decreased by 50% (P < 0.05) and 32% (P < 0.05) in the white and red gastrocnemius muscle, respectively. Furthermore, the GLUT-4 mRNA concentration was decreased by 41% (P < 0.05) in both the white and red gastrocnemius muscle. Moreover, the GLUT-4 transcription rate, determined by nuclear run-on analysis, was decreased by 75% (P < 0.05) in mixed EC gastrocnemius muscle compared with UC muscle. PM vesicles were isolated from EC and UC muscle after 15 min of isometric contractions. The PM GLUT-4 protein content was reduced by 51% (P < 0.05) in EC muscle compared with UC muscle. In conclusion, 2 days after ECs, the GLUT-4 transcription rate, GLUT-4 mRNA, and GLUT-4 protein content were decreased in rat skeletal muscle. Moreover, the PM GLUT-4 protein content in contraction-stimulated muscle was decreased. We suggest that eccentric muscle contractions decrease muscle GLUT-4 transcription rate, resulting in a lower GLUT-4 protein content, which in turn decreases the number of GLUT-4 transporters translocated to the sarcolemma, ultimately leading to decreased contraction-induced muscle glucose transport.

Animals↗

PubMed1997

Regulation of glucose transporter GLUT-4 and hexokinase II gene transcription by insulin and epinephrine.

Transport of glucose across the plasma membrane by GLUT-4 and subsequent phosphorylation of glucose by hexokinase II (HKII) constitute the first two steps of glucose utilization in skeletal muscle. This study was undertaken to determine whether epinephrine and/or insulin regulates in vivo GLUT-4 and HKII gene transcription in rat skeletal muscle. In the first experiment, adrenodemedullated male rats were fasted 24 h and killed in the control condition or after being infused for 1.5 h with epinephrine (30 microg/ml at 1.68 ml/h). In the second experiment, male rats were fasted 24 h and killed after being infused for 2.5 h at 1.68 ml/h with saline or glucose (625 mg/ml) or insulin (39.9 microg/ml) plus glucose (625 mg/ml). Nuclei were isolated from pooled quadriceps, tibialis anterior, and gastrocnemius muscles. Transcriptional run-on analysis indicated that epinephrine infusion decreased GLUT-4 and increased HKII transcription compared with fasted controls. Both glucose and insulin plus glucose infusion induced increases in GLUT-4 and HKII transcription of twofold and three- to fourfold, respectively, compared with saline-infused rats. In conclusion, epinephrine and insulin may regulate GLUT-4 and HKII genes at the level of transcription in rat skeletal muscle.

Adrenal Medulla↗

PubMed1997

Insulin activation of phosphatidylinositol 3-kinase in human skeletal muscle in vivo.

The purpose of this investigation was to determine whether insulin-stimulated phosphatidylinositol 3-kinase (PI3-kinase) activity is detectable in needle biopsies of human skeletal muscle. Sixteen healthy nonobese males matched for age, percent fat, fasting insulin, and fasting glucose participated in one of two experimental protocols. During an intravenous glucose tolerance test (IVGTT) protocol, insulin-stimulated PI3-kinase activity was determined from percutaneous needle biopsies at 2, 5, and 15 min post-insulin administration (0.025 U/kg). In the second group, a 2-h, 100 mU . m-2 . min-1 euglycemic hyperinsulinemic clamp was performed, and biopsies were obtained at 15, 60, and 120 min after insulin infusion was begun. Insulin stimulated PI3-kinase activity by 1.6 +/- 0.2-, 2. 2 +/- 0.3-, and 2.2 +/- 0.4-fold at 2, 5, and 15 min, respectively, during the IVGTT. During the clamp protocol, PI3-kinase was elevated by 5.3 +/- 1.3-, 8.0 +/- 2.6-, and 2.7 +/- 1.4-fold above basal at 15, 60, and 120 min, respectively. Insulin-stimulated PI3-kinase activity at 15 min post-insulin administration was significantly greater during the clamp protocol vs. the IVGTT (P < 0.05). These observations suggest that insulin-stimulated PI3-kinase activity is detectable in needle biopsies of human skeletal muscle, and furthermore, that the euglycemic, hyperinsulinemic clamp protocol may be a useful tool to assess insulin signaling in vivo.

Adult↗

PubMed1997

Effects of thyroid hormone on GLUT4 glucose transporter gene expression and NIDDM in rats.

Previous studies have shown that T3 coordinately stimulates GLUT4-glucose transporter messenger RNA (mRNA) and protein expression in mixed fiber-type skeletal muscle of the rat and produces a concomitant elevation in basal (noninsulin mediated) glucose uptake. The aim of the present study was to 1) determine the precise mechanism(s) for the T3-induced expression of GLUT4 in skeletal muscle, and 2) investigate the potential benefits of T3 on noninsulin dependent diabetes mellitus (NIDDM). Ten daily ip injections of T3 (100 micrograms/100 g BW) administered to hypothyroid male Sprague-Dawley rats, increased both GLUT4 mRNA and transcription approximately 70% (P < 0.05) in mixed fiber-type hindlimb skeletal muscle. Transcriptional induction was subsequently defined to be restricted to red (oxidative) muscle fibers (2.5-fold; P < 0.05), whereas GLUT4 protein was increased in both red and white (glycolytic) skeletal muscle. GLUT4 mRNA and protein expression were similarly inducible in the skeletal muscle of insulin-resistant Zucker rats. More importantly, T3 treatment totally ameliorated hyperinsulinemia in obese animals (P < 0.001), although their moderately elevated plasma glucose levels were not significantly altered. In conclusion, regulation of GLUT4 expression by T3 was shown to lie at the transcriptional level in red skeletal muscle, whereas in white muscle fiber types, it appears to operate via an alternative posttranscriptional mechanism. These data also support the potential of hormonally inducing glucose transporter expression in insulin-resistant muscle. However, high levels of T3 are associated with a number of adverse side-effects, in particular the stimulation of hepatic gluconeogenesis. Nevertheless, future studies may demonstrate, e.g. subthyrotoxic levels, to be similarly effective but without side effects, and thus perhaps find a clinical application in reducing both hyperinsulinemia and hyperglycemia in NIDDM.

Animals↗

PubMed1997

Characterization of a low affinity thyroid hormone receptor binding site within the rat GLUT4 gene promoter.

Previous studies have demonstrated that thyroid hormone (T3) stimulates insulin-responsive glucose transporter (GLUT4) transcription and protein expression in rat skeletal muscle. The aim of the present study was to define a putative thyroid hormone response element (TRE) within the rat GLUT4 promoter and thus perhaps determine whether T3 acts directly to augment skeletal muscle GLUT4 transcription. To this end, electrophoretic mobility shift analyses were performed to analyze thyroid hormone receptor (TR) binding to a previously characterized 281-bp T3-responsive region of the rat GLUT4 promoter. Indeed, within this region, a TR-binding site of the standard DR + 4 TRE variety was located between bases -457/ -426 and was shown to posses a specific affinity for in vitro translated TRs. Interestingly, however, the GLUT4 TR-binding site demonstrated a significantly lower affinity compared to a consensus DR + 4 TRE, and only bound TRs appreciatively in the form of high affinity heterodimers, in this case with the cis-retinoic acid receptor. In conclusion, these data demonstrated the presence of a specific TR-binding site within a T3-responsive region of the rat GLUT4 promoter and thus support the supposition that thyroid hormone acts directly to stimulate GLUT4 transcription in rat skeletal muscle. Moreover, characterization of a novel TR-binding site with low affinity suggests an additional mechanism by which the intrinsic activity and responsiveness of thyroid hormone regulated genes may be modulated.

Animals↗

PubMed1997

Leptin directly alters lipid partitioning in skeletal muscle.

Leptin, an adipocyte-derived hormone that directly regulates both adiposity and energy homeostasis, decreases food intake and appears to partition metabolic fuels toward utilization and away from storage. Because skeletal muscle expresses the leptin receptor and plays a major role in determining energy metabolism, we studied leptin's effects on glucose and fatty acid (FA) metabolism in isolated mouse soleus and extensor digitorum longus (EDL) muscles. One muscle from each animal served as a basal control. The contralateral muscle was treated with insulin (10 mU/ml), leptin (0.01-10 microg/ml), or insulin plus leptin, and incorporation of [14C]glucose or [14C]oleate into CO2 and into either glycogen or triacylglycerol (TAG) was determined. Leptin increased soleus muscle FA oxidation by 42% (P < 0.001) and decreased incorporation of FA into TAG by 35% (P < 0.01) in a dose-dependent manner. In contrast, insulin decreased soleus muscle FA oxidation by 40% (P < 0.001) and increased incorporation into TAG by 70% (P < 0.001). When both hormones were present, leptin attenuated both the antioxidative and the lipogenic effects of insulin by 50%. Less pronounced hormone effects were observed in EDL muscle. Leptin did not alter insulin-stimulated muscle glucose metabolism. These data demonstrate that leptin has direct and acute effects on skeletal muscle.

Animals↗

PubMed1996

The effect of exercise on ob gene expression.

Expression of the ob gene is subject to nutritional as well as hormonal regulation to control fat storage. In the present study we investigated the effect of acute exercise and long-term exercise training on ob mRNA levels in rat adipose tissue. Northern blot analysis showed that a single bout of exercise significantly decreased ob mRNA levels approximately 30% immediately and 3 hr after exercise. After 4 weeks of exercise training the ob mRNA level was significantly decreased approximately 48% at 2 hr after the last training session compared to the control group. These data demonstrated that exercise has a transient effect on ob gene expression and suggest that regulation of ob gene expression may help control energy balance following exercise.

Adipose Tissue↗

PubMed1996

Differential expression of ob mRNA in rat adipose tissues in response to insulin.

The recently cloned obese (ob) gene encodes a fat-specific mRNA and a protein which is believed to regulate satiety and metabolic rate so as to control fat storage. In the present study we investigated rat ob mRNA levels in different fat depots and the effects of insulin infusion on ob mRNA expression. Northern blot analysis showed the abdominal fat (epididymal and perirenal fat pads) had higher ob mRNA levels than subcutaneous fat. After a 2.5-hr infusion of insulin into fasted rats, ob mRNA levels were increased by 3.0 and 1.9-fold in epididymal and perirenal fat pads, respectively. However, no change of ob mRNA level was observed in the subcutaneous fat depot. These data demonstrate adipose depot site-specific expression of ob mRNA and support the hypothesis that insulin plays an important role in the expression of ob mRNA.

Adipose Tissue↗

PubMed1996

Effect of reduced training and training cessation on insulin action and muscle GLUT-4.

This study examined the impact of a 50% reduction in training frequency or training cessation on insulin action and muscle GLUT-4 protein concentration. Middle-aged individuals were tested before and after 12 wk of exercise training (4 days/wk, 40-45 min/day). Subjects then either maintained training (n = 9), reduced training frequency by 50% (n = 11), or stopped exercising (n = 10) for the ensuing 2 wk. GLUT-4 protein concentration and insulin action (insulin sensitivity index, as determined by the minimal model) increased (P < or = 0.05) by an average of 1.6- and 1.9-fold, respectively, with the 12 wk of training. Insulin action and GLUT-4 did not increase further with the additional 2 wk of training in the maintained training group. Similarly, insulin sensitivity index and GLUT-4 concentration remained at trained levels when training frequency was reduced by 50% for 2 wk. GLUT-4 concentration and insulin action, however, were not different from sedentary values after 14 days of training cessation. These findings indicate that a 14-day 50% reduction in exercise frequency maintains the improvements in GLUT-4 protein concentration and insulin action gained with endurance training in moderately trained middle-aged adults; in contrast, these adaptations are largely lost with training cessation.

Adult↗

PubMed1996

Skeletal muscle content of membrane glycoprotein PC-1 in obesity. Relationship to muscle glucose transport.

Membrane glycoprotein PC-1, an inhibitor of insulin signaling, produces insulin resistance when overexpressed in cells transfected with PC-1 cDNA. In the present study, we determined whether PC-1 plays a role in the insulin resistance of skeletal muscle in obesity. Rectus abdominus muscle biopsies were taken from patients undergoing elective surgery. Subjects included both NIDDM patients (n = 14) and nondiabetic patients (n = 34) across a wide range of BMI values (19.5-90.1). Insulin-stimulated glucose transport was measured in incubated muscle strips, and PC-1 content, enzymatic activity, and insulin receptor content were measured in solubilized muscle extracts. Increasing BMI correlated with both an increase in the content of PC-1 in muscle (r = 0.55, P < 0.001) and a decrease in insulin stimulation of muscle glucose transport (r = -0.58, P = 0.008). NIDDM had no effect on either PC-1 content or glucose transport for any given level of obesity. Insulin stimulation of muscle glucose transport was negatively related to muscle PC-1 content (r = -0.68, P = 0.001) and positively related to insulin receptor content (r = 0.60, P = 0.005). Multivariate analysis indicated that both skeletal muscle PC-1 content and insulin receptor content, but not BMI, were independent predictors of insulin-stimulated glucose transport. Muscle PC-1 content accounted for 42% and insulin receptor content for 17% of the variance in glucose transport values. These studies raise the possibility that increased expression of PC-1 and a decreased insulin receptor content in skeletal muscle may be involved in the insulin resistance of obesity.

Adult↗