Permanent hexose transport upregulation in a respiration-deficient human fibroblast cell strain

Andrejchyshyn, Susan; Continelli, Lia; Germinario, Ralph J.

doi:10.1152/ajpcell.1991.261.6.c973

Cited by 15 publications

(9 citation statements)

References 28 publications

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“…The lysate values of both cell types was <1 unit of the 5′‐nucleotidase activity (data not shown). This indicates that the increased whole cell GLUT 1 transporter content observed earlier (7) resides in the plasma membrane (i.e., the 5′‐nucleotidase–enriched fraction) and that the elevation in 2‐DG transport is due to the membrane‐located GLUT 1 transporter content.…”

Section: Resultsmentioning

confidence: 66%

“…Furthermore, our studies with the G14 mutant cell line derived from Chinese hamster lung fibroblasts (also NADH CoQ reductase − ) (7–11) produced the same enzyme defect and also exhibited increased sugar transport. Since early data indicated that sugar transport was increased in response to the mitochondrial defect, we sought to further characterize the mechanism(s) involved (7, 9, 11). In our continuing studies on the mechanism(s) involved in insulin resistance, elucidation of routes that cells employ in the control of glucose transport should shed some light on this problem.…”

mentioning

confidence: 59%

“…Details of the experimental protocols can be found elsewhere (7, 10). Briefly, cells were plated at 10 4 cells/cm 2 on plastic Petri dishes (35 mm diameter, Falcon, BD Biosciences, Oakville, Ontario) and grown to confluence.…”

Section: Methodsmentioning

confidence: 99%

“…Prior to electrophoresis, samples were solubilized in 2% sodium dodecyl sulfate (SDS) (Pierce Chemical Co., Rockford, IL), 10% glycerol, 1 m M EDTA, 0.002% bromphenol blue, and 100 m M TrisHCl (pH 6.8). Electrophoresis was performed as described previously (6, 7, 14). Purified human erythrocyte glucose transporter was prepared as described by others (17) and used as a control GLUT 1 in Western analysis.…”

Section: Methodsmentioning

confidence: 99%

“…We have reported earlier that the WG750 respiration‐deficient (i.e., NADH CoQ reductase − ) human fibroblast cell line exhibited increased whole cell levels of GT (7) and sugar transport. Furthermore, our studies with the G14 mutant cell line derived from Chinese hamster lung fibroblasts (also NADH CoQ reductase − ) (7–11) produced the same enzyme defect and also exhibited increased sugar transport. Since early data indicated that sugar transport was increased in response to the mitochondrial defect, we sought to further characterize the mechanism(s) involved (7, 9, 11).…”

mentioning

confidence: 97%

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Sugar Transport Regulation: Comparative Characterization of the Effect of NADH CoQ Reductase Deficiency in Two Cell Culture Systems

Germinario

Continelli

Pratt

2000

Proc Soc Exp Biol Med

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Abstract. In this report, we have characterized the upregulation of glucose transport in two different respiration‐deficient fibroblast cell cultures. We have demonstrated that glucose transport increases in respiration‐deficient cells as measured by 2 deoxy D‐glucose transport and is readily observed in both the WG750 human and G14 Chinese hamster fibroblast respiration‐deficient cell lines when compared with the MCH55 normal human and V79 parental Chinese hamster cell lines, respectively. Using subcellular fractionation techniques, the GLUT 1 glucose transporter was found located predominantly in the plasma membrane–enriched fraction of the human and hamster cell lines. In human cells, the expression of the GLUT 1 glucose transporter was elevated three‐fold in the plasma membrane–enriched fraction of the WG750 respiration‐deficient mutant cells. In the Chinese hamster cell lines, the respiration‐deficient G14 cells exhibited no such GLUT 1 glucose transporter elevation in the plasma membrane–enriched fraction, yet expressed a >2‐fold increase in glucose transport. Furthermore, the G14 cells had a similar content of GLUT 1 glucose transporter in the plasma membrane fraction when compared with the V79 parental cell line. Using Western blot analysis, the GLUT 1 glucose transporter in G14 cells exhibited a different mobility on a polyacrylamide gel when compared with the mobility of the GLUT 1 glucose transporter of the V79 cell line. This differential mobility of the glucose transporters in the hamster cells appeared to be related to glycosylation differences of the glucose transporters. Although normal human and hamster cell lines exhibited significant increases in insulin‐stimulated sugar transport (P < 0.05), the two respective respiration‐deficient cell lines exhibited no significant increases in insulin‐stimulated sugar transport (P > 0.05). Additionally, the expression of the GLUT 1 mRNA in the human WG750 mutant cells was elevated when compared with GLUT 1 mRNA in normal cells. Insulin exposure significantly increased GLUT 1 mRNA in human cells (P < 0.05). No differences in the GLUT 1 mRNA were observed between both hamster cell lines. Thus, both respiration‐deficient cell lines are insulin resistant (i.e., regarding their insulin‐stimulated sugar transport). The respiration‐deficient mutation results in an increased sugar transport in the human and hamster cells; however, the human cells adapt to the mutation by increasing their levels of GLUT 1 mRNA and eventually membrane‐located glucose transporters. On the other hand, the hamster cells adapt by apparently modifying their glucose transporters' intrinsic activity via glycosylation. We feel that these cell systems can be effective models to study the multiple factors involved in sugar transport regulation in vertebrate cells.

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Section: Resultsmentioning

confidence: 66%

mentioning

confidence: 59%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 97%

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Sugar Transport Regulation: Comparative Characterization of the Effect of NADH CoQ Reductase Deficiency in Two Cell Culture Systems

Germinario

Continelli

Pratt

2000

Proc Soc Exp Biol Med

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Organization of neural inputs to the suprachiasmatic nucleus in the rat

1997

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The circadian timing of the suprachiasmatic nucleus (SCN) is modulated by its neural inputs. In the present study, we examine the organization of the neural inputs to the rat SCN using both retrograde and anterograde tracing methods. After Fluoro-Gold injections into the SCN, retrogradely labeled neurons are present in a number of brain areas, including the infralimbic cortex, the lateral septum, the medial preoptic area, the subfornical organ, the paraventricular thalamus, the subparaventricular zone, the ventromedial hypothalamic nucleus, the posterior hypothalamic area, the intergeniculate leaflet, the olivary pretectal nucleus, the ventral subiculum, and the median raphe nuclei. In the anterograde tracing experiments, we observe three patterns of afferent termination within the SCN that correspond to the photic/raphe, limbic/hypothalamic, and thalamic inputs. The median raphe projection to the SCN terminates densely within the ventral subdivision and sparsely within the dorsal subdivision. Similarly, areas that receive photic input, such as the retina, the intergeniculate leaflet, and the pretectal area, densely innervate the ventral SCN but provide only minor innervation of the dorsal SCN. A complementary pattern of axonal labeling, with labeled fibers concentrated in the dorsal SCN, is observed after anterograde tracer injections into the hypothalamus and into limbic areas, such as the ventral subiculum and infralimbic cortex. A third, less common pattern of labeling, exemplified by the paraventricular thalamic afferents, consists of diffuse axonal labeling throughout the SCN. Our results show that the SCN afferent connections are topographically organized. These hodological differences may reflect a functional heterogeneity within the SCN.

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Circadian Rhythms and Depression: Clinical and Experimental Models

Rosenwasser¹,

Wirz‐Justice²

1997

Handbook of Experimental Pharmacology

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Permanent hexose transport upregulation in a respiration-deficient human fibroblast cell strain

Cited by 15 publications

References 28 publications

Sugar Transport Regulation: Comparative Characterization of the Effect of NADH CoQ Reductase Deficiency in Two Cell Culture Systems

Sugar Transport Regulation: Comparative Characterization of the Effect of NADH CoQ Reductase Deficiency in Two Cell Culture Systems

Organization of neural inputs to the suprachiasmatic nucleus in the rat

Circadian Rhythms and Depression: Clinical and Experimental Models

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