OBJECTIVE: Our purpose was to determine the effects of insulin and glucose on glucose transport and expression of GLUT1 glucose transporter messenger ribonucleic acid in first-trimester human trophoblast-like cells. STUDY DESIGN: First-trimester human trophoblast-like cells were maintained as a continuous cell line. For 2[3H]deoxy-d-glucose uptake and messenger ribonucleic acid studies the cells were incubated in the presence or absence of insulin (10-7 to 10-11 mol/L) or d-glucose (0 to 50 mmol/L) for 0 to 24 hours. Glucose transport was measured by incubating cells with 0.1 mmol/L,2[3H]deoxy-d-glucose for 5 minutes. Specific uptake was determined by incubating companion cultures with 10 μmol/L cytochalasin B. The cells were then solubilized with sodium hydroxide and the radioactivity counted. Data were expressed as nanomoles of 2[3H]deoxy-d-glucose transported per milligram of protein per 5 minutes and analyzed by one-way analysis of variance with post hoc testing by the method of Tukey. GLUT1 messenger ribonucleic acid was measured by Northern blotting of total ribonucleic acid samples hybridized to a phosphorus 32-labeled complementary deoxyribonucleic encoding the rat GLUT1 glucose transporter. As a control for loading efficiency, blots were stripped and rehybridized to a 40-mer phosphorus 32-labeled β-actin oligonucleotide probe. RESULTS: Insulin treatment resulted in a dose-dependent increase in the transport of 2[3H]deoxy-d-glucose at 24 hours (p < 0.001 at 10-7 mol/L). This change was first detected at 12 hours of incubation. These data closely paralled the insulin-induced increase in GLUT1 messenger ribonucleic acid seen in Northern blots. In contrast to insulin, increasing concentrations of d-glucose did not change the transport of 2[3H]deoxy-d-glucose. However, when cells were incubated in low concentrations of d-glucose (0 or 1 mmol/L), an enhancement in the uptake of 2[3H]deoxy-d-glucose (p < 0.001) was observed. Kinetic studies indicated that d-glucose augmentation of 2[3H]deoxy-d-glucose uptake was significant at 9 hours (p < 0.05). The effects of d-glucose on GLUT1 messenger ribonucleic acid expression paralleled the uptake of 2[3H]deoxy-d-glucose, although the modulation of GLUT1 messenger ribonucleic acid levels by glucose was much less pronounced than in insulin-treated cells. CONCLUSION: Although it has been assumed that the placenta has a limited role in influencing glucose transport to the fetus, our in vitro data demonstrate that both insulin and glucose can modulate glucose transport at the cellular level of the placental trophoblast. Thus maternal insulin and glycemic status may influence the expression of GLUT1, the major trophoblast glucose transporter protein, therefore directly affecting first-trimester placental glucose transport. These in vitro data may help explain the association between maternal glucose abnormalities and impaired fetal development during the first trimester when placental GLUT1 messenger ribonucleic acid expression is at its peak.
| Identifer | oai:union.ndltd.org:ETSU/oai:dc.etsu.edu:etsu-works-12596 |
| Date | 01 January 1995 |
| Creators | Gordon, Michael C., Zimmerman, Peter D., Landon, Mark B., Gabbe, Steven G., Kniss, Douglas A. |
| Publisher | Digital Commons @ East Tennessee State University |
| Source Sets | East Tennessee State University |
| Detected Language | English |
| Type | text |
| Source | ETSU Faculty Works |
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