Glucose transporter 3 (GLUT3) protein expression in human placenta across gestation.
Glucose transporter 3 (GLUT3) protein expression in human placenta across gestation.
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DOI：
10.1016/j.placenta.2011.09.014
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发表时间：
2011-12
期刊：
影响因子：
3.8
通讯作者：
Illsley, N. P.
中科院分区：
文献类型：
--
作者：
Brown, K.;Heller, D. S.;Zamudio, S.;Illsley, N. P.
关于人胎盘中GLUT 3蛋白表达的补充信息已有报道，但GLUT 3蛋白在整个妊娠期的表达定位和模式尚未明确定义。本研究的目的是表征整个妊娠期的合胞体GLUT 3蛋白表达。我们假设GLUT 3蛋白存在于合胞体微绒毛膜中，并且其表达在妊娠期间减少。在一系列胎龄（妊娠早期至妊娠晚期）的样品中测量GLUT 3蛋白，人脑和人肠分别用作阳性和阴性对照。作为特异性的额外测量，我们用针对GLUT 3的siRNA转染BeWo绒毛膜癌细胞（一种表达GLUT 3的滋养层细胞系），并通过Western印迹分析表达。免疫组化法检测到GLUT 3在所有胎龄的合体滋养细胞。使用蛋白质印迹法，GLUT 3被检测为一个完整的膜蛋白，分子量约为50 kDa的微绒毛膜从所有妊娠期，但不是在合胞基底膜。通过证明GLUT 3 siRNA处理的BeWo中免疫印迹信号的表达降低至用非靶向siRNA转染的细胞中观察到的表达的18 ± 6%（平均值± SEM）来确认一抗靶标的身份。通过蛋白质印迹检测的微绒毛膜中的GLUT 3表达在整个妊娠期降低，使得在妊娠中期（第14-26周）的表达为妊娠早期的表达的48 ± 7%，并且到妊娠晚期（第31-40周）仅为妊娠早期表达的34 ± 10%。此外，用GLUT 3 siRNA处理的BeWo细胞中的葡萄糖摄取减少到用非靶向siRNA处理的细胞中测量的葡萄糖摄取的60%。这表明GLUT 3介导的摄取占BeWo细胞葡萄糖摄取的约50%。这些结果证实了GLUT 3在妊娠早期存在于合胞体微绒毛膜中并在此后减少的假设，支持GLUT 3在妊娠早期对葡萄糖摄取更重要的想法。
Conflicting information regarding expression of GLUT3 protein in the human placenta has been reported and the localization and pattern of expression of GLUT3 protein across gestation has not been clearly defined. The objective of this study was characterization of syncytial GLUT3 protein expression across gestation. We hypothesized that GLUT3 protein is present in the syncytial microvillous membrane and that its expression decreases over gestation. GLUT3 protein was measured in samples from a range of gestational ages (first to third trimester), with human brain and human bowel used as a positive and negative control respectively. As an additional measure of specificity, we transfected BeWo choriocarcinoma cells, a trophoblast cell line expressing GLUT3, with siRNA directed against GLUT3 and analyzed expression by Western blotting. GLUT3 was detected in the syncytiotrophoblast at all gestational ages by immunohistochemistry. Using Western blotting GLUT3 was detected as an integral membrane protein at a molecular weight of ~50kDa in microvillous membranes from all trimesters but not in syncytial basal membranes. The identity of the primary antibody target was confirmed by demonstrating that expression of the immunoblotting signal in GLUT3 siRNA-treated BeWo was decreased to 18 ± 6% (mean ± SEM) of that seen in cells transfected with a non-targeting siRNA. GLUT3 expression in microvillous membranes detected by Western blot decreased through the trimesters such that expression in the second trimester (wks 14–26) was 48 ± 7% of that in the first trimester and by the third trimester (wks 31–40) only 34 ± 10% of first trimester expression. In addition, glucose uptake into BeWo cells treated with GLUT3 siRNA was reduced to 60% of that measured in cells treated with the non-targeting siRNA. This suggests that GLUT3-mediated uptake comprises approximately 50% of glucose uptake into BeWo cells. These results confirm the hypothesis that GLUT3 is present in the syncytial microvillous membrane early in gestation and decreases thereafter, supporting the idea that GLUT3 is of greater importance for glucose uptake early in gestation.
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3.8
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通讯作者：
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