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Purification and microRNA Profiling of Exosomes Derived from Blood and Culture Media
Published on: June 14, 2013
Selenoprotein mRNA is expressed in blood at levels comparable to major tissues in rats
Jacqueline K Evenson1, Adam D Wheeler, Sean M Blake
1Department of Nutritional Sciences, University of Wisconsin, Madison, WI 53706, USA.
Abstract:
Liver glutathione peroxidase-1 (GPX1) mRNA is highly regulated by Se status relative to other parameters, but is of limited use for determining Se requirements in humans. To examine the efficacy of using blood for Se status assessment using molecular biology markers, we used a ribonuclease protection assay (RPA) to study mRNA levels in whole blood relative to 16 other rat tissues. Significant amounts of total RNA (>50 microg) were obtained from 1 mL of whole blood. Total RNA from 28-d postweaning Se-adequate (0.2 microg Se/g diet) male rats was analyzed for GPX1, GPX4, GPX3, thioredoxin reductase-1 (TRR1), and selenoprotein-P (SelP). RPA detected significant mRNA expression for at least 1 selenoprotein in all tissues except pancreas. GPX1 mRNA expression using this mix of RPA probes yielded the highest signal for GPX1 relative to the other selenoprotein signals in all tissues except testis; GPX1 expression was 4th highest in blood and similar to the major organs (liver, 1st; heart, 5th; kidney, 6th). Kidney was highest for GPX3, and testes was highest for GPX4, TRR1, and SelP. This study is the first to report the gene expression pattern for a number of selenoproteins and across a comprehensive set of tissues. The mRNA levels for all selenoproteins in blood were comparable to levels in the major organs, and decreases in blood and liver GPX1 mRNA levels in Se deficiency were similar, supporting potential use of whole blood for assessing Se status using molecular biology markers.
Insights
Whole blood can effectively assess selenium (Se) status using molecular markers. Glutathione peroxidase-1 (GPX1) mRNA levels in blood mirror liver levels, indicating its utility for Se status evaluation.
Area of Science:
- Biochemistry
- Molecular Biology
- Nutritional Science
Background:
- Liver glutathione peroxidase-1 (GPX1) mRNA is sensitive to selenium (Se) status but not ideal for human requirements.
- Assessing Se status via molecular markers in blood is a potential alternative.
Purpose of the Study:
- To evaluate the efficacy of using whole blood for Se status assessment via molecular biology markers.
- To analyze selenoprotein mRNA expression patterns across various rat tissues.
Main Methods:
- Ribonuclease protection assay (RPA) was used to quantify mRNA levels of GPX1, GPX4, GPX3, thioredoxin reductase-1 (TRR1), and selenoprotein-P (SelP) in whole blood and 16 other rat tissues.
- Total RNA was extracted from 1 mL of whole blood from Se-adequate male rats.
Main Results:
- Significant selenoprotein mRNA expression was detected in all tissues except the pancreas.
- GPX1 mRNA was highly expressed, with levels in blood comparable to major organs like the liver.
- Decreases in GPX1 mRNA in blood and liver were similar under Se deficiency.
Conclusions:
- Whole blood is a viable source for assessing Se status using molecular markers.
- The gene expression patterns of selenoproteins across tissues provide a comprehensive reference.
- Blood GPX1 mRNA levels serve as a reliable indicator of Se status, similar to liver levels.
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