Analytical and Bioanalytical Chemistry

, Volume 397, Issue 8, pp 3369–3375

Evaluating the effects of estradiol on endothelial nitric oxide stimulated by erythrocyte-derived ATP using a microfluidic approach

  • Suzanne Letourneau
  • Luiza Hernandez
  • Andrea N. Faris
  • Dana M. Spence
Original Paper

DOI: 10.1007/s00216-010-3687-7

Cite this article as:
Letourneau, S., Hernandez, L., Faris, A.N. et al. Anal Bioanal Chem (2010) 397: 3369. doi:10.1007/s00216-010-3687-7

Abstract

Recently, estrogens have been reported to have protective effects against experimental autoimmune encephalomyelitis (EAE), a murine model of multiple sclerosis (MS). Although the molecular mechanism for such a protective effect is currently incomplete, we hypothesized that estradiol may reduce the release of ATP from erythrocytes (ERYs), thereby lowering the production of nitric oxide (NO) by endothelial cells. Here, we report on the use of a microfluidic device to investigate the direct effects of the estrogen estradiol on endothelial cell nitric oxide production. In addition, the incorporation of a thin polycarbonate membrane into the device enabled the passage of ERYs through the device to determine indirect effects of estradiol on NO production that may be meditated by ERYs. When these ERYs were incubated with increasing concentrations of estradiol, the NO production from the endothelial cells was attenuated to a value that was only 59 ± 7% of ERYs in the absence of estradiol. This decrease in NO production coincides with reductions in ERY-derived ATP release in the presence of estradiol. Estradiol is typically reported to have NO-stimulating effects; however, such reports have employed in vitro experimental designs that include only a single cell type. To demonstrate the potential importance of this attenuation of ATP from ERYs, results from a small-scale study show that the ATP release obtained from healthy controls was 138 ± 21 nM (n = 18) while the release from the ERYs obtained from people with MS was 375 ± 51 nM (n = 11). The studies reported here involving multiple cells types (endothelial cells and ERYs) may lead to a reappraisal of the in vivo activities of estradiol.

Keywords

Bioanalytical methodsBiological samplesBiotechnological productsCell systems/single-cell analysisMicrofluidics/microfabricationFluorescence/luminescence

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Suzanne Letourneau
    • 1
  • Luiza Hernandez
    • 1
  • Andrea N. Faris
    • 1
  • Dana M. Spence
    • 1
  1. 1.Department of ChemistryMichigan State UniversityEast LansingUSA