Carbon fiber microelectrode amperometry (CFMA) is explored as a technique for studying the effects of immunotoxicants on single-cell in vitro exocytosis function in a mouse peritoneal mast cell (MPMC)/fibroblast co-culture model. MPMCs were acutely exposed to between 10 and 100 μM of the immunotoxicants mono-2-ethylhexyl phthalate (MEHP) and bisphenol A (BPA), and release of serotonin was evaluated by CFMA. A significant decrease in the quantal content of serotonin was measured for all levels of exposure to both MEHP and BPA. The overall efficiency of the exocytotic function of MPMCs was found to be impaired by all exposure concentrations of BPA, but this efficiency was only impaired at the lowest exposure concentration of MEHP. This study illustrates the potential of CFMA as a technique for determining quantitative and biophysical chemical information in in vitro immunotoxicological studies. [Figure not available: see fulltext.]
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variation in Q from granule-to-granule within that cell. BPA exposure (left) shows a decrease in Q with increased exposure time while MEHP exposure (right) and controls show no exposure time dependence in quantal release Acknowledgements Financial support for this work was generously provided by the National Science Foundation (no. CHE-0645041). All electron microscopy work was carried out in the Institute of Technology Characterization Facility, University of Minnesota, which receives partial support from NSF through the NNIN program. We would also like to acknowledge Alice Ressler of the Technology Characterization Facility and former student Eva Cornell for their assistance in developing and performing the TEM sample preparation.
Copyright 2011 Elsevier B.V., All rights reserved.
- Bioanalytical methods
- Cell systems
- Electroanalytical methods
- Endocrine disruptors
- Exocytosis microelecrtodes
- Single-cell analysis