Premium
Effects of di‐n‐butyl phthalate on neuron development (1139.1)
Author(s) -
Feng Josh,
Acey Roger
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.1139.1
Subject(s) - phthalate , chemistry , hsp70 , gel electrophoresis , heat shock protein , cell , microbiology and biotechnology , biochemistry , biology , gene , organic chemistry
Plasticizers such as di‐n‐butyl phthalate (DBP) and bisphenol A (BPA) are well‐documented, ubiquitously distributed environmental contaminants. The effect of these compounds on human health has been controversial but research has strongly suggested they function as estrogen mimics or “endocrine disruptors”. We have developed a human umbilical cord mesenchymal stem cell (HUC‐SC) system as a model to predict relative toxicities of phthalate esters. The default pathway of these differentiating cells is the formation of neural tissue. Differentiating cells are exposed to the potential toxin for increasing periods of time, collected, and homogenized. Cell lysates are then analyzed by two‐dimensional polyacrylamide gel electrophoresis (2D‐PAGE) for changes in protein expression. Our first series of experiments looked at the effect of 10 nM DBP on undifferentiated HUC‐SCs. At this concentration of phthalate, there was no effect on cell proliferation, membrane integrity, or induction of apoptosis. Next, the HUC‐SCs were induced to differentiate in the presence or absence of 10 nM DBP. Cells were collected after 20, 24, 28, 32, and 48 hours of culture and analyzed by 2D‐PAGE. Comparison of control against phthalate‐treated cell extracts showed dramatic differences in protein expression. Preliminary analysis by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectroscopy (MALDI‐TOF MS) reveals two significant differentially‐expressed proteins: enolase, which suggests that DBP may be altering the neuron differentiation pathway, and heat shock protein β1, indicating that the cells are under stress. Future experiments will be to identify other differentially expressed proteins and determine their potential as biomarkers for evaluating the extent of DBP exposure. Grant Funding Source : Supported by the Doris A. Howell‐CSUPERB Research Scholars Award Program