Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2009 Jan;10(1):37-49.
doi: 10.3390/ijms10010037. Epub 2008 Dec 27.

Developmental toxicity of Ochratoxin A in rat embryo midbrain micromass cultures

Iwona Wilk-Zasadna  1 , Maria Minta  1
Affiliations

Developmental toxicity of Ochratoxin A in rat embryo midbrain micromass cultures

Iwona Wilk-Zasadna et al. Int J Mol Sci. 2009 Jan.

Abstract

Embryonic midbrain micromass cultures were exposed for five days to ochratoxin A (OTA) at seven concentrations (ranging from 0.16 to 10 microg/mL). Cell viability was assessed in neutral red uptake test (NRU), and differentiation - by immunoenzymatic determination of structural proteins (beta(III)-tubulin, MAP2, GFAP) expression level as well as by computer image analysis. Dose dependent decrease in cell number and differentiation was observed. Concentration-response curves were analysed and the mean inhibition concentrations (microg/mL) for cytotoxicity (IC(50)) and differentiation (ID(50)) were calculated. There were no significant differences in the sensitivity of neurons in early and late stage of differentiation and astrocytes to the toxic activity of this compound. For all endpoints ID(50) value was very low (< 10 microg/mL) so OTA was classified as a strong teratogen. IC(50)/ ID(50) ratios <2 pointed out that with harmful action of OTA the basic cytotoxicity should be connected.

Keywords: Ochratoxin A; computer image analysis; developmental neurotoxicity; embryonic midbrain cells; immunocytochemistry; in vitro micromass cultures.

PubMed Disclaimer

Figures

Figure 1.
Figure 1.
Photomicrographs showing control neurospheres stained using FITC- conjugated antibody against βIII tubulin, MAP2 and GFAP.
Figure 2.
Figure 2.
Representative concentration-response curves illustrating an effect of ochratoxin A on cytotoxicity (NRU test) and differentiation (six different endpoints) in micromass embryo midbrain cells cultured in vitro. The cytotoxic concentration of 2.5 μg/mL, mentioned in Table 1, resulted in increasing the number of neurospheres and decreasing of their area (B).
Figure 3.
Figure 3.
Images of midbrain micromass cultures, exposed for 5 days to different concentrations of OTA. Neurospheres were stained with haematoxylin (16 × magnification). After exposure to 2.5 μg/mL neurospheres were smaller and more numerous, after two highest concentrations no differentiated cells were observed.
See this image and copyright information in PMC

References

    1. Rodier PM. Developing brain as a target of toxicity. Environ. Health Perspect. 1995;103(suppl 6):73–76. - PMC - PubMed
    1. Grandjean P, Lanndrigan PJ. Developmental neurotoxicity of industrial chemicals. Lancet. 2006;368:2167–2178. - PubMed
    1. Mendola P, Selevan SG, Gutter S, Rice D. Environmental factors associated with spectrum of neurodevelopmental deficits. Ment. Retard. Dev. Disabil. Res. Rev. 2002;8:188–197. - PubMed
    1. Ceccatelli S, Tamm R, Tofighi R, Johansson C. In vitro systems to study developmental neurotoxicity of food contaminants. Toxicol. Lett. 2006;164(suppl):24. - PubMed
    1. Stoev SD. The role of ochratoxin A as a possible cause of Balkan endemic nephropathy and its risk evaluation. Vet. Hum. Toxicol. 1998;40:352–360. - PubMed

LinkOut - more resources