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ToxSci Advance Access published online on October 5, 2005
Toxicological Sciences, doi:10.1093/toxsci/kfj003
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© The Author 2005. Published by Oxford University Press on behalf of the Society of Toxicology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org
Received August 24, 2005
Accepted September 22, 2005

Reproductive and Developmental Toxicology

Exposure to Diazinon Alters In Vitro Retinogenesis: Retinospheroid Morphology, Development of Chicken Retinal Cell Types and Gene Expression

L. E. Paraoanu 1*, J. B. Mocko 1, M. Becker-Roeck 1, J. Smidek-Huhn 1, and P. G. Layer 1

1 University of Technology Darmstadt, Institute of Zoology, Department of Developmental Biology and Neurogenetics Schnittspahnstrasse 3, D-64287, Darmstadt, Germany

* To whom correspondence should be addressed.
L. E. Paraoanu, E-mail: paraoanu{at}bio.tu-darmstadt.de


  Abstract

Developing embryos are more vulnerable than adults to acute cholinergic intoxication by anticholinesterases, including organophosphorus pesticides. These agents affect the process of neural development itself, leading to permanent deficits in the architecture of the nervous system. Recent evidence on direct roles of acetylcholinesterase (AChE) on neuronal differentiation provides additional grounds for investigating the developmental toxicity of anticholinesterases. Therefore, the effect of the organophosphate diazinon on the development of chick retinal differentiation was studied by an in vitro reaggregate approach. Reaggregated spheres from dissociated retinal cells of the E6 chick embryo were produced in rotation culture. During the whole culture period of 10 days, experimental cultures were supplemented with different concentrations of the pesticide, from 20 to 120 µM diazinon. The pesticide-treated spheres were reduced in size, and their outer surface was irregular. More importantly, inner structural distortions could be easily traced, since the structure of control spheroids can be well characterised by a histotypical arrangement of laminar parts homologous to the normal retina. AChE activity in diazinontreated spheres was reduced when compared with controls. As a dramatic effect of exposure to the pesticide, IPL-like areas in spheroids were not distinguishable anymore. Similarly, photoreceptor rosettes and Müller radial glia were strongly decreased, while apoptosis was stimulated. The expression of transcripts for choline-acetyltransferase and muscarinic receptors was affected, revealing an effect of diazinon on the cholinergic system. This further proves the significance of cholinesterases and the cholinergic system for proper nervous system development and shows that further studies of debilitating diazinon actions on development are necessary.


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