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Effects of sheep dip pesticides on differentiating nerve cells: Identification of novel markers of toxicity - VM02300

Description
The main objective of the work is to study key morphological and proteomic changes associated with exposure to sub-lethal concentrations of sheep dip pesticides. This will be achieved by the application of a combination of molecular and morphological approaches to the analysis of differentiating nerve cells and cell extracts prepared following exposure of cells to sheep dip pesticides both individually and in combination. Experiments will also be performed in the presence of activated rat liver microsomes or potential neuroprotective agents such as GDNF to study the reversibility of the observed sub-lethal effects.
Studies of MTT reduction will help to establish the concentration range over which cell viability is reduced. The application of sub-lethal concentrations will help to determine the effective concentration range causing inhibition of neurite outgrowth. Cell lysates and cytoskeletal extracts prepared under selected conditions of sub-lethal toxicity will be analysed by probing western blots with antibodies that recognise specific cytoskeletal and axon growth associated proteins. The effects of sub-lethal exposure on the organisation and cellular distribution of proteins of interest will be further analysed by immunostaining techniques.

Consistent changes in other proteins will be studied by 2D-PAGE and polypeptides of interest that show consistent changes following exposure will be excised from gels and analysed by mass spectrometry, after which sequence data will be compared to proteomic databases on the internet to establish identity.
Therefore, by using an in vitro alternative to animals, this study will therefore provide important information about the pattern of morphological and molecular changes that follow exposure of neuronal cells to sub-lethal levels of pesticides and how this is influenced by combined exposure, metabolic activation or neuroprotective agents. This, in turn, will help us to understand the mechanism(s) of long term sub-chronic effects of pesticides and pesticide mixtures, revealing important markers of toxicity that may also represent potential targets for the future development of therapeutic intervention strategies.
Project Documents
• Final Report : Effects of sheep dip pesticides on differentiating nerve cells: Identification of novel markers of toxicity   (677k)
Time-Scale and Cost
From: 2004

To: 2006

Cost: £158,751
Contractor / Funded Organisations
University - Nottingham Trent
Keywords
Animal Health              
Neuropathology              
Organophosphates              
Pesticides              
Plants and Animals              
Sheep Dipping              
Veterinary Medicines              
Fields of Study
Animal Health
Veterinary Medicine