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Fish Indicatiors of Stress and Health (FISH): Validating the use of embryos and yolk sac larval zebrafish as a model for the study of nociception and veterinary drug testing (FISH)
Start date: Apr 14, 2014, End date: Apr 13, 2016 PROJECT  FINISHED 

The zebrafish (ZF) is now a widely accepted vertebrate model for research in genetics, neuroscience and development due to its sheer experimental convenience and also its gene, developmental and functional homologies with mammals. ZF are also exploited as a novel model in nociceptive or pain research in a number of laboratories. We are already assessing behavioural and physiological responses in adult ZF to pain but wish to also explore whether embryonic and larval forms offer a useful alternative to adults. This project has three stages. First, using a sophisticated video capture system and automated computational analysis of behaviour, we will compare the behavioural responses to different pain modalities of non-protected larval stages with adults from our current NC3Rs study. This will establish quantifiable behavioural responses to potentially painful stimulation in the larval model. Second, we will characterise the molecular pathways of nociception through a genome-wide analysis of gene expression of the central nervous system (CNS). The screen will be implemented using RNASeq on Illumina and results will be correlated with behaviour. The functional impication of gene responses will be explored by modelling responding genes within a zebrafish regulatory gene network model which will be compared with equivalent mammalian data. We will validate a selection of known and novel candidates by PCR and further in situ hybridisation will determine the precise location of expression in the CNS and adult brain producing a 3D map of brain activity using light sheet microscopy. A third objective is to test a range of analgesic drugs to define which are the most effective in reducing the pain-related behavioural responses and to establish a reliable protocol for wider analgesic testing. The resulting data will be made publically available to researchers, biomedical scientists and veterinarians in order to improve welfare and reduce numbers of adult fish used.

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