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Ontogenesis of visual processing circuits in the zebrafish: analysis of specific neuronal subpopulations in development
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The Zebrafish (Danio rerio) larva is an ideal model for identification of neuronal circuits such as those
associated with visually guided behaviors like the optomotor and the optokinetic responses. The use of
larvae expressing the calcium level indicator GCaMP in particular neuronal subpopulations allows
monitoring of activity of these neurons and facilitates their anatomical and functional characterization.
Our goal, in this context, is to characterize some of these subpopulations taking advantage of the
expression of GFP using the Gal4-UAS system during the first 6 days of development. To examine and
characterize the GFP expression in zebrafish larvae, we have applied an immunofluorescence protocol
and recorded anatomical stacks using confocal imaging. This information gathered from several
equivalent individuals and registered to a reference brain may then be used to establish anatomical
atlases at these early stages of development that will complement those already developed for the 6 dpf
larvae. Characterization of the transgenic lines at the early stages will help us to locate the functional
clusters identified at 6 dpf with more anatomical accuracy and precision. We will present our work
focusing on zebrafish lines, generated using a BAC transgenic approach, that expresses GFF or GFP
from an insertion next to the olig2, chrna4 and pcp4a promoter regions. Two of these lines (olig2: GFP
and chrna4: GFF) are shown to represent a similar pattern of expression has the original gene, whilst the
pcp4a: GFF line does not, although its characterization is still of use to complement functional maps of
neuron activity. In addition, we are using time lapse imaging with Lightsheet microscopy to follow the
dynamics of neuronal differentiation and extension of projections during the 20-48 hours post
fertilization period on an already characterized line(Chat:GFF).
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Zebrafish olig2 chrna4 pcp4a pitx2c chat Confocal