Transgenic line for the identification of cholinergic release sites in Drosophila melanogaster.

Alexander Borst,Katarina Pankova

doi:10.1242/jeb.149369

Alexander Borst, Katarina Pankova

Open Access

https://doi.org/10.1242/jeb.149369

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Abstract

ABSTRACTThe identification of neurotransmitter type used by a neuron is important for the functional dissection of neuronal circuits. In the model organism Drosophila melanogaster, several methods for discerning the neurotransmitter systems are available. Here, we expanded the toolbox for the identification of cholinergic neurons by generating a new line FRT-STOP-FRT-VAChT::HA that is a conditional tagged knock-in of the vesicular acetylcholine transporter (VAChT) gene in its endogenous locus. Importantly, in comparison to already available tools for the detection of cholinergic neurons, the FRT-STOP-FRT-VAChT::HA allele also allows for identification of the subcellular localization of the cholinergic presynaptic release sites in a cell-specific manner. We used the newly generated FRT-STOP-FRT-VAChT::HA line to characterize the Mi1 and Tm3 neurons in the fly visual system and found that VAChT is present in the axons of both cell types, suggesting that Mi1 and Tm3 neurons provide cholinergic input to the elementary motion detectors, the T4 neurons.

Highlights

Understanding of the information processing in neuronal circuits requires knowledge about connectivity and properties of the cells involved
The position of the tag was chosen such that it would not interfere with protein folding or signaling sequences known to participate in the intracellular trafficking of vesicular acetylcholine transporter (VAChT) (Fei et al, 2008)
Our aim was to restrict the expression of VAChT::HA to a population of neurons defined by the expression pattern of a Gal4 line of choice

Summary

Introduction

Understanding of the information processing in neuronal circuits requires knowledge about connectivity and properties of the cells involved. The type of neurotransmitter released by a cell defines, to a large extent, the role of a cell and the range of logical operations that are performed within a circuit. The identification of the cellular neurotransmitter phenotype is of crucial importance for the functional dissection of neuronal circuits. Various techniques have been described for the identification of neurotransmitter systems in the Drosophila melanogaster nervous system. In Drosophila neurons, the major disadvantage of this strategy is that

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