RubyACRs enable red-shifted optogenetic inhibition in freely behaving Drosophila

Optogenetic neuronal activators with red-shifted excitation spectra, such as Chrimson, have significantly advanced Drosophila neuroscience. However, until recently, available optogenetic inhibitors required shorter activation wavelengths, which do not penetrate tissue as effectively and are stronger visual stimuli to the animal, potentially confounding behavioral results. Here, we assess the efficacy of two newly identified anion-conducting channelrhodopsins with spectral sensitivities similar to Chrimson: A1ACR and HfACR (RubyACRs). Electrophysiology and functional imaging confirmed that RubyACRs effectively hyperpolarize neurons, with stronger and faster effects than the widely used inhibitor GtACR1. Activation of RubyACRs led to circuit-specific behavioral changes in three different neuronal groups. In glutamatergic motor neurons, activating RubyACRs suppressed adult locomotor activity. In PPL1-γ1pedc dopaminergic neurons, pairing odors with RubyACR activation during learning produce..., , # RubyACRs enable red-shifted optogenetic inhibition in freely behaving Drosophila

Dataset DOI: [10.5061/dryad.gxd2547zg ](10.5061/dryad.gxd2547zg)respository

## Description of the data and file structure

This dataset describes RubyACR performance in Drosophila melanogaster with 5 different assays. These five assays correspond to the 5 figures in the accompanying paper. In each assay, there is a \"Data\" folder containing a pickle file(s) with the data and a \"Script\" folder containing Python Jupyter notebooks that generate the graphs found in the paper. The Python version and libraries used in the Python environment are described in the Pipfile and Pipfile.lock text files.

Figure 1 includes electrophysiological recordings from the larval NMJ motor neurons expressing RubyACR.

Figure 2 includes imaging from Mi1 neurons expressing RubyACR in the adult.

Figure 3 includes walking behavior when RubyACRs are activated in glutamatergic neurons.

Figure 4 includes memory responses when RubyA...,