Unique functional responses differentially map onto genetic subtypes of dopamine neurons

Dopamine neurons are characterized by their response to unexpected rewards, but they also fire during movement and aversive stimuli. Dopamine neuron diversity has been observed based on molecular expression profiles; however, whether different functions map onto such genetic subtypes remains unclear. In this study, we established that three genetic dopamine neuron subtypes within the substantia nigra pars compacta, characterized by the expression of Slc17a6 (Vglut2), Calb1 and Anxa1, each have a unique set of responses to rewards, aversive stimuli and accelerations and decelerations, and these signaling patterns are highly correlated between somas and axons within subtypes. Remarkably, reward responses were almost entirely absent in the Anxa1+ subtype, which instead displayed acceleration-correlated signaling. Our findings establish a connection between functional and genetic dopamine neuron subtypes and demonstrate that molecular expression patterns can serve as a common framework to dissect dopaminergic functions.


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All manuscripts must include a data availability statement.This statement should provide the following information, where applicable: -Accession codes, unique identifiers, or web links for publicly available datasets -A description of any restrictions on data availability -For clinical datasets or third party data, please ensure that the statement adheres to our policy A whole section of the methods is dedicated to explaining excluding criteria in detail.In brief, fiber photometry recordings were excluded if transient signal-to-noise ratios were below a threshold, which occurred when recordings were made in regions without sufficient axons or somas of the subtype of interest.This threshold was determined independently of the subsequent analysis steps.Recordings were also excluded if movement artifacts were detected in the 405 nm isosbestic control.For behavioral analysis, recordings where mice were not running for at least 100s were excluded from locomotion analysis, and rewards delivery times were the mice did not lick immediately after the reward was delivered were also excluded from analysis.
The new Anxa1+ subtype was identified from analysis of both a meta-dataset of existing single-cell RNAseq and a new dataset of single-nucleus RNAseq.Subtype marker expression was corroborated using the Allen Brain Atlas in situ hybridization dataset.Locomotion signaling was corroborated using several complimentary analysis (cross-correlation with acceleration and different triggered averages.PCA analysis and reward/air puff signaling were corroborated using different normalization settings.Functional analysis of different subtypes was corroborated in recordings from striatum and SNc.
The same experiments and measures were made from different dopaminergic subtypes such that other subtypes served as controls to each The same experiments and measures were made from different dopaminergic subtypes such that other subtypes served as controls to each other.We also repeated the experiments in DAT-Cre mice where all subtypes were simultaneously recorded from, as an additional control.other.We also repeated the experiments in DAT-Cre mice where all subtypes were simultaneously recorded from, as an additional control.Thus, group randomization was not necessary.Thus, group randomization was not necessary.
Data collection and analysis were not performed blind to the conditions of the experiments, as different subtypes require recording from different (though Data collection and analysis were not performed blind to the conditions of the experiments, as different subtypes require recording from different (though partially overlapping) regions of striatum, due to their different projection regions.Exclusion criteria were selected blind subtype identity and subsequent analysis.partially overlapping) regions of striatum, due to their different projection regions.Exclusion criteria were selected blind subtype identity and subsequent analysis.

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