Published June 2, 2022 | Version v1

Population and single dopamine neuron activity during classical conditioning

  • 1. Harvard University

Description

We trained naive or trained mice to associate odor cues with outcome (water, air puff or no outcome), and recorded dopamine cell body activity in the vetral tegmental area (VTA), dopamine axon activity in the ventral striatum (VS) or dopamine release in VS with optic fiber fluorometry (photometry). In different set of mice, single dopamine neuron activiy was recorded with 2-photon microscope. In some of these mice, we reversed odor-outcome contingency so that an odor that was associated with no outcome or air puff became associated with water reward. Licking pattern was also recorded.

Notes

Fiber-fluorometry 

The noise from the power line in the voltage signal was cleaned by removing 58-62Hz signals through a band stop filter. Z-score was calculated from signals in an entire session smoothed with moving average of 50 ms. To average data using different sessions, signals were normalized as follows. The global change of signals within a session was corrected by linear fitting of signals and time and subtracting the fitted line from signals. The baseline activity for each trial (F0each) was calculated by averaging activity between -1 to 0 sec before a trial start (odor onset for odor trials and water onset for free water trials), and the average baseline activity for a session was calculated by averaging F0each of all trials (F0average). dF/F was calculated as (F - F0each)/F0average. dF/F was then normalized by dividing by average responses to free water (0 to 1 sec from water onset). 

2-photon imaging

Acquired images were preprocessed in the following manner: 1) movement correction was performed using phase correlation image registration implemented in Suite2P59; 2) region-of-interest (ROI) selection was performed manually from the mean and standard deviation projections of a subset of frames from the entire acquisition, as well as a movie of the frames used to build those projections; 3) Neuropil decontamination was performed with FISSA60 using four regions around each ROI. The decontaminated signal extracted from these regions was then flattened (to correct for drifts caused by photobleaching), and z-scored across the entire session. The preprocessed and z-scored fluorescence were resampled to 30 Hz and used for further analysis. The first one was used to calculate ∆F/F0 in which F0 was the mean fluorescence before odor delivery. When the z-scored fluorescence was used for analysis, the mean z-score value before odor delivery was subtracted.

Funding provided by: National Institute of Mental Health
Crossref Funder Registry ID: http://dx.doi.org/10.13039/100000025
Award Number: R01MH125162

Funding provided by: National Institute of Neurological Disorders and Stroke
Crossref Funder Registry ID: http://dx.doi.org/10.13039/100000065
Award Number: U19 NS113201

Funding provided by: National Institute of Neurological Disorders and Stroke
Crossref Funder Registry ID: http://dx.doi.org/10.13039/100000065
Award Number: NS 108740

Funding provided by: Simons Foundation
Crossref Funder Registry ID: http://dx.doi.org/10.13039/100000893
Award Number: the Simons Collaboration on Global Brain

Funding provided by: Japan Society for the Promotion of Science
Crossref Funder Registry ID: http://dx.doi.org/10.13039/501100001691
Award Number: Japan Science and Technology Agency

Funding provided by: Human Frontier Science Program
Crossref Funder Registry ID: http://dx.doi.org/10.13039/501100000854
Award Number: LT000801/2018

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