Description of the data and file structure Data Description
This dataset was collected from four cohorts of voles:
Sexually naive prairie voles (NP)
Sexually naive meadow voles (NM)
Pairbonded prairie voles (PP)
Paired meadow voles (PM)
Data was obtained through microscopy image analysis using Fiji ImageJ software. Data Collection & Processing Microscopy Analysis
DAPI masks were generated using the Analyze Particles function in Fiji ImageJ. Each region of interest (ROI) represents a nucleus.
The presence or absence of transcript was assessed for each ROI across three channels: D1DR (Drd1), D2DR (Drd2), and OXTR (Oxtr).
Data collected per ROI included:
ROI number
ROI area
Mean, minimum, and maximum gray values
Percent area covered by pixels
Oxtr Puncta Quantification
Oxtr puncta counts were recorded in the Oxtr Puncta Counts file for all four cohorts.
Secondary Object Count (Sec_Object Count) was collected only for the OXTR channel, representing the number of puncta per ROI.
Puncta detection utilized:
Morphological Filters Plugin (GitHub link)
Object Inspector (2D/3D) Plugin
Regional Classification
Each ROI was assigned a Region1 category:
Core1 = Core of the nucleus accumbens
Shell1 = Medial shell
Shell3 = Lateral shell
The Region variable specifies subregion and hemisphere (left/right).
Rostral-caudal levels (RC levels) of the nucleus accumbens were recorded.
Methods (From Manuscript)
Images were analyzed using Fiji ImageJ software (version 2.14.0/1.54f). Images were split into four channels:
Channel 0 = DAPI
Channel 1 = Drd1
Channel 2 = Drd2
Channel 3 = Oxtr
Regions of interest (ROIs) outlining DAPI-stained nuclei were automatically generated in Fiji ImageJ. Thresholds for DAPI nuclear staining were manually established to eliminate background and accurately overlay nuclei. DAPI Mask Validation
Accuracy was verified in 10% of images (one per animal) by comparing experimenter-counted vs. automatic nuclei counts.
Counts differed by ≤5%, supporting the reliability of automatic ROI generation.
Signal Detection & Quantification
The DAPI mask overlay was applied to Drd1, Drd2, and Oxtr images (Fig. 1C).
A white top-hat transformation enhanced contrast, improving bright feature detection.
Collected signal data included:
ROI number (nucleus number)
Minimum, mean, and maximum intensity values
% area (for both 16-bit and 8-bit data)
Data Processing & Analysis
Cellular distribution data was analyzed via custom Python code (GitHub repository).
This script identified:
Positively labeled nuclei for each channel
Double and triple-labeled cells (co-expression)
Oxtr Puncta Quantification
Oxtr puncta were quantified using:
Morphological Filters Plugin (GitHub link)
Object Inspector (2D/3D) Plugin
Files and variables File: R3_L1_PM_cellular_distribution_8_bit_v2.xlsx
Description: rostral-caudal level 1, paired meadow voles, transcript distribution data (8 bit) Variables
Region1 (nucleus accumbens subregion),
Region (nucleus accumbens subregion with laterality indicated),
Channel (label for transcript examined in that wavelength) ,
ROI (region of interest = nucleus) ,
Area (of nucleus/ROI),
Mean (gray value of transcript in that ROI in that channel indicated),
Min (gray value of transcript in that ROI in that channel indicated),
Max (gray value of transcript in that ROI in that channel indicated),
% Area (amount of area with nonzero pixels within ROI for that channel),
Sec Object Count (Oxtr puncta for OXTR channel only). Values are blank for D1DR and D2DR channels in this column.
File: R3_L1_NM_cellular_distribution_8_bit_v2.xlsx
Description: rostral-caudal level 1, naive meadow voles, transcript distribution data (8 bit) Variables
Region1 (nucleus accumbens subregion),
Region (nucleus accumbens subregion with laterality indicated),
Channel (label for transcript examined in that wavelength) ,
ROI (region of interest = nucleus) ,
Area (of nucleus/ROI),
Mean (gray value of transcript in that ROI in that channel indicated),
Min (gray value of transcript in that ROI in that channel indicated),
Max (gray value of transcript in that ROI in that channel indicated),
% Area (amount of area with nonzero pixels within ROI for that channel),
Sec Object Count (Oxtr puncta for OXTR channel only). Values are blank for D1DR and D2DR channels in this column.
File: R3_L1_NP_cellular_distribution_8_bit_v2.xlsx
Description: rostral-caudal level 1, naive prairie voles, transcript distribution data (8 bit) Variables
Region1 (nucleus accumbens subregion),
Region (nucleus accumbens subregion with laterality indicated),
Channel (label for transcript examined in that wavelength) ,
ROI (region of interest = nucleus) ,
Area (of nucleus/ROI),
Mean (gray value of transcript in that ROI in that channel indicated),
Min (gray value of transcript in that ROI in that channel indicated),
Max (gray value of transcript in that ROI in that channel indicated),
% Area (amount of area with nonzero pixels within ROI for that channel),
Sec Object Count (Oxtr puncta for OXTR channel only). Values are blank for D1DR and D2DR channels in this column.
File: R3_L1_PP_cellular_distribution_8_bit_v2.xlsx
Description: rostral-caudal level 1, pairbonded prairie voles, transcript distribution data (8 bit) Variables
Region1 (nucleus accumbens subregion),
Region (nucleus accumbens subregion with laterality indicated),
Channel (label for transcript examined in that wavelength) ,
ROI (region of interest = nucleus) ,
Area (of nucleus/ROI),
Mean (gray value of transcript in that ROI in that channel indicated),
Min (gray value of transcript in that ROI in that channel indicated),
Max (gray value of transcript in that ROI in that channel indicated),
% Area (amount of area with nonzero pixels within ROI for that channel),
Sec Object Count (Oxtr puncta for OXTR channel only). Values are blank for D1DR and D2DR channels in this column.
File: R3_L1_Oxtr_Puncta_Counts_v2.xlsx
Description: Oxtr puncta counts per ROI for naive prairie voles, naive meadow voles, pairbonded prairie voles, and paired meadow voles Variables
Animal # (sex is M or F and number is corresponding number given in colony)
Species (meadow or prairie vole)
Bond status (sexually naive or paired(bonded)
RC level (rostral caudal of nucleus accumbens; only level 1 is in this data set)
Region1 (subregion of nucleus accumbens)
Region (subregion of nucleus accumbens with laterality indicated)
ROI (region of interest corresponds to each nuclei detected)
Sec Object Count (# of Oxtr puncta detected within ROI)
Volume (^3) (data not used)
Code/software
Data files are in Excel sheets. Data sorting was done through Python scripts found on Donaldson Lab Github Citation 0 views 0 downloads 0 citations License: Public domain Subject keywords
Biological sciences dopamine receptor meadow vole oxytocin receptor pair bonding prairie vole
Funding
National Institutes of Health
U.S. National Science Foundation
Dana Foundation
Whitehall Foundation
Related works Preprint
https://doi.org/10.1101/2025.03.03.640889
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