Additional file 1 of Chromosomal and gonadal sex have differing effects on social motivation in mice
Creators: name: Chaturvedi, Sneha M.
name: Sarafinovska, Simona
name: Selmanovic, Din
name: McCullough, Katherine B.
name: Swift, Raylynn G.
name: Maloney, Susan E.
name: Dougherty, Joseph D.
Publisher: name: figshare
Year: 2025
Subjects: subject: Genetics
subject: FOS: Biological sciences

description: Supplementary Material 1. Figure 1: Open field assay revealed chromosomal and MYT1L genotype effects on activity. A) MYT1L genotype of all mice across 19 Myt1l+/- XX x Myt1l+/+ XYM litters, including experimental mice used in open field and social operant assays. MYT1L mutation was inherited by significantly fewer offspring than expected by mendelian heritability patterns. B) Sex factor breakdown of all mice across 24 litters, including experimental mice used in open field and social operant assays. The 24 litters included 19 Myt1l Het XX x Myt1l WT XYM litters, and 5 Myt1l WT XX x Myt1l WT XYM litters. Inheritance of modified third chromosome with Sry gene followed expected mendelian heritability. C) Diagram of open field chamber. Dashed red line designates boundary between center zone (orange) and perimeter zone (no color). D) XX mice travel a greater distance in the perimeter than XY mice. E) MYT1L Het mice travel a greater distance in the perimeter than MYT1L WT mice. F) XX mice travel a greater distance in the center than XY mice. G) MYT1L Het mice travel a greater distance in the center than MYT1L WT mice. H) MYT1L Het mice entered the perimeter significantly more than MYT1L WT mice. I) MYT1L Het mice entered the center significantly more than MYT1L WT mice. J) XX mice spend more time in the perimeter zone than XY mice. K) XX mice spend less time in the center zone than XY mice. L) XY mice spend more time on average per visit in the center zone compared to XX mice. In the full univariate model, sex chromosomes and gonads interact to influence mean time per visit in the center (p=0.048). Specifically, XX mice with testes typically spent less time per visit in the center while XY mice with testes spent significantly more time per visit in the center. M) MYT1L Het mice spend less time in the center per visit compared to MYT1L WT mice. For all panels, error bars indicate SEM. Asterisk (*) indicates variables that underwent square root transformation to normalize data distribution. Figure 2: Conditioning achievement did not depend on MYT1L genotype or sex factors. A) Mean of daily nosepokes across the FR1 testing period, with correct nosepokes in blue and incorrect nosepokes in red. Top row are MYT1L WT and bottom row are MYT1L Het B) Histogram showing frequency of achievers (blue) and non-achievers (red) per group. C) FR1 day achievers reached the third consecutive day of criteria across the eight experimental groups. For all panels, error bars indicate SEM. Figure 3: Gonadal and chromosomal sex independently act to alter social seeking and orienting in four core genotypes mice. A) Mean Daily Rewards across all 8 groups. B) Mean Daily Correct Nosepokes across all 8 groups. C) Mean Time in the Interaction Zone across all 8 groups during FR1. D) Mean Daily Attempts across all 8 groups. E) Mean Daily Interaction Time across all groups. F) Mean Daily Interactions across all 8 groups. G) Mean breakpoint across all 8 groups, averaged from 3 separate PR trials. H) Mean Daily Entries into the Interaction Zone across all 8 groups during FR1. I) Mean Daily Entries into the Interaction Zone across all 8 groups during habituation. J) Mean Time in the Interaction Zone across all 8 groups during habituation. K) MYT1L Het test mice enter the interaction zone more often than MYT1L WT mice during FR1. L) Batch was the main driver of variation in Mean Time in the Interaction Zone during habituation, primarily driven by batch 4. For all panels, error bars indicate SEM. Asterisk (*) indicates variables that underwent square root transformation to normalize data distribution. Cross (+) indicates variables that underwent natural log transformation to normalize data distribution. Figure 4: Mega-analysis shows variation in sex bias in social motivation is attributable to cohort effects. A) Mean Daily Rewards for control groups across 7 social operant cohorts, with cohort effect shown. B) Mean Daily Rewards for 7 social operant cohorts split by sex shows no difference in social seeking. C) Mean Time in the Interaction Zone with cohort effect shown. D) Mean Time in the Interaction Zone Rewards for 7 social operant cohorts split by sex shows males spend more time in the interaction zone than females. E) Mean Distance Traveled with cohort effect shown. F) Mean Distance Traveled in the Interaction Zone Rewards for 7 social operant cohorts split sex shows no difference in activity. G) Mean Distance Traveled for each social operant cohort split by sex demonstrates two social operant cohorts (041519 and 102519) show significantly higher activity in females than males. H) Mean Distance Traveled was the only variable to have significant effects with age, namely driven by the 66-86 age group. Statistics indicate results from cohort subset of mega-analysis ANOVA and multiple comparisons (Table S5) and is not the same as reported sex bias from original published experiments (Table S4) which includes all experimental groups. For all panels, error bars indicate SEM. Asterisk (*) indicates variables that underwent square root transformation to normalize data distribution before analysis. Table 1. Table 2. Table 3: Analysis on subset of consistent achievers (Fig.S2B). Table 4: Summary of Cohorts Included in Social Operant Mega-Analysis. Table 5: Mega-Analysis on 7 published social operant cohorts (Fig S3).