Journal: bioRxiv

Article Title: Sex-specific regulation of social play in juvenile rats by oxytocin neurons in the hypothalamus and oxytocin signaling in the nucleus accumbens

doi: 10.1101/2025.11.11.687699

Figure Lengend Snippet: (A) Timeline of experimental procedures in which juvenile rats (26-day-old) underwent stereotaxic surgery for infusion of AAV-OXTp-hM3Dq-mCherry into the PVN. Following recovery and habituation to IP injections, all rats underwent two days of social play testing in which either saline or CNO was administered 30 min prior to testing. Drug conditions were randomly assigned and counterbalanced over the two days of testing. (B) Rat brain atlas image (modified from Paxinos and Watson, 2007) indicating the location and representative image of mCherry expression within PVN OXT neurons. (C-F) Magnifications of image in B denoting mCherry (C, red) and OXT (D, green) positive neurons that are co-localized (F, yellow) using DAPI (E) as a cellular marker. (G-L) Administration of CNO significantly decreased the duration of social play in males and had a strong tendency towards increasing the duration of social play in females (G) as well as significantly decreased the number of nape attacks in males (H). CNO administration did not alter number of pins (I), duration of social investigation (J), duration of allogrooming (K), or total duration of social behaviors (L). Social play, investigation, allogrooming and combined social behaviors are expressed as a percentage of total time. 2-way ANOVA, Sex x Drug interaction; *: p < 0.05, **: p < 0.01; Holm’s-Sidak post-hoc tests.

Article Snippet: Slides were then rinsed in wash buffer and coverslipped with Vectashield hardset antifade mounting medium with a DAPI counterstain (H-1500-10, Vector Laboratories) and stored at 4°C.

Techniques: Saline, Modification, Expressing, Marker

Journal: bioRxiv

Article Title: Sex-specific regulation of social play in juvenile rats by oxytocin neurons in the hypothalamus and oxytocin signaling in the nucleus accumbens

doi: 10.1101/2025.11.11.687699

Figure Lengend Snippet: (A) Timeline of experimental procedures in which juvenile rats (26-day-old) underwent stereotaxic surgery for infusion of AAV-OXTp-hM3Dq-mCherry into the SON. Following recovery and habituation to IP injections, all rats underwent two days of social play testing in which either saline or CNO was administered 30 min prior to testing. Drug conditions were randomly assigned and counterbalanced over the two days of testing. (B) Rat brain atlas image (modified from Paxinos and Watson, 2007) indicating the location of the SON and representative image of AAV-OXTp-hM3Dq-mCherry expression in OXT neurons of the SON as well as magnification of image denoting OXT (C, green) and mCherry (D, red) positive neurons that are co-localized using DAPI (E, blue) as a cellular marker. Irrespective of drug treatment, females showed lower duration of social play (G) and fewer nape attacks than males (H). CNO administration decreased the number of pins (I) and increased the duration of social investigation (J) in both sexes compared to saline administration. Neither drug nor sex altered the duration of allogrooming (K) and total duration of social behaviors (L). Durations of social play (G), social investigation (J), allogrooming (K), and total social behavior (L) are expressed as a percentage of total time. 2-way ANOVA main effect of drug, *: p < 0.05; main effect of sex, ##: p < 0.01; ###: p < 0.001.

Article Snippet: Slides were then rinsed in wash buffer and coverslipped with Vectashield hardset antifade mounting medium with a DAPI counterstain (H-1500-10, Vector Laboratories) and stored at 4°C.

Techniques: Saline, Modification, Expressing, Marker

Journal: bioRxiv

Article Title: Sex-specific regulation of social play in juvenile rats by oxytocin neurons in the hypothalamus and oxytocin signaling in the nucleus accumbens

doi: 10.1101/2025.11.11.687699

Figure Lengend Snippet: Exposure to social play does not alter activation patterns within the NAc in juvenile male and female rats but is negatively correlated with the number of activated oxtr mRNA+ neurons and the proportion of activated oxtr mRNA+ neurons in the anterior NAc (A) Representative image of oxtr and fos mRNA expressing nuclei in the NAc. (B-E) Examples of labeled nuclei from the white-boxed region of (A) positive for fos (B; green puncta), oxtr (C; magenta puncta), DAPI (D; blue nuclear stain), and double-labeled nuclei (E). Neither play condition nor sex altered the (F) average number of fos mRNA+ nuclei, (G) average number of oxtr+fos mRNA double-labeled nuclei, (H) the proportion of activated oxtr mRNA+ nuclei, or (I) the proportion of activated nuclei expressing oxtr mRNA. 2-way ANOVA; p > 0.05 for all. (F’) There is a significant negative correlation between duration of social play and the average number of fos + nuclei, as well as (I’) duration of social play and the percentage of activated oxtr+ nuclei. The association between social play and fos+ nuclei was not detected in either males (F’’) or females (F’’’) alone, but there was a significant negative correlation between social play and the proportion of activated oxtr+ nuclei in males only (I’’). No association between social play duration and average oxtr+fos+ nuclei (G’-G’’’) or proportion of activated nuclei co-expressing oxtr (H’-H’’’) was detected.

Article Snippet: Slides were then rinsed in wash buffer and coverslipped with Vectashield hardset antifade mounting medium with a DAPI counterstain (H-1500-10, Vector Laboratories) and stored at 4°C.

Techniques: Activation Assay, Expressing, Labeling, Staining