A neural substrate for negative affect dictates female parental behaviour.

# Abstract

Parental behaviours are essential to secure survival and wellbeing of newborns. Concomitantly, parenting also limits negative affective states in adults, which emerge when the coping with neonatal distress becomes a challenge. Whether neural circuits that process negative affect orchestrate components of parental behaviours remains, however, unknown. Here, we identify functional and transcriptional signatures of parental behaviours in neurons of the negative emotion center lateral habenula receiving bed nucleus of the stria terminalis innervation (BNSTLHb). Calcium imaging and optogenetic manipulations reveal that LHb neurons of virgin female mice increase their activity in response to pup distress vocalization and are necessary for pup calls-driven aversive behaviours. Accordingly, LHb activity rises during the retrieval of a pup to a nest, a behaviour worsened by optogenetic LHb inactivation. Intersectional cell identification and transcriptional profiling associates the BNSTLHb neuronal population to parental behaviours and outlines gene expression in female virgins that is similar to mothers but divergent from non-parental virgin male mice. Accordingly, optogenetic activation and inactivation of the BNST-LHb pathway maximizes and suppresses, respectively, the parental behaviour. Finally, tracking and manipulating single BNSTLHb cell activity demonstrates specificity of this neuronal subset for encoding negative affect and pup retrieval, but not sociability amongst conspecifics. Thus, BNSTLHb cells are operational for female parenting, demonstrating that a negative affect neural circuit processes newborn distress signals and limits them through female parenting.

# Files available to generate figures.

- Fastq Files : Raw reads output

Alignment of sequenced reads to the mouse genome (GRCm38) and filtered gene–barcode matrices were realized by running Cell Ranger Single-Cell Software Suite v5.0.1 (10X Genomics).

The cell ranger count function was used to generate filtered gene/cell expression UMI corrected matrices by selecting probable nuclei and removing empty lipid droplets.

Command line use for cell ranger  /opt/cellranger-5.0.1/cellranger count --id=MAc --transcriptome=/refgenome/refdata-gex-mm10-2020-A --libraries=library.csv --expect-cells=2000 --include-introns --localcores=20 --localmem=32

MAc_S1_L001_R1_001.fastq.gz
MAc_S1_L001_R2_001.fastq.gz
MBc_S1_L001_R1_001.fastq.gz
MBc_S1_L001_R2_001.fastq.gz
MCc_S1_L001_R1_001.fastq.gz
MCc_S1_L001_R2_001.fastq.gz

- src.src  R script to reproduce analysis.
- source.src R source script containing useful functions.

- singlecell_countmatrix.tsv.gz: is a count matrix in tab-separated format. It contains the result of the single cell quantification for all genes and all cells after QC. The  values represent the number of exonic reads mapping into each gene after UMI correction. The first columns contain the gene name that is quantified.

- singlecell_metadata.tsv: is a table containing metadata information for each cells analyse in the paper in tab-separated. 

- all_umap.Rds Secondary processed file use in the script.
- habenula.integrated.Rds Secondary processed file use in the script containing external dataset use in this analysis.