Characterization of transcriptional profiles associated with stress-induced neuronal activation in Arc-GFP mice.

Chronic stress has become a predominant factor associated with a variety of psychiatric disorders, such as depression and anxiety, in both human and animal models. Although multiple studies have looked at transcriptional changes after social defeat stress, these studies primarily focus on bulk tissues, which might dilute important molecular signatures of social interaction in activated cells. In this study, we employed the Arc-GFP mouse model in conjunction with chronic social defeat (CSD) to selectively isolate activated nuclei (AN) populations in the ventral hippocampus (vHIP) and prefrontal cortex (PFC) of resilient and susceptible animals. Nuclear RNA-seq of susceptible vs. resilient populations revealed distinct transcriptional profiles linked predominantly with neuronal and synaptic regulation mechanisms. In the vHIP, susceptible AN exhibited increased expression of genes related to the cytoskeleton and synaptic organization. At the same time, resilient AN showed upregulation of cell adhesion genes and differential expression of major glutamatergic subunits. In the PFC, susceptible mice exhibited upregulation of synaptotagmins and immediate early genes (IEGs), suggesting a potentially over-amplified neuronal activity state. Our findings provide a novel view of stress-exposed neuronal activation and the molecular response mechanisms in stress-susceptible vs. resilient animals, which may have important implications for understanding mental resilience.

INTACT vs. FANS for Cell-Type-Specific Nuclei Sorting: A Comprehensive Qualitative and Quantitative Comparison.

Increasing numbers of studies seek to characterize the different cellular sub-populations present in mammalian tissues. The techniques "Isolation of Nuclei Tagged in Specific Cell Types" (INTACT) or "Fluorescence-Activated Nuclei Sorting" (FANS) are frequently used for isolating nuclei of specific cellular subtypes. These nuclei are then used for molecular characterization of the cellular sub-populations. Despite the increasing popularity of both techniques, little is known about their isolation efficiency, advantages, and disadvantages or downstream molecular effects. In our study, we compared the physical and molecular attributes of sfGFP+ nuclei isolated by the two methods-INTACT and FANS-from the neocortices of Arc-CreERT2 × CAG-Sun1/sfGFP animals. We identified differences in efficiency of sfGFP+ nuclei isolation, nuclear size as well as transcriptional (RNA-seq) and chromatin accessibility (ATAC-seq) states. Therefore, our study presents a comprehensive comparison between the two widely used nuclei sorting techniques, identifying the advantages and disadvantages for both INTACT and FANS. Our conclusions are summarized in a table to guide researchers in selecting the most suitable methodology for their individual experimental design.

Selective targeting of chronic social stress-induced activated neurons identifies neurogenesis-related genes to be associated with resilience in female mice.

Prolonged social stress is a major cause for depression in humans and is associated with a wide range of subsequent pathophysiological changes such as elevated blood pressure. A routinely used model for investigating this kind of stress in mice is the chronic social defeat paradigm where a smaller intruder is exposed to an aggressive inhabitant of a home cage. This model is restricted to males and includes a high proportion of physical stress that might e.g., interfere with immunological aspects of the stress. The prevalence of depression in humans is even higher in women than in men. Therefore, expanding models to female individuals is desirable. We here tested the social instability model as a tool for administering chronic social stress to female C57BL/6J mice and analyzed short-term as well as long-lasting effects. Animals were housed in groups of four and were shuffled two times a week, resulting in a permanent re-structuration of their social hierarchy. While directly after the stress exposure, serum corticosterone was elevated, increased body weight and fat deposits were observed in stressed mice even one year after discontinuation of the stress. At the behavioral level, animals could be stratified into resilient and susceptible animals directly post-stress, but those subgroups were not distinguishable any more in the long-term analysis. To identify molecular contributors to resilience in the here presented social instability induced stress model, Arc-activity dependent trapping of neurons was conducted in Arc-creERT2/sun1sfGFP mice. RNA samples derived from activated nuclei from the ventral hippocampus, a brain region involved in stress-regulation during attacks or explorative behavior of mice, were subjected to a neurogenesis pathway array. While several genes were differentially regulated by stress, in particular, artemin, a neurotrophic factor was upregulated in resilient versus susceptible individuals.