RGS6 Mediates Effects of Voluntary Running on Adult Hippocampal Neurogenesis

Abstract

Voluntary running enhances adult hippocampal neurogenesis, with consequences for hippocampal-dependent learning ability and mood regulation. However, the underlying mechanism remains unclear. Here, we show that voluntary running induces unique and dynamic gene expression changes specifically within the adult-born hippocampal neurons, with significant impact on genes involved in neuronal maturation and human diseases. We identify the regulator of G protein signaling 6 (RGS6) as a key factor that mediates running impact on adult-born neurons. RGS6 overexpression mimics the positive effects of voluntary running on morphological and physiological maturation of adult new neurons and reduced sensitivity of adult-born neurons to the inhibitory effect of GABA B (g-Aminobutyric acid B) receptor activation. Knocking down RGS6 abolishes running-enhanced neuronal maturation and hippocampal neurogenesis-dependent learning and anxiolytic effect. Our study provides a data resource showing genome-wide intrinsic molecular changes in adult-born hippocampal neurons that contribute to voluntary running-induced neurogenesis.

sncRiboTag-Seq: Cell-type-specific RiboTag-Seq for cells in low abundance in mouse brain tissue

“…The HA-ribosome bound RNA ( Figures 6 and 7 ) and the next-generation sequencing libraries ( Figure 8 ) produced using our method were high quality. Our sequencing results demonstrated consistency across all samples ( Gao et al., 2020 ) ( Figure 9 ). This method proves powerful in identifying intrinsic genome-wide molecular changes in adult new neurons and can be applied to other small number of cells in mature tissues.…”

“…The numbers of cells needed to obtain this amount of RNA varies depending on cell types. In our study ( Gao et al., 2020 ), we have obtained about 0.8 ng and 4.1 ng RNA from immature and mature adult-born hippocampal neurons of one mouse, respectively ( Figure 7 ). Based on our published papers ( Guo et al., 2011 ; Li et al., 2016 ), we estimated that the number of RiboTag-expressing cells in the hippocampus of one adult Nestin-CreERT2;RiboTag mouse induced by tamoxifen injections should be about 30,000 (7–14 days after tamoxifen injection) and 40,000 (28 days after tamoxifen injection) for immature and mature new neurons, respectively.…”

“…This lysate can be used for isolating Input sample RNA. Add 3 μL of the HA antibody into the rest of the lysate and put on a rotator and rotate for 6 h. CRITICAL: The amount of HA antibody used here is dependent on the number of RiboTag-expressing cells in the tissues of interest, 3 μL HA antibody used here is for induced-RiboTag expression in newborn neurons in two dentate gyri (from one mouse) ( Gao et al., 2020 ). Slightly more HA antibody is fine, however, too much antibody might lead to unspecific IPs.…”

“…We expect that sncRiboTag-Seq protocol will yield high quality and consistent data. In our published paper ( Gao et al., 2020 ), we aimed to study gene expression changes adult new neurons that exist in low numbers in the brain. We crossed RiboTag mice ( Sanz et al., 2009 ) with neural stem cell driver Cre mice ( Nestin-CreER T2 ) ( Lagace et al., 2007 ; Li et al., 2018 ) and performed sncRiboTag-Seq.…”

“…This protocol describes cell-type-specific expression of tagged ribosome and the use of ribosome tagging followed by RNA-seq to identify translatome of low number and sparse cells in mouse brains without disruptive cell isolation. For complete details on the use and execution of this protocol, please refer to Gao et al. (2020) …”