Induced Pluripotent Stem Cells with Six Reprogramming Factors from Prairie Vole, Which is an Animal Model for Social Behaviors

Abstract

Prairie voles show strong pair bonding with their mating partners, and they demonstrate parental behavior toward their infants, indicating that the prairie vole is a unique animal model for analysis of molecular mechanisms of social behavior. Until a recent study, the signaling pathway of oxytocin was thought to be critical for the social behavior of prairie voles, but neuron-specific functional research may be necessary to identify the molecular mechanisms of social behavior. Prairie vole pluripotent stem cells of high quality are essential to elucidate the molecular mechanisms of social behaviors. Generation of high-quality induced pluripotent stem cells (iPSCs) would help to establish a genetically modified prairie vole, including knockout and knock-in models, based on the pluripotency of iPSCs. Thus, we attempted to establish high-quality prairie vole-derived iPSCs (pv-iPSCs) in this study. We constructed a polycistronic reprogramming vector, which included six reprograming factors (Oct3/4, Sox2, Klf4, c-myc, Lin28, and Nanog). Furthermore, we evaluated the effect of six reprogramming factors, which included Oct3/4 with the transactivation domain (TAD) of MyoD. Implantation of the pv-iPSCs into immunodeficient mice caused a teratoma with three germ layers. Furthermore, the established pv-iPSCs tested positive for stem cell markers, including alkaline phosphatase activity (ALP), stage-specific embryonic antigen (SSEA)-1, and dependence on leukemia inhibitory factor (LIF). Our data indicate that our newly established pv-iPSCs may be a useful tool for genetic analysis of social behavior.

Immortalized prairie vole-derived fibroblasts (VMF-K4DTs) can be transformed into pluripotent stem cells and provide a useful tool with which to determine optimal reprogramming conditions

“…In both cases, the expression of the IRES-GFP locus, located downstream from the reprogramming factor cassette, was lower after sequential passage than in primary colonies; this decrease could potentially be explained by the silencing of the exogenous promoter. The same phenomenon was observed in later passages of iPSCs derived from primary prairie vole cells [2], indicating that immortalized cells had retained the original characteristics of the primary prairie vole cells from which they were derived.…”

“…Previously, we reported the establishment of iPSCs from embryonic prairie vole-derived primary cells [2], but this was only made possible by the extensive optimization of experimental conditions using immortalized adult prairie vole cells described in the present study. The prairie vole is a useful animal model with which to analyze social behaviors such as pair bonding [17, 18].…”

“…These data were not included in our previous article [2]. The cells used here were immortalized by the expression of mutated CDK4, cyclin D, and TERT.…”

“…Scale bar = 100 μm., respectively. A detailed description of the composition of the medium used has been reported previously [2]. …”

“…Previously, we showed that embryonic cells from the prairie vole ( Microtus ochrogaster ) require the expression of six reprogramming factors to efficiently transform into iPSCs [2]. In the present study, we present the detailed experimental data describing the optimization of the prairie vole iPSC protocol reported previously.…”