The naïve pluripotent state has been shown in mice to lead to broad and more robust developmental potential relative to primed mouse epiblast cells. The human naïve ES cell state has eluded derivation without the use of transgenes, and forced expression of OCT4, KLF4, and KLF2 allows maintenance of human cells in a naïve state [Hanna J, et al. (2010) Proc Natl Acad Sci USA 107 (20):9222-9227]. We describe two routes to generate nontransgenic naïve human ES cells (hESCs). The first is by reverse toggling of preexisting primed hESC lines by preculture in the histone deacetylase inhibitors butyrate and suberoylanilide hydroxamic acid, followed by culture in MEK/ERK and GSK3 inhibitors (2i) with FGF2. The second route is by direct derivation from a human embryo in 2i with FGF2. We show that human naïve cells meet mouse criteria for the naïve state by growth characteristics, antibody labeling profile, gene expression, X-inactivation profile, mitochondrial morphology, microRNA profile and development in the context of teratomas. hESCs can exist in a naïve state without the need for transgenes. Direct derivation is an elusive, but attainable, process, leading to cells at the earliest stage of in vitro pluripotency described for humans. Reverse toggling of primed cells to naïve is efficient and reproducible.

Both the medial prefrontal cortex (mPFC) and hippocampus are implicated in working memory tasks in rodents. Specifically, it has been hypothesized that the mPFC is primarily engaged in the temporary storage and processing of information lasting from a subsecond to several seconds, while the hippocampal function becomes more critical as the working memory demand extends into longer temporal scales. Although these structures may be engaged in a temporally separable manner, the extent of their contributions in the "informational content" of working memory remains unclear. To investigate this issue, the mPFC and dorsal hippocampus (dHPC) were temporarily inactivated via targeted infusions of the GABA A receptor agonist muscimol in rats prior to their performance on a delayed alternation task (DAT), employing an automated figure-eight maze that required the animals to make alternating arm choice responses after 3-, 30-, and 60-sec delays for water reward. We report that inactivation of either the mPFC or dHPC significantly reduced DAT at all delay intervals tested. However, there were key qualitative differences in the behavioral effects. Specifically, mPFC inactivation selectively impaired working memory (i.e., arm choice accuracy) without altering reference memory (i.e., the maze task rule) and arm choice response latencies. In contrast, dHPC inactivation increased both reference memory errors and arm choice response latencies. Moreover, dHPC, but not mPFC, inactivation increased the incidence of successive working memory errors. These results suggest that while both the mPFC and hippocampus are necessarily involved in DAT, they seem to process different informational components associated with the memory task.Working memory is generally defined as cognitive entities (or "central executive" mechanisms) relating to temporary storage and operation of information in both humans and animals (Baddeley and Hitch 1974;Goldman-Rakic 1996;Fuster 2001;Dudchenko 2004). The memory may be about simple sensory stimulus, relatively complex objects, or spatial location (Olton et al. 1979;Delatour and Gisquet-Verrier 1996;Floresco et al. 1997;Hampson et al. 1999). Evidence from primate studies originally implicated the prefrontal cortex (PFC) as being crucial for working memory (Baddeley 1986;Goldman-Rakic 1987;Miller et al. 1991). For instance, damage to the PFC produces impairments in various working memory tasks in humans and non-human primates (Kolb 1990;Fuster 1997;Stuss and Alexander 2000). Additionally, recording and brain imaging studies found neural activity correlates of working memory, i.e., increased PFC activity during the delay period (Fuster and Alexander 1971;Kubota and Niki 1971;Funahashi et al. 1989).In rodents, a delayed alternation task (DAT; employing T, radial-arm, and figure-eight mazes) has been widely used to further investigate the PFC-working memory hypothesis (Murphy et al. 1996;Zahrt et al. 1997;Baeg et al. 2003Baeg et al. , 2007Schoenbaum et al. 2003;Birnbaum et al. 2004;Clinton et al. 2006). In this t...

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