Available methods for differentiating human embryonic (ES) and induced pluripotent stem (iPS) cells into neurons are often cumbersome, slow and variable. Alternatively, human fibroblasts can be directly converted into induced neuronal (iN) cells. However, with present techniques conversion is inefficient, synapse formation is limited, and only small amounts of neurons can be generated. Here, we show that human ES and iPS cells can be converted into functional iN cells with nearly 100% yield and purity in less than two weeks by forced expression of a single transcription factor. The resulting ES-iN or iPS-iN cells exhibit quantitatively reproducible properties independent of the cell line of origin, form mature pre- and postsynaptic specializations, and integrate into existing synaptic networks when transplanted into mouse brain. As illustrated by selected examples, our approach enables large-scale studies of human neurons for questions such as analyses of human diseases, examination of human-specific genes, and drug screening.

Increased fear memory generalization is associated with post-traumatic stress disorder, but the circuit mechanisms that regulate memory specificity remain unclear. Here, we define a neural circuit, composed of the medial prefrontal cortex, the N. reuniens, and the hippocampus, that controls fear memory generalization. Inactivation of prefrontal inputs into the N. reuniens or direct silencing of N. reuniens projections enhanced fear memory generalization, whereas constitutive activation of N. reuniens neurons decreased memory generalization. Direct optogenetic activation of phasic and tonic action-potential firing of N. reuniens neurons during memory acquisition enhanced or reduced memory generalization, respectively. We propose that the N. reuniens determines the specificity and generalization of memory attributes for a particular context by processing information from the medial prefrontal cortex en route to the hippocampus.

In forebrain neurons, knockout of synaptotagmin-1 blocks fast Ca2+-triggered synchronous neurotransmitter release, but enables manifestation of slow Ca2+-triggered asynchronous release. Here, we show using single-cell PCR that individual hippocampal neurons abundantly co-express two Ca2+-binding synaptotagmin isoforms, synaptotagmin-1 and synaptotagmin-7. In synaptotagmin-1 deficient synapses of excitatory and inhibitory neurons, loss-of-function of synaptotagmin-7 suppressed asynchronous release. This phenotype was rescued by wild-type but not mutant synaptotagmin-7 lacking functional Ca2+-binding sites. Even in synaptotagmin-1 containing neurons, synaptotagmin-7 ablation partly impaired asynchronous release induced by extended high-frequency stimulus trains. Synaptotagmins bind Ca2+ via two C2-domains, the C2A- and C2B-domains. Surprisingly, synaptotagmin-7 function selectively required its C2A-domain Ca2+-binding sites, whereas synaptotagmin-1 function required its C2B-domain Ca2+-binding sites. Our data show that nearly all Ca2+-triggered release at a synapse is due to synaptotagmins, with synaptotagmin-7 mediating a slower form of Ca2+-triggered release that is normally occluded by faster synaptotagmin-1-induced release, but becomes manifest upon synaptotagmin-1 deletion.