Scaled and Efficient Derivation of Loss of Function Alleles in Risk Genes for Neurodevelopmental and Psychiatric Disorders in Human iPSC.

Translating genetic findings for neurodevelopmental and psychiatric disorders (NPD) into actionable disease biology would benefit from large-scale and unbiased functional studies of NPD genes. Leveraging the cytosine base editing (CBE) system, here we developed a pipeline for clonal loss-of-function (LoF) allele mutagenesis in human induced pluripotent stem cells (hiPSCs) by introducing premature stop-codons (iSTOP) that lead to mRNA nonsense-mediated-decay (NMD) or protein truncation. We tested the pipeline for 23 NPD genes on 3 hiPSC lines and achieved highly reproducible, efficient iSTOP editing in 22 NPD genes. Using RNAseq, we confirmed their pluripotency, absence of chromosomal abnormalities, and NMD. Interestingly, for three schizophrenia risk genes (SETD1A, TRIO, CUL1), despite the high efficiency of base editing, we only obtained heterozygous LoF alleles, suggesting their essential roles for cell growth. We replicated the reported neural phenotypes of SHANK3-haploinsufficiency and found CUL1-LoF reduced neurite branches and synaptic puncta density. This iSTOP pipeline enables a scaled and efficient LoF mutagenesis of NPD genes, yielding an invaluable shareable resource.

Evaluation of smartphone interactions on drivers' brain function and vehicle control in an immersive simulated environment.

Smartphones and other modern technologies have introduced multiple new forms of distraction that color the modern driving experience. While many smartphone functions aim to improve driving by providing the driver with real-time navigation and traffic updates, others, such as texting, are not compatible with driving and are often the cause of accidents. Because both functions elicit driver attention, an outstanding question is the degree to which drivers' naturalistic interactions with navigation and texting applications differ in regard to brain and behavioral indices of distracted driving. Here, we employed functional near-infrared spectroscopy to examine the cortical activity that occurs under parametrically increasing levels of smartphone distraction during naturalistic driving. Our results highlight a significant increase in bilateral prefrontal and parietal cortical activity that occurs in response to increasingly greater levels of smartphone distraction that, in turn, predicts changes in common indices of vehicle control.

Head and eye movements in rats with pontine reticular lesions in comparison with primates: a scientific memoir and a fresh look at some old and 'new' data.

The author recounts the process of discovery in Philip Teitelbaum's laboratory, which began with the observation of vestibular head stabilization in a rat with brainstem lesions, of the essential roles of the pontine reticular formation (PRF) in the rat in ipsiversive head as well as eye movements. The PRF in the rat appears to be in the pathways for most direction-changing movements of the eyes and head, leaving vestibular and optokinetic stabilizing movements intact and uninterrupted. The author postulates that a response to the sliding of feet or paws, or a "substrate-kinetic reflex," works together with vestibular and optokinetic reactions to stabilize an animal's directions of gaze and locomotion on the ground. Previously unpublished data are presented from later observations and recordings of rats with kainic acid lesions in the PRF, which support the conclusion that neurons in the PRF are essential for head as well as eye movements in the rat. In contrast, Volker Henn observed no obvious loss of head movements in monkeys that had a loss of fast eye movements from kainic acid lesions of the PRF. The author and others observed that quick phases of head nystagmus develop some time after quick phases of ocular nystagmus in normal human infants; in other words, after the PRF is functioning for eye movements. The author concludes that in primates, the pathway for head movements through the PRF has been replaced by a newer pathway, leaving certain PRF regions to be devoted to mediating only eye movements.