Single-cell analysis of the nervous system at small and large scales with instant partitions.

Single-cell RNA sequencing is a new frontier across all biology, particularly in neuroscience. While powerful for answering numerous neuroscience questions, limitations in sample input size, and initial capital outlay can exclude some researchers from its application. Here, we tested a recently introduced method for scRNAseq across diverse scales and neuroscience experiments. We benchmarked against a major current scRNAseq technology and found that PIPseq performed similarly, in line with earlier benchmarking data. Across dozens of samples, PIPseq recovered many brain cell types at small and large scales (1,000-100,000 cells/sample) and was able to detect differentially expressed genes in an inflammation paradigm. Similarly, PIPseq could detect expected and new differentially expressed genes in a brain single cell suspension from a knockout mouse model; it could also detect rare, virally-la-belled cells following lentiviral targeting and gene knockdown. Finally, we used PIPseq to investigate gene expression in a nontraditional model species, the little skate (Leucoraja erinacea). In total, PIPSeq was able to detect single-cell gene expression changes across models and species, with an added benefit of large scale capture and sequencing of each sample.

Anticipatory coarticulation and reaction time to phoneme targets in spontaneous speech.

The subject's task was to listen to continuous utterances and to press a reaction-time (RT) button upon hearing an assigned phoneme target. Stimulus materials were utterances of 7--13 syllables in length excised from tape-recorded spontaneous speech; each contained a target located in early, middle or late positions in the utterance. Either 200 msec of silence was tape-spliced into the utterance 33, 67, or 100 msec prior to the target, or the utterance was left intact (as spoken). The main results were faster RT to targets following the silent interval by 33 msec than to targets in the intact version or targets following the silent interval by 10 msec, that is, facilitative effects on target RT varied directly with proximity to target of silent interval. As with previous results using citation-form stimulus materials, these results were interpreted in terms of information from anticipatory coarticulation provided to the subject in advance, permitting extra time to allow the target to be anticipated across the silent interval.

Reaction time to temporally-displaced phoneme targets in continuous speech.

In seven experiments, reaction time (RT) was recorded to phoneme targets in sentences. By means of tape splicing or other experimental interventions preceding the target, speech information was or was not discarded, and targets were either temporally displaced "early" or "late" or remained on time." RT to displaced targets was slower than to on-time targets, except when the tape-splicing manipulation in effect presented coarticulatory target information in advance, in which case RT was faster than RT to on-time targets. The latter manipulation also produced faster RT than RT to the same targets in the original sentence, that is, that containing no experimental intervention at all. In three of the experiments, half the targets were contained in stressed syllables, half in unstressed syllables. Stressed and unstressed targets were affected differently, depending upon practice and other factors. Results were interpreted in terms of listener expectancies based on timing redundancy in continuous speech. They indicate an interaction between the effects of segmental and suprasegmental cues during ongoing perception.