Wyoming Sensory Biology COBRE

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The objective of this administrative supplement is to enhance our research capabilities in the transcriptomic characterization of the nervous system, and forge a more synergistic interaction among SBC investigators by promoting transcriptomic research and collaborations. These funds will be used to acquire a NextSeq550 Sequencing System (Illumina) that will significantly enhance SBC’s access to transcriptomic research, and strengthen the services provided by the Molecular Analysis Unit (MAU) of the Integrated Microscopy Core (IMCore). Over the past 4-year funding period, the IMCore has made significant advances towards assisting SBC project leaders with their research involving transcriptomics. Currently, there have already been single cell RNAseq and Chipseq experiments that have been conducted in the core, as evidenced in our recent publications. However, these experiments were only able to be carried out through shipping out samples to multiple third- party service centers and core facilities (e.g., Genewiz.com 10x genomics and Harvard Bauer Core Facilities). Although this approach has “sufficed”, several major issues hinder its application among SBC researchers: 1) the shipped samples require -80 degree Fahrenheit/Celsius storage, and transporting them in dry ice adds unnecessary variability to the sample qualities; 2) the quality controls of library preparations are highly variable as they are conducted by different commercial vendors that use different protocols; 3) lack of hands-on technical support inhibits application by new project leaders. Therefore, the purchase of the NextSeq550 will be a game changer for transcriptomic research at the University of Wyoming. We anticipate that the number of projects using single cell RNAseq or the Chipseq approach will increase significantly given the availability of the NextSeq550. The purchase of NextSeq550 is fully justifiable, reasonable, and allocable to grant projects – we currently have four new research projects that outline utilizing this instrument. Project 1: to identify state- dependent changes in neuronal transcription at the level of individual cells. Project 2: single-nucleus RNA-seq will be used to explore transcriptional heterogeneity in the medial prefrontal cortex with differential projections during stress-induced depression. Project 3: to investigate gene expression within reward-specific ensembles – defined as a discrete number of neurons that become synchronously activated – and how that differs in neurons that become activated during the seeking of different types of rewards. Project 4: to answer the following questions: (1) which genes are activated/repressed after injury and during regeneration, and (2) are the transcriptional profiles generated different based on injury mechanism and/or cell type. We anticipate that this administrative supplement will significantly help increase our access to transcriptomic research and collaboration, increase the number of projects that would utilize this instrument during the remainder of the funding period, and help build a strong foundation for Phase II applications.