Title: In vivo characterization of adult hippocampal neural stem cell behavior
Abstract: Adult neural stem cells (NSC) are primarily quiescent, in reversible G0 of the cell cycle, and lose their ability to exit quiescence with aging and disease. We have previously found in vitro that the intermediate filament vimentin collapses around aggresome, forming a vimentin cage, and bringing with it bound proteins, such as chaperones and proteasomes. As the NSC divides, it asymmetrically segregates these cellular cargoes to one daughter cell, leading to a decrease in its proliferation rate, while the other daughter remains able to rejuvenate the niche. Although the role of vimentin in quiescence exit has been shown in vitro, its function in a physiological setting in vivo remains unknown. To study how vimentin protein is regulated in vivo, we developed a transgenic mouse line with vimentin tagged the fluorophore. First, we asked what cell types are labelled with vimentin. To answer this question and benchmark this line with current NSC reporter lines, we crossed the Vimentin-mScarlet mouse line with either Nestin-GFP or GFAP-GFP mice and performed RNA sequencing on purified hippocampal cells, identifying unique vimentin-mScarlet populations. To further characterize the cell behaviors of mScarlet+ and/or GFP+ cells, we sorted the purified hippocampal cells and compared their ability to form neurospheres. We observed that a larger fraction of GFP+ cells formed neurospheres than mScarlet+ cells, suggesting that vimentin is not a NSC specific marker. Further, we also performed timelapse analyses of these purified populations to observe NSC behaviors such as time to first division and lineage progression. We found that NSC upregulate vimentin protein as they activate from a primed state. In summary, these studies will provide a stem cell-focused characterization of a novel vimentin-mScarlet mouse line, providing a new resource to the scientific community, and reveal the role of vimentin in NSC dynamics in the adult brain.
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