The mammalian brain contains numerous diverse cell types and functional states, which may undergo changes during disease processes. We are developing a three-dimensional proteomic database of the mouse brain captured at cellular resolution to provide fundamental knowledge of cell type distributions and structural morphology in the healthy adult mouse. These proteomic profiles will link new knowledge generated by single cell transcriptomic profiling to functional analyses.   To build this reference, we have established an efficient IHC pipeline to screen monoclonal antibodies (mAbs) for compatibility with whole-mount labeling and imaging based on our advanced iDISCO methods. We are building our reference using monoclonal antibodies because of the advantages these reagents provide in consistent quality and reproducibility. Employing these antibodies, we generate 2D and 3D datasets through whole mount imaging.

Our purpose in this project is three-fold: 1) To provide 3D whole mount proteomic datasets which can directly be used by other scientists to facilitate versatile molecular analyses of specific brain structures. 2) To build a reliable mAb reference list for numerous protein targets, to contribute to knowledge of reliable reagents and reproducibility within the field. 3) To disseminate optimized protocols with reference IHC results to direct the successful utilization of each mAb clone.

By sharing our resulting proteomic datasets on our publicly-accessible online platform (coming soon), we hope our reference can provide a valuable resource for the field.

We study how neural axon guidance establishes properly-formed neural circuitry in the developing mouse embryo. We use both in vivo genetics and in vitro culture models of embryonic neurons to investigate the mechanisms by which extending axons are able to navigate precisely in the complex environment and reach their proper targets over long distances. We are interested in understanding how myriad environmental cues (canonical guidance molecules and other ECM factors) orchestrate the dynamic axonal responses to form diverse neural projections. Studying this process will not only help us to understand the blueprint of adult neural circuitry formation to ensure mature functions, but also provide critical insights for mechanisms underlying later axonal plasticity, pruning and degeneration during aging and disease.