Neural Circuits and Brainstem Control of LUT Function
project - Identifying distinct neuronal cell types in the brainstem micturition control centers
Our and previous studies have shown voiding function to be mediated by Barrington’s nucleus (Bar) and periaqueductal gray (PAG) regions of the brain. We use RNA sequencing techniques to profile gene expression in single cells or single nuclei from the brainstem region where Barrington’s nucleus is located, as well as from locations upstream and downstream of Barrington’s nucleus. The resulting RNA expression profiles will identify the neuron subgroups in these crucial regions.
Our goal is to study the neuron subpopulations most likely to control the bladder. Transgenic models that selectively express Cre-recombinase permit us to define the subtypes of neurons with roles in normal voiding function and with disorders of bladder function.
We collaborate with the Zeidel and Tsai labs at BIDMC and the BNORC Functional Genomics and Bioinformatics core and use state of the art genomic applications. These studies will provide a clearer picture of the heterogeneity of cells and their genetic basis. In particular, we use next generation techniques and sequencing platforms like Dropseq (Dropseq with nuclei) and sNuc-seq to study the biological, genetic and neural circuits underlying this crucial somatic function.
>> This project aims to create transcriptomic atlases of Barrington’s nucleus in the brainstem and the periaqueductal gray (PAG), and to map the connections between them. <<
- Funding and collaboration on this study: NIH/NIDDK, P20 DK119798 (Zeidel - PI), 2018-2020
Pictured above is the clustering analysis of ~61,000 single cells clustered based on their gene expression profile. The resulting UMAP plot contains 13,900 neurons colored per density clustering, and annotated according to known glutamatergic neuron subtypes in/near Barrington’s nucleus..
Violin plots showing mean +/- S.E.M. of unique molecular identifiers (UMIs) per cluster. The mean +/- S.E.M. genes detected for each cluster and mitochondrial gene percentage.
Dendrogram showing relatedness of glutamatergic neuronal populations in/near Barrington’s nucleus.
Useful references on RNA sequencing techniques:
1) Ecker et al., (2017) Neuron 96, 542-557. 2) Poulin et al., (2016) Nat Neurosci. 19, 1131-1141. 3) Bakken et al., (2018) PLOS-One 13, e0209648. 4) Tsai et al., (2022) Nat.Neurosci. 25, p659-674.
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Links
* DropSeq
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