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Unified single-cell analysis of testis gene regulation and pathology in 5 mouse strains

By Min Jung, Daniel Wells, Jannette Rusch, Suhaira Ahmed, Jonathan Marchini, Simon Myers, Donald F. Conrad

Posted 16 Aug 2018
bioRxiv DOI: 10.1101/393769 (published DOI: 10.7554/eLife.43966)

By removing the confounding factor of cellular heterogeneity, single cell genomics can revolutionize the study of development and disease, but methods are needed to simplify comparison among individuals. To develop such a framework, we assayed the transcriptome in 62,600 single cells from the testes of wildtype mice, and mice with gonadal defects due to disruption of the genes Mlh3, Hormad1, Cul4a or Cnp. The resulting expression atlas of distinct cell clusters revealed novel markers and new insights into testis gene regulation. By jointly analysing mutant and wildtype cells using a model-based factor analysis method, SDA, we decomposed our data into 46 components that identify novel meiotic gene regulatory programmes, mutant-specific pathological processes, and technical effects. Moreover, we identify, de novo, DNA sequence motifs associated with each component, and show that SDA can be used to impute expression values from single cell data. Analysis of SDA components also led us to identify a rare population of macrophages within the seminiferous tubules of Mlh3-/- and Hormad1-/- testes, an area typically associated with immune privilege.We provide a web application to enable interactive exploration of test is gene expression and components at http://www.stats.ox.ac.uk/~wells/testisAtlas.html.

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