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Unbiased classification of mosquito blood cells by single-cell genomics and high-content imaging

By Maiara S. Severo, Jonathan J.M. Landry, Randall L. Lindquist, Christian Goosmann, Volker Brinkmann, Paul Collier, Anja E Hauser, Vladimir Benes, Johan Henriksson, Sarah A Teichmann, Elena A. Levashina

Posted 15 Dec 2017
bioRxiv DOI: 10.1101/234492 (published DOI: 10.1073/pnas.1803062115)

Mosquito blood cells are ancestral immune cells that help control infection by vector-borne pathogens. Despite their importance, little is known about mosquito blood cell biology beyond the ambiguous morphological and functional criteria used for their classification. Here we combined the power of single-cell RNA-sequencing, high-content imaging flow cytometry and single-molecule RNA hybridization to analyze blood cells of the malaria mosquito Anopheles gambiae. By demonstrating that blood cells express nearly half of the mosquito transcriptome, our dataset represents an unprecedented view into their transcriptional machinery. Analyses of differentially expressed genes identified transcriptional signatures of two distinct cell types that challenge the current morphology-based classification of these cells. We further demonstrated an active transfer of a cellular marker between blood cells that confounds their identity. We propose that cell-to-cell exchange is broadly relevant for cell type classification and may account for the remarkable cellular diversity observed in nature.

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