A high-resolution subcellular map of proteins in ciliated cells

Motile cilia are complex structures regulated by thousands of genes, essential for various physiological functions such as respiration and reproduction. Dysfunction can lead to severe conditions like primary ciliary dyskinesia (PCD), underscoring the need for a deeper understanding of the specific ciliary compartments at a molecular level. Here, we developed a stringent high-throughput workflow combining multiplex immunohistochemistry and automated image analysis to investigate protein expression and subcellular localization in motile ciliated cells across five different tissues: nasopharynx, bronchus, fallopian tube, endometrium, and cervix. We characterized proteins with previously unknown roles in motile cilia, including several proteins mapped for the first time. Expression varied across tissues, suggesting motile cilia proteins were not universally expressed among the included tissues. Our presented approach holds promise for mapping the entire motile cilia proteome in health and disease, forming the basis for further understanding of mechanisms underlying ciliopathies.