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Cellular membrane asymmetry is a hallmark characteristic of all eukaryotic cells. The balance of phospholipid composition within the cytoplasmic inner leaflet and the extracellular outer leaflet of the plasma membrane (PM) maintains cellular function and vitality. The proper exposure of particular phospholipids is necessary to maintain cellular signalling, controlled apoptosis, and vesicle transportation among other roles. Phospholipid asymmetry is coordinated by P4-type phospholipid transferases (flippases or ATPases). ATP11A, ATP11B, and ATP11C belong to class VI of the P4-flippase family (vertebrates) and are responsible for the movement of phosphatidylserine (PS) from the outer leaflet to the inner leaflet of the PM. To date, there is a lack of knowledge of the tissue specific expression of these three flippases on a whole-organism level in a vertebrate system. Here we have determined the spatial-temporal expression profiles of each gene in a zebrafish model using in situ hybridization and performed comparative phylogenetic analyses with other vertebrates. Our data reveals sequence similarity between vertebrate flippases and specific synteny of zebrafish and human chromosomes. Both atp11b and atp11c are maternally expressed in zebrafish, while zygotic expression analysis demonstrates tissue and temporal specificity for all three genes. atp11a is expressed in the neural crest cells as well as in the developing eye and ear, while atp11b is expressed early in the ventricular epithelial lining and later in the ear. atp11c is expressed in the anterior most rhombomeres of the hindbrain, pharyngeal arches, and liver. Our expression data suggests that each of the three flippases are integral for the development of specific tissues, and aberrant function of either could lead to visual, hearing, neural, or liver dysfunction., Competing Interests: Declaration of competing interest There are no conflicts of interest to declare., (Copyright © 2020 Elsevier B.V. All rights reserved.)