title: Quantitative single cell analysis of cell population dynamics during submandibular salivary gland development and differentiation.
abstract: Epithelial organ morphogenesis involves reciprocal interactions between epithelial and mesenchymal cell types to balance progenitor cell retention and expansion with cell differentiation for evolution of tissue architecture. Underlying submandibular salivary gland branching morphogenesis is the regulated proliferation and differentiation of perhaps several progenitor cell populations, which have not been characterized throughout development, and yet are critical for understanding organ development, regeneration, and disease. Here we applied a serial multiplexed fluorescent immunohistochemistry technology to map the progressive refinement of the epithelial and mesenchymal cell populations throughout development from embryonic day 14 through postnatal day 20. Using computational single cell analysis methods, we simultaneously mapped the evolving temporal and spatial location of epithelial cells expressing subsets of differentiation and progenitor markers throughout salivary gland development. We mapped epithelial cell differentiation markers, including aquaporin 5, PSP, SABPA, and mucin 10 (acinar cells); cytokeratin 7 (ductal cells); and smooth muscle α-actin (myoepithelial cells) and epithelial progenitor cell markers, cytokeratin 5 and c-kit. We used pairwise correlation and visual mapping of the cells in multiplexed images to quantify the number of single- and double-positive cells expressing these differentiation and progenitor markers at each developmental stage. We identified smooth muscle α-actin as a putative early myoepithelial progenitor marker that is expressed in cytokeratin 5-negative cells. Additionally, our results reveal dynamic expansion and redistributions of c-kit- and K5-positive progenitor cell populations throughout development and in postnatal glands. The data suggest that there are temporally and spatially discreet progenitor populations that contribute to salivary gland development and homeostasis.
authors.affiliation: Department of Biological Sciences, University at Albany, State University of New York , 1400 Washington Avenue, Albany, NY 12222 , USA.
authors.forename: Deirdre A
authors.initials: DA
authors.lastname: Nelson
authors.forename: Charles
authors.initials: C
authors.lastname: Manhardt
authors.forename: Vidya
authors.initials: V
authors.lastname: Kamath
authors.forename: Yunxia
authors.initials: Y
authors.lastname: Sui
authors.forename: Alberto
authors.initials: A
authors.lastname: Santamaria-Pang
authors.forename: Ali
authors.initials: A
authors.lastname: Can
authors.forename: Musodiq
authors.initials: M
authors.lastname: Bello
authors.forename: Alex
authors.initials: A
authors.lastname: Corwin
authors.forename: Sean R
authors.initials: SR
authors.lastname: Dinn
authors.forename: Michael
authors.initials: M
authors.lastname: Lazare
authors.forename: Elise M
authors.initials: EM
authors.lastname: Gervais
authors.forename: Sharon J
authors.initials: SJ
authors.lastname: Sequeira
authors.forename: Sarah B
authors.initials: SB
authors.lastname: Peters
authors.forename: Fiona
authors.initials: F
authors.lastname: Ginty
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Gerdes
authors.forename: Melinda
authors.initials: M
authors.lastname: Larsen
keywords: Development; Multiplex analysis; Progenitor cells; Salivary gland