Generation and characterization of DSPP‐Cerulean/DMP1‐Cherry reporter mice

Abstract

To gain a better understanding of the progression of progenitor cells in the odontoblast lineage, we have examined and characterized the expression of a series of GFP reporters during odontoblast differentiation. However, previously reported GFP reporters (pOBCol2.3-GFP, pOBCol3.6-GFP, and DMP1-GFP), similar to the endogenous proteins, are also expressed by bone-forming cells, which made it difficult to delineate the two cell types in various in vivo and in vitro studies. To overcome these difficulties we generated DSPP-Cerulean/DMP1-Cherry transgenic mice using a bacterial recombination strategy with the mouse BAC clone RP24-258g7. We have analyzed the temporal and spatial expression of both transgenes in tooth and bone in vivo and in vitro. This transgenic animal enabled us to visualize the interactions between odontoblasts and surrounding tissues including dental pulp, ameloblasts and cementoblasts. Our studies showed that DMP1-Cherry, similar to Dmp1, was expressed in functional and fully differentiated odontoblasts as well as osteoblasts, osteocytes and cementoblasts. Expression of DSPP-Cerulean transgene was limited to functional and fully differentiated odontoblasts and correlated with the expression of Dspp. This transgenic animal can help in the identification and isolation of odontoblasts at later stages of differentiation and help in better understanding of developmental disorders in dentin and odontoblasts. K E Y W O R D Sbone, dentin matrix protein 1, dentin sialophosphoprotein, fluorescent protein reporters, Odontoblasts

Dental cell type atlas reveals stem and differentiated cell types in mouse and human teeth

“…A small fraction of the dividing cells showed activation of population-specific transcriptional biases, including an odontoblastic program. Obtaining cells of odontoblast sublineage became possible because we enriched for it by using Dspp cerulean /Dmp1 Cherry transgenic animals 38 . The immunohistochemistry confirmed activation of early odontoblast markers Notum and Sall1 in the near cervical loop mesenchymal area, indicating that odontoblast fate selection happens before embedding into the odontoblastic layer (Figs.…”

“…Fos CreERT2 /R26 ZsGreen1 strain was used for genetic tracing of outer enamel epithelium progenitor cells. DMP1-Cherry/DSPP-cerulean mice were used for visualization of alveolar bone 38,49 . Acta2 CreERT2 /R26 tdTomato mice were used for lineage tracing of Acta2 + dental epithelial stem cells in cervical loop 31 .…”

Wnt/β-Catenin Signaling Promotes the Formation of Preodontoblasts In Vitro

“…All experimental protocols involving animal tissues in the present study were approved by the Institutional Animal Care and use Committee of UConn Health Center. DSPP-Cerulean/DMP1-Cherry, BSP-GFPtpz, pOBCol2.3FP (referred to as 2.3-GFP), TCF/Lef:H2B-EGFP, and nontransgenic mice used in this study have been previously described (Balic, Aguila, Caimano, et al 2010; Ferrer-Vaquer et al 2010; Vijaykumar et al 2019, 2020). All mice were maintained in the CD1 background.…”

“…pOBCol2.3-GFP (2.3-GFP) transgene is activated at early stages of odontoblast differentiation (preodontoblasts and prior to the expression of Dmp1 and Dspp ) (Balic, Aguila, and Mina 2010). DMP1-Cherry and DSPP-Cerulean transgenes are activated and expressed by functional and fully differentiated odontoblasts, respectively (Vijaykumar et al 2019). More recently, we showed that BSP-GFPtpz identifies cells in the osteogenic lineage and is excluded from cells in the dentinogenic lineage (Vijaykumar et al 2020).…”

Hsa-miRNA-143-3p Regulates Differentiation of Human Stem Cells From The Apical Papilla Through Targeting Nuclear Factor I-C

“…Odontoblasts synthesize and secrete dentin extracellular matrix proteins, among which DSPP and DMP1 are considered as typical marker for the odontogenic differentiation of mesenchyma stem cells [26] , and DMP1 might regulate the expression of DSPP [27] . Our results showed that the expression of NFIC, DSPP and KLF4 were inhibited and DMP1 was promoted by miR-143-3p.…”