Quantitative proteomic analysis of deciduous molars in miniature pig (second repetition)
Ontology highlight
ABSTRACT:
INSTRUMENT(S): Q Exactive
ORGANISM(S): Sus Scrofa Domesticus (domestic Pig)
SUBMITTER: 
li yang
LAB HEAD: songlin wang
PROVIDER: PXD006174 | Pride | 2018-05-23
REPOSITORIES: Pride
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Project description:The miniature pig is diphyodont, making it a valuable alternative model for understanding human tooth development and replacement. However, little is known about gene expression and function during swine odontogenesis. The goal of this study is to undertake the survey of differential gene expression profiling with Affymetrix Porcine GeneChip and functional network analysis during morphogenesis of diphyodont dentition in miniature pigs. The identification of genes related to diphyodont development should lead to a better understanding of morphogenetic patterns and the mechanisms of diphyodont replacement in large animal models and humans. The staged miniature pig embryos and fetuses were obtained by cesarean section at E40, E50, and E60. The last deciduous molar germs, including the dental lamina in mandibles from the same litter were isolated and pooled. The gestational age of embryos used in our study just cover 3 characteristic stages: Dm3 in cap stage without secondary dental lamina (E40); Dm3 in bell stage with secondary dental lamina initiation (E50); Dm3 in secretory stage without evidence of morphological changes in the secondary dental lamina.
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Project description:Miniature pigs, a valuable alternative model for understanding human tooth development, have deciduous teeth from all four tooth families that are replaced once by permanent molars. The extracellular matrix (ECM) supports cells and maintains the integrity of tooth germs during tooth development. However, details on the role of the ECM in tooth development are poorly understood. Here, we performed long non- coding RNA (lncRNA) and messenger RNA (mRNA) expression profiles in the ECM components of deciduous tooth germs by RNA sequencing in miniature pigs. From the early-cap to the late-bell stages, we identified 4,562 and 3,238 differentially expressed genes (DEGs) from E40 to E50 and E50 to E60, respectively. In addition, a total of 1,464 differentially expressed lncRNAs from E40 to E50, and 969 differentially expressed lncRNAs from E50 to E60 were obtained. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis showed that DEGs were enriched significantly for multiple signaling pathways, especially for the ECM pathway. We then outlined the detailed dynamic gene expression profiling of ECM components during deciduous molar development. Comparison of the cap and bell stages revealed that the structure and functions of the ECM dynamically changed. The ECM-related genes, including THBS1, COL4A5, COL4A6, COL1A1, CHAD, TNR, GP1BA, and ITGA3, were significantly changed, and some were shown to enrich during the bell stage development. Finally, we outlined the co-expression of lncRNAs and ECM properties during tooth development. We showed that the interplay of key lncRNAs could change ECM processes and influence the ECM establishment of tooth patterns to accomplish full tooth formation. These results might provide information to elucidate the regulation network of the lncRNA and ECM in tooth development.
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Project description:Gingival Crevicular Fluid, a plasma-derived exudate present in the gingival crevice was collected from deciduous, exfoliating and permanent teeth from 20 children (60 samples) with the aim to characterize and quantify by a mass spectrometry based top-down proteomic approach, the peptide/proteins in the fluid and verify possible variations occurring during the exfoliating process. The results obtained confirmed the presence in Gingival Crevicular Fluid of α-Defensins 1-4, Thymosin β4 and Thymosin β10, as described in previous works and revealed the presence of other interesting peptides never described before in Gingival Crevicular Fluid, such as specific fragments of α-1-antitrypsin, α-1-antichymotrypsin, Thymosin β4 and Thymosin β10 fragments, Fibrinopeptide A, Fibrinopeptide B, S100A, LVV Hemorphin-7 (hemoglobin chain β fragment), as well as some other peptides deriving from α and β subunits of hemoglobin. Statistical analysis evidenced different levels in 5 proteins/peptides in the three groups in particular with higher level in exfoliating teeth. Our study demonstrate that an in-depth analysis of a biological fluid like Gingival Crevicular Fluid, present in small amount, can provide useful information for the understanding of different biological processes like teeth eruption.
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Project description:Human deciduous and permanent teeth exhibit different developmental processes, morphologies, histological characteristics and life cycles. In addition their pulp tissues react differently to external stimuli, such as the pulp sensitivity test, dental trauma and pulp therapy materials. These differences are attributable to their genetic backgrounds. Therefore the purpose of this study is to compare the differences of dental pulp in deciduous and permanent teeth. Pulp samples were obtained from permanent premolars (n=6, aged 11-14 years) and deciduous teeth (n=6, aged 11-14 years). Comparative cDNA microarrary analysis revealed several differences in gene expression between the deciduous and permanent pulp tissues. Each GSM record represents a pulp sample pooled from two teeth samples.
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Project description:The miRNAs expression profile of three different types of teeth include deciduous incisor (QY), deciduous canine (JY) , deciduous premolar (QMY) ,and deciduous molar (MY) in three typical stages of tooth development embryonic day 40 , 50, and 60, which cover the major morphological and physiological changes in pig tooth germ growth and development throughout pregnancy including the bud, cap, and bell stages. twelve-condition experiment, QY40 vs.QY50 vs.QY60 vs. JY40 vs. JY50vs. JY60 vs.QMY40 vs.QMY50 vs.QMY60 vs.MY40.vs.MY50.vs.MY60. Biological replicates: 1 , independently removed under a microscope. Four replicate per array.
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Project description:There are histological and functional differences between human deciduous and permanent pediodontal ligament (PDL) tissues. The purpose of this study was to determine the differences between these two types of tissue at the molecular level by comparative gene expression analysis. PDL samples were obtained from permanent premolars (n=38) and anterior deciduous teeth (n=31) extracted from 40 healthy persons. Comparative cDNA microarrary analysis revealed several differences in gene expression between the deciduous and permanent PDL tissues.
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Project description:We identified the spatiotemporal pattern of cascade initiation of additional molars in miniature pig,where second molar (M2) initiated from the posterior-free end of the dental lamina over the first molar (M1) at E60 when M1 progressed to the late bell stage. Similarly, third molar (M3) budded off from the posterior-free end of the dental lamina over M2, which reached bell stage at PN20. However, the molecular mechanisms of the regulatory network during sequential formation of additional molars remain poorly characterized in diphyodont mammals. We performed microarrays on miniature pigs at three molar developmental stages to examine their differential gene expression profiles and potential regulatory networks during additional molar morphogenesis.
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