Dataset Information

Limpet teeth LC-MSMS - The Microstructure, Proteomics and Crystallization of the Limpet Teeth

ABSTRACT: Limpets are marine mollusks using mineralized teeth, one of the hardest and strongest biomaterials, to feed on algae on intertidal rocks. However, most of studies only focuses on the ultrastructure and chemical composition of the teeth while the molecular information is largely unknown, limiting our understanding of this unique and fundamental biomineralization process. In this study, we investigated the teeth of limpet Cellana toreuma from three perspectives: 1) by using electron microscopy to observe the microstructure of the teeth; 2) by using proteomics and RNA-seq to investigate the proteins involved in the limpet teeth and 3) by in vitro crystallization experiment combined with Raman spectroscopy to investigate the effects of proteins and chitin framework on crystal formation. It is found that the limpets formed alternatively tricuspid teeth and unicuspid teeth. Small nanoneedles were densely packed at the tips or leading regions of the cusps. In contrast, big nanoneedles resembling chemical synthesized goethite were loosely packed in the trailing regions of the cusps. Proteins extracted from the whole teeth such as ferritin, peroxiredoxin, arginine kinase, GTPase-Rabs and clathrin were identified by proteomics. Goethite-binding experiment coupled with proteomics and RNA-seq highlighted six chitin-binding proteins (CtCBPs). Furthermore, these proteins or the framework chitin only induced packing of crystals without affecting their crystal polymorphs in vitro. Taken together, the limpets formed hierarchical teeth across different length scales through preformed framework and secreted complex proteins; in addition, the chitin could also be an important player in controlling crystallinity and crystal packing in vivo. This study provides insight into the unique biomineralization process in the limpet teeth at the molecular levels, which may guide biomimetic strategies aimed at designing hard materials at room temperature.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Cellana Toreuma

TISSUE(S): Radula, Somatic Cell

SUBMITTER: Yadong Wang

LAB HEAD: Rongqing Zhang

PROVIDER: PXD010113 | Pride | 2019-03-26

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

Items per page:

1 - 5 of 16

Publications

The Microstructure, Proteomics and Crystallization of the Limpet Teeth.

Wang Yadong Y Liu Chuang C Du Jinzhe J Huang Jingliang J Zhang Shuce S Zhang Rongqing R

Proteomics 20180905 19

Limpets are marine mollusks that use mineralized teeth, one of the hardest and strongest biomaterials, to feed on algae on intertidal rocks. However, most of studies only focus on the ultrastructure and chemical composition of the teeth while the molecular information is largely unknown, limiting our understanding of this unique and fundamental biomineralization process. The study investigates the microstructure, proteomics, and crystallization in the teeth of limpet Cellana toreuma. It is found ...[more]

PMID: 30142254

Similar Datasets

Project description:Deciduous and permanent human teeth represent a model system to study ageing of mesenchymal populations. Aging is tightly connected to self-renewal and proliferation and thus, mapping potential molecular differences in these characteristics between populations constitutes an important task. Specifically designed microarray panels were used. We have detected a number of molecules that were differentially expressed in dental pulp mesenchyme from deciduous and permanent teeth extracted from young children and adults, respectively. Among the differentially regulated genes HMGA2, a stem cell-associated marker, stood out as a remarkable example with a robust expression in deciduous pulp cells. In addition to this, we discovered that several proliferation-related genes, including CDC2A and CDK4, were up-regulated in deciduous pulp cells, while matrix genes COL1A1, fibronectin and several signaling molecules, such as VEGF, FGFr-1 and IGFr-1 were up-regulated in the pulp cells from permanent teeth. Taken together, our data suggest that deciduous pulp cells are more robust in self- renewal and proliferation, whereas adult dental pulp cells are more capable of signaling and matrix synthesis. The deciduous teeth were collected from 3-12-year-old children (N=8), whereas the permanent teeth were collected from adults, 19-52 years of age (N=8). The tooth was mechanically separated into two halves to remove all soft tissue in the pulp. The pulp tissue was digested and the dissociated cells were passed through cell strainer and plated with culture medium. The cells were then passaged at 90% confluency to a 10 cm dish for expansion. RNA was extracted from passage one cells with the Qiagen RNeasy Mini Kit. Microarray analysis was performed using PIQORTM Stem Cell Microarray chip (Miltenyi Biotech), which is comprised of 942 relevant genes for stem/progenitor cells and key genes for cell differentiation, to identify gene sets that are differentially expressed between cells in the deciduous and adult teeth. A total of 1 μg RNA for each sample was used for amplification and further analysis with the PIQORTM stem cell microarray chip (Miltenyi Biotec), followed by detection with a laser scanner (Agilent Technologies). In total, 8 comparisons were performed with dataset consisting of two microarrays, each containing a total of 11 slides with signal intensities within the R/Bioconductor statistical framework. The Linear Model for Microarray Data package (Limma) was used to pre-process the raw data, perform quality controls and estimate statistical significance. Expression intensities were background-corrected using a convolution of normal and exponential distributions with an offset of 50, which were then normalized within every slide using loess normalization. We analyzed the red and green channels separately using the mixed model method and different groups were quantile-normalized separately. The correlation was computed between the two channels for the same spot. Differentially expressed genes were identified by computing the contrasts, fitting to a linear model, performing the hypothesis tests, and correcting for multiple testing by performing the false discovery rate using the Benjamini-Hochberg correction.

Project description:Chitons are marine molluscs that posses a radula, a tongue like appendage containing many rows of microscopic teeth used for feeding. These teeth are hardened through the incorporation of biominerals. Since the original discovery of iron oxide and magnetite in these teeth, a wealth of chemical and structural knowledge has accumulated. The biomineralization process is known to be matrix mediated with precise control of the deposition of a number of different iron and calcium minerals at readily identifiable stages of tooth development. While much is known about the mechanisms of tooth mineralisation, there have been no studies undertaken to identify the genes that regulate this process. This investigation uses microarray technology to analyse the spatial expression of 493 expressed sequence tags (EST) derived from the radula sac of chiton, Acanthopleura hirtosa. A number of ESTs have been deemed significantly differentially expressed in relation to three designated areas of the radula sac. These spatial sections are defined by the variation in biomineral deposits of the radula, being immature radula which lacks any biomineral incorporation, iron deposition and calcium deposition. Additionally, foot muscle was used as a control to specify whether EST expression was specific to the radula and therefore likely involved biomineralization. A total of twelve A. hirtosa specimens were collected and sacrificed for microarray hybridizations. The radulae were removed from each animal immediately following sacrifice and dissected into three sections based on the biomineralisation development of the radula teeth. These sections are termed immature teeth, iron deposited teeth, and calcium deposited teeth. An additional sample of A. hirtosa foot muscle was taken from each specimen. RNA was extracted individually from these sections. Extracted RNA samples across subjects were pooled equally within sections in order to minimize the impact of biological variation and maximize the number of individual samples included in the study. To this effect, four single channel hybridizations were conducted for each radula and muscle section each representing a pooled biological replicate containing equal proportions of RNA from three A. hirtosa individuals. A total of sixteen microarray hybridizations were conducted in this manner.

			Action	DRS
	MS171763-1.raw	Raw
	MS171763-1.xlsx	Xlsx
	MS171763-2.raw	Raw
	MS171763-2.xlsx	Xlsx
	MS171763-3.raw	Raw