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Molecular influences relating to the development and biomineralization of the chiton radula, Acanthopleura histosa

ABSTRACT: 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.

ORGANISM(S): Liolophura hirtosa

PROVIDER: GSE20105 | GEO | 2010/06/25

SECONDARY ACCESSION(S): PRJNA124235

REPOSITORIES: GEO

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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.

Project description:The dentition of elasmobranchs (sharks, skates and rays) is uniquely productive, capable of both rapid and continuous, lifelong regeneration. Elasmobranchs represent an important group of vertebrates with a deep evolutionary history, possessing several ancient and basal characters, i.e., the continuously regenerative dentition from a specialized dental lamina. The dental lamina is an expanded component of the oral epithelia that is responsible for initiating and producing new teeth among all toothed-vertebrates. In sharks, this dynamic epithelial unit is permanent and continuous – meaning it extends to cover the entirety of each jaw (jaw-wide) and develops early during embryogenesis and retained to produce teeth for the life of the shark. It is rare for a truly embryonic vertebrate tissue to be retained for its original function for the life of the organism. The dental lamina in sharks is unique and houses teeth in a developmental series from the deepest part, where teeth are initiated, through stages of tooth development in the form of a related, family of teeth to eruption and functionality of the advanced teeth at the jaw margin. How teeth are made and regenerated is an important question in vertebrate biology; here we investigated this question in the small spotted catshark (Scyliorhinus canicula), a new model in the field of developmental biology. Specifically, we divided the shark dental lamina into stage-compartments as follows: (i) the initiation site – the successional lamina (SL); (ii) the early developing teeth (ET); (iii) the late stage developing teeth (LT); (iv) the tooth-taste junction between the superficial oral and dental epithelium at the jaw margin that separates the taste territory and the dental lamina proper (TTJ); and basi-hyal oral epithelium that is strictly non-dental and only contains taste buds (BHTB). These 5 compartments each house both a shared and unique signature of gene transcripts. This study aims to understand the transcriptomic basis of continuous tooth regeneration in the shark. In this study we combine X-ray computed tomography, classic histology, insitu hybridization, immunohistochemistry, and functional assays of novel markers, and de novo and genome guided transcriptome assemblies for each of these 5 dental lamina compartments of the hatchling (stage 34) catshark (S. canicula).

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Molecular influences relating to the development and biomineralization of the chiton radula, Acanthopleura histosa

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