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CDNA microarray analysis revealing candidate biomineralizational genes of the pearl oyster, Pinctada fucata martensii

ABSTRACT: In order to screen and identify biomineralization gene, microarray technique was used to reveal tissues specific expression genes in the brunet mantle edge (ME), mantle centre (MC), and both ME and MC (ME-MC) from assembled transcriptome contigs of Pinctada fucata martensii, ideal pearl oyster for the study of biomineralization. Tissues of ME, MC, hepatopancreas, foot, gill, adductor muscle, heart and intestine were sampled from two females and one male pearl oyster. Gonad was sampled from above three individuals and another two male and one female. Equal amount RNA of each individual hepatopancreas, foot, gill, adductor muscle, heart and intestine were mixed as a composite viscera sample (CV), and equal amount gonad RNA from one male and one female were combined together as a gonad sample (GS). Hybridizations were performed with twelve samples of ME, MC, CV and GS.

ORGANISM(S): Pinctada imbricata

PROVIDER: GSE70439 | GEO | 2016/07/01

SECONDARY ACCESSION(S): PRJNA288694

REPOSITORIES: GEO

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Project description:Research using the oyster Crassostrea gigas as a model has experienced a rapid growth in recent years thanks to the development of high throughput molecular technologies. As many as 56,268 EST sequences have so far been sequenced, representing a genome-wide resource that can be used for microarray investigations. We have developed an oyster microarray containing cDNAs representing 31,918 unique transcribed sequences. The genes spotted on the array have been selected from the publicly accessible GigasDatabase established from cDNA libraries derived from a wide variety of tissues and different developmental stages. n this paper, we report the transcriptome of male and female gonads, mantle, gills, posterior adductor muscle, visceral ganglion, hemocytes, labial palps and digestive gland. Following validation of the microarray, statistical analyses were used to identify genes differentially expressed among tissues and define clusters of tissue-specific genes. These genes reflect well major tissue-specific functions at the molecular level. Analysis of hierarchical clustering data also predicted the involvement of un-annotated genes in selected functional pathways. In a second instance, microarray data were used to accurately select housekeeping genes common to all tissues. Their expression profiles was compared to common oyster standard genes used for quantitative RT-PCR calibration (actin, g3apdh and ef1α). The novel candidate housekeeping gene, adp-ribosylation factor 1 (arf1) and g3apdh gene seem to be more robust for normalizing gene expression data of tissues. This study provides a new source for annotating the oyster genome. It also identified new candidate housekeeping genes, a prerequisite for accurate quantitative RT-PCR expression profiling. Gene expression was measured in 9 tissues: Female gonad, male gonad, mantle, gills, posterior adductor muscle, visceral ganglion, hemocytes, labial palps, digestive gland.Three to four biological replicates were analysed per tissue. These were from distinct animals for female gonad, male gonad, mantle, gills, posterior adductor muscle, labial palps and digestive glands, or obtained from a pool of 6 individuals for hemocytes and visceral ganglion.

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CDNA microarray analysis revealing candidate biomineralizational genes of the pearl oyster, Pinctada fucata martensii

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