, 2011) is to produce xeno-grafted pearl sacs from two closely related species where inter-specific sequence differences in homologous biomineralisation genes are present. Mantle grafts between two species, P. maxima and P. margaritifera (so called xenografts), have previously been shown to result in pearl sac formation and pearl development ( McGinty et al., 2010). Where species-specific gene differences are present between
these species for homologous biomineralisation genes, then the use of xenografts can be used to unequivocally ascertain whether the host or donor cells are transcriptionally selleck chemicals active for the relevant gene through detecting the species-specific transcript present. The expression of donor oyster putative biomineralisation genes (N = 2) within the pearl sac at the time of pearl collection has previously been
verified (McGinty et al., 2011). In the present study, species diagnostic single nucleotide polymorphisms (SNPs) were developed using high-throughput mRNA sequencing (Illumina, GAII) derived from allografted P. maxima and P. margaritifera pearl sacs to detect putative biomineralisation genes expressed in pearl sac tissue. Based on the use of this improved technology and the analysis of more genes from previous work, the present study aims to determine whether host or donor derived cells are primarily responsible for the expression of biomineralisation INCB024360 in vivo genes in pearl sac tissue. Adult P. margaritifera and P. maxima pearl oysters were sourced from wild populations in West Papuan Province (1°13′N, 130°54′E), and from a hatchery in Bali (8°23′S, 115°14E), Indonesia,
respectively. Both Carnitine palmitoyltransferase II oyster species are native to the Indo-Pacific region ( Gervis and Sims, 1992). Three months prior to nucleus implantation, P. margaritifera oysters were shipped to Bali in a commercial pearl oyster transport boat, placed into 16 pocket-panel nets suspended on longlines and allowed to adjust to environmental conditions at the Bali site. Net panels were covered with mesh to reduce oyster metabolic rate and gametogenic activity three weeks prior to seeding to reduce the chance of implanted nuclei rejection ( Gervis and Sims, 1992). Allografted and xenografted oysters were produced as reported in McGinty et al. (2010). Briefly, 80 P. maxima and 80 P. margaritifera host oysters were implanted with either allograft (Ss, Bb) or xenograft (Sb, Bs) mantle tissues ( Fig. 1). Ten P. maxima and 10 P. margaritifera oysters were used as mantle tissue donors. Excised mantle tissue from each oyster was cut into eight pieces, with four pieces used as allografts (species controls) and four as xenografts (experimental treatments). Appropriately sized seed nuclei were chosen according to the gonad size of the host oyster (ranging from 5.76–7.88 mm and 0.28–0.