Comparative proteomics of mantle from Mytilous coruscus
Ontology highlight
ABSTRACT:
ORGANISM(S): Mytilus Coruscus
SUBMITTER: 
Zhi Liao
PROVIDER: PXD034858 | iProX | Thu Jun 23 00:00:00 BST 2022
REPOSITORIES: iProX
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Project description:The mantle is a thin tissue from which proteins are secreted dictating the mollusk shell construction. As a conserved organ involved in shell formation throughout mollusks, the mantle is an excellent foundation from which to study biomineralization. A P. maxima mantle tissue specific cDNA microarray, termed PmaxArray 1.0, has been developed comprising 5000 cDNA transcripts derived from the mantle tissue of P. maxima. This tool has been used to investigate the spatial functional dynamics of the mantle tissue identifying over 2000 PmaxArray 1.0 spots as differentially expressed spatially within this organ. Gene expression profiles observed for these transcripts indicated 5 major spatial functions for the mantle, 3 of which have been putatively attributed to shell formation roles associated with nacre microstructure, calcite prismatic microstructure and periostracum. These transcripts are further examined with in situ expression localization and comparative sequence analyses in reference to potential shell formation roles. This spatial investigation has expedited the elucidation of functions within the dynamic mantle organ, paying particular attention to of shell biomineralization. Keywords: Spatial expression profiling by array
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Project description:The mantle is a thin tissue from which proteins are secreted dictating the mollusk shell construction. As a conserved organ involved in shell formation throughout mollusks, the mantle is an excellent foundation from which to study biomineralization. A P. maxima mantle tissue specific cDNA microarray, termed PmaxArray 1.0, has been developed comprising 5000 cDNA transcripts derived from the mantle tissue of P. maxima. This tool has been used to investigate the spatial functional dynamics of the mantle tissue identifying over 2000 PmaxArray 1.0 spots as differentially expressed spatially within this organ. Gene expression profiles observed for these transcripts indicated 5 major spatial functions for the mantle, 3 of which have been putatively attributed to shell formation roles associated with nacre microstructure, calcite prismatic microstructure and periostracum. These transcripts are further examined with in situ expression localization and comparative sequence analyses in reference to potential shell formation roles. This spatial investigation has expedited the elucidation of functions within the dynamic mantle organ, paying particular attention to of shell biomineralization. Keywords: Spatial expression profiling by array The mantle tissue from 9 animals was dissected into 5 separate sections: outer fold (OF), middle fold (MF), inner fold (IF), ventral mantle tissue (VM) and dorsal mantle tissue (DM). Total RNA was extracted from these tissues and pooled across subjects in order to reduce the effect of biological variation; such that 3 individuals were pooled together totaling 3 pooled replicate samples for each tissue. All the biologically pooled tissue types were compared against a common reference in which total RNA from all tissues types and all nine animals was equally pooled. A total of 30 dual channel microarrays hybridizations were performed and analyzed.
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Project description:Specimens of L. elliptica were collected by scuba divers at a depth of 10-18m in January 2010 at Hangar Cove, Rothera Point, Adelaide Island, Antarctic Peninsula (67°34’07°S, 68°07’30°W). Animals were collected in two size classes: large animals (with lengths ranging around 60mm) and small animals (lengths ranging around 30mm), the sizes of which equated to average ages of 16 and 7 years respectively. These two groups were termed “old” and “young” respectively. The clams were maintained in a flow-through aquarium and allowed to acclimate to laboratory conditions for 2 weeks before experimentation. At the end of the acclimation period, 10 old and 10 young animals were transferred to a 60l jacketed tank with aerated sea water, connected to a thermocirculator. The sea water temperature was gradually raised from 0°C to +3°C over a 12 hour period. This temperature was then maintained for a further 12 hours, before sampling the animals. Mantle tissue samples were dissected and immediately snap frozen in liquid nitrogen and stored at -80°C. The sampling regime was repeated on 10 old and 10 young animals that had been maintained in the flow-through aquarium for the same time period (control animals) RNA was extracted from all samples using TriSure, according to manufacturer’s instructions. RNAs from mantle tissue from 5 animals from each group (old treated, young treated, old control and young control) were used in the array hybridization experiments. The array used was A-MEXP-1676. PCR amplified labelled cDNA targets were prepared from 1μg total RNA. 5 replicates were used with 1 dye swap.
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