Comparison of gene expression profile and development of red spotted grouper (Epinephelus coioides) larvae fed with various highly unsaturated fatty acids enriched rotifer
Date Issued
2009
Date
2009
Author(s)
Wang, Chun-Ping
Abstract
In the early stage of grouper larvae culture, Rotifer (Brachionus pliicatilis) is the major living feed. However, the nutrition of rotifer is not stable, which leads to the lack of n-3 highly unsaturated fatty acid (HUFA) in larvae and the increase of malformation rate and mortality. In this study, we investigated the effect of rotifer nutrition enrichment on nutrition composition and skeleton development of Epinephelus coioides from the nutrition genomics point of view. We investigated the differentially expressed genes by microarray biochip to establish the gene markers relationship between healthy phenotype and genotype as the basis for nutrition needs and embryo development. There are two rotifer enrichment formulas designed in this study, one is Super DC Selco, the other is cod emulsion oil. After rotifer enrichment, the crude lipid content increases 0.79 % (Super DC Selco), 1.72 % (cod emulsion oil) in rotifer, and 0.48 % (Super DC Selco), 0.39 % (cod emulsion oil) in larvae. When the larvae group fed with rotifer enriched with emulsion, respectively the arachidonic acid (ARA) composition decreased. However, the accumulation amount of eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA) and n-3 highly unsaturated fatty acid (HUFA) were significantly increased. The n-3 HUFA of larvae fed rotifer increased 8.7 % enriched with Super DC Selco, and 8.9 % enriched with cod emulsion oil, respectively. The fatty acid transfer through the food chain between rotifer and larvae was proportional. The DHA/EPA ratio was above 1, no matter the larvae was fed with enriched- or un-enriched- rotifer, and the DHA/EPA ratio even reached 4 for larvae fed with rotifer enriched with Super DC Selco. Peroxisome proliferator-activated receptor γ (PPAR γ) is an important transcription factor in regulating gene expression of lipid metabolism. Compare the expression amount of PPAR γ from grouper larvae in enriched group and un-enriched group by RT-PCR, we found that there is no significant difference between each group, which indicated that the nutrition of un-enriched- or enriched- rotifer has high nutrition already. Using the microarray biochip made from grouper embryonic stage cDNA library to analyze the gene expression profile of larvae fed with rotifer enriched with Super DC Selco or cod emulsion oil or un-enriched rotifer, we found that there were 37 genes with 1.5 fold up-regulation and 27 genes with 1.5 fold down-regulation. Further NCBI blast results showed that the up-regulated genes include skeletal and muscle development related structural proteins：ex myosin light chain 3, actin alpha skeletal muscle and muscle creatine kinase; metabolism related enzymes：ex chymotrypsinogen 2, pyruvate kinase, pancreatic carboxypeptidase B1, enolase 3, apolipoprotein A and hypothetical protein LOC. Whereas down-regulated genes include energy metabolism related protein： cytochrome c oxidase subunit III；structural protein：alpha 2 type I collagen；DNA replication related protein：checkpoint kinase 1 and activin receptor IIA. It is concluded that grouper larvae fed with enriched- rotifer the total n-3 HUFA was increased compare to that fed with un-enriched- rotifer in the composition of crude lipid, EPA, DHA and n-3 HUFA. Therefore microarray technique can establish the nutrition composition of living feed correlation with development of grouper larvae.
Subjects
Grouper
Rotifer
n-3 HUFA
ARA
EPA
DHA
PPAR γ
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