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Effects of Dietary Cystine and Tyrosine Supplementation on Melanin Synthesis in the Pacific Oyster (Crassostrea gigas)

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Abstract

Melanogenesis is a multistep process to produce melanin for dark pigmentation in skin coloration. Previous studies in vertebrates demonstrated that cystine and tyrosine amino acids are involved in the melanin synthesis. However, very little is known about the melanogenesis in bivalve. In this study, cystine supplementation for 30 days significantly upregulated the expression of CgB-aat1, CgCbs and CgTyr and pheomelanin content in the Pacific oyster Crassostrea gigas. Transmission electron microscope (TEM) results revealed more melanosomes in the connective tissue and melanin granules were secreted in epithelium of mantle. In contrast, tyrosine supplementation had no clear effect on melanogenesis except the gene expression changes of CgB-aat1 and CgCbs. In addition, prolonged supplementation of cystine or tyrosine for 60 days had a negative impact on melanogenesis. Indeed, after 60 days, expression of most of the melanin synthesis-related genes under study was decreased, and melanin content was significantly reduced, indicating that cystine and tyrosine might inhibit production of eumelanin and pheomelanin, respectively. In addition, in vitro analysis using primary cell culture from mantle tissue indicated that incubation with cystine, tyrosine, or B-AAT1 polypeptide, CBS/TYR recombinant proteins induced the increase of CgB-aat1 and CgCbs expression in a dose-dependent manner, suggesting the presence of a regulatory network in response to cystine and tyrosine amino acids intakes in pheomelanin synthesis-related gene expression. Taken together, these data indicate that cystine-CgB-aat1-CgCbs-CgTyr axis is a potential regulator of the pheomelanin biosynthesis pathway, and thus plays an important role in the mantle pigmentation in C. gigas. This work provides a new clue for selective cultivation of oyster strains with specific shell colors in bivalve breeding.

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Data Availability

The datasets generated in the current study are available from the corresponding author on reasonable request.

Abbreviations

B-aat1 :

B(0, +)-type amino acid transporter 1

Cbs :

Cystathionine beta-synthase

Ef1α :

α Subunit of elongation factor 1

PMSF:

Phenylmethanesulfonyl fluoride

RIPA:

Radio immunoprecipitation assay

Tyr :

Tyrosinase

Tyrp1 :

Tyrosinase-related protein 1

Tyrp2 :

Tyrosinase-related protein 2

WB:

Western blot

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Acknowledgements

This work was supported by the grants from the China Agriculture Research System Project (CARS-49), National Natural Science Foundation of China (31972789), and the Earmarked Fund for Agriculture Seed Improvement Project of Shandong Province (2021LZGC027, 2020LZGC016).

Funding

This work was supported by the grants from the China Agriculture Research System Project (CARS-49), National Natural Science Foundation of China (31972789), and the Earmarked Fund for Agriculture Seed Improvement Project of Shandong Province (2021LZGC027, 2020LZGC016).

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Authors and Affiliations

  1. Key Laboratory of Mariculture, Ministry of Education, Ocean University of China, Qingdao, 266003, China

    Zhuanzhuan Li, Chengxun Xu, Hong Yu, Lingfeng Kong, Shikai Liu & Qi Li

  2. Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao, 266237, China

    Qi Li

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  1. Zhuanzhuan Li
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  2. Chengxun Xu
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  3. Hong Yu
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  4. Lingfeng Kong
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Contributions

Qi Li: Experimental design and coordination, manuscript revision and funding acquisition. Zhuanzhuan Li: Completion of the experiment, data analysis and manuscript drafting. Chengxun Xu: Resources. Hong Yu: Investigation. Lingfeng Kong: Data curation. Shikai Liu: Supervision.

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Correspondence to Qi Li.

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The pacific oyster is neither an endangered nor protected species. All experiments in this study were conducted according to national and institutional guidelines.

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Li, Z., Xu, C., Yu, H. et al. Effects of Dietary Cystine and Tyrosine Supplementation on Melanin Synthesis in the Pacific Oyster (Crassostrea gigas). Mar Biotechnol 25, 537–547 (2023). https://doi.org/10.1007/s10126-023-10223-6

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