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Transcriptome analysis provides insights into the skeletal malformation induced by dietary phospholipids deficiency in largemouth bass (Micropterus salmoides) larvae

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Abstract

Largemouth bass is one of the most important freshwater aquaculture species in China. However, the mechanisms underlying the development of skeleton in the teleosts are not fully comprehensive. In a previous study in our laboratory, it was found that the addition of phospholipids to the diet significantly reduced the malformation rate of largemouth bass larvae. The malformation rate decreased from 91.29 ± 1.05% in the fish fed phospholipid-deficient diet (SP_L) to 11.99 ± 1.19% in the fish fed with diet including 8% phospholipids (SP_H). Therefore, high-throughput RNA-Seq was used to compare the transcriptome of largemouth bass larvae between SP_H and SP_L. Thirty whole body samples originated from 3 tanks in 8% phospholipids inclusion group and 3 tanks in no phospholipids inclusion group were used to reduce inaccuracies. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses revealed that the differentially expressed genes were involved in the mitogen-activated protein kinase (MAPK) signaling pathway, phosphatidylinositol signaling system, glycosphingolipid biosynthesis-lacto and neolacto series, and fatty acid degradation. Among the 584 differentially expressed genes, we paid attention to 6 genes known to be involved in skeletal development. Twist-related protein 2 (twist2) and dishevelled-associated activator of morphogenesis 1 (daam1), two genes related to osteoblast development, were up-regulated in SP_H. Bone gamma-carboxyglutamate (gla) protein (bglap), gene associated with the skeletal development and osteohormatology, was also up-regulated in SP_H. Procollagen C-endopeptidase enhancer b (pcolce b), a gene related to the development of gristle, was up-regulated in SP_H. Higher expression of secretory calcium-binding phosphoprotein 1 and 7 (scpp1 and scpp7) in SP_H suggests the stimulation of process related with tooth and bone development. The trend changes of the above genes all indicated that the lack of phospholipids might affect the skeletal development through the regulation of the expression of genes involved in several biological processes related to bone formation. In summary, those results provided valuable information about the reduction of deformity rates in largemouth bass and contributed to our understanding of the molecular mechanisms and regulative pathways regulating skeletal growth in larval carnivorous fish.

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Data availability

The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found below: NCBI SRA accession no. SRR25667165-25,667,170.

References

  • Anders S, Huber W (2012) Differential expression of RNA-Seq data at the gene level–the DESeq package. Heidelberg, Germany: EMBL 10:f1000research

  • Balla T, Gulyas G, Kim YJ, Pemberton J (2020) Phosphoinositides and calcium signaling; a marriage arranged at ER-PM contact sites. Curr Opin Physiol 17:149–157

    Article  PubMed  PubMed Central  Google Scholar 

  • Bennett CN, Ouyang H, Ma YL, Zeng Q, Gerin I, Sousa KM, Lane TF, Krishnan V, Hankenson KD, MacDougald OA (2007) Wnt10b increases postnatal bone formation by enhancing osteoblast differentiation. J Bone Miner Res 22:1924–1932

    Article  CAS  PubMed  Google Scholar 

  • Boglione C, Gagliardi F, Scardi M, Cataudella S (2001) Skeletal descriptors and quality assessment in larvae and post-larvae of wild-caught and hatchery-reared gilthead sea bream (Sparus aurata L. 1758). Aquaculture 192:1–22

    Article  Google Scholar 

  • Bolger AM, Lohse M, Usadel B (2014) Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics 30:2114–2120

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bonjour JP (2011) Calcium and phosphate: a duet of ions playing for bone health. J Am Coll Nutr 30:438S-448S

    Article  CAS  PubMed  Google Scholar 

  • Cahu C, Zambonino Infante J, Takeuchi T (2003a) Nutritional components affecting skeletal development in fish larvae. Aquaculture 227:245–258

    Article  CAS  Google Scholar 

  • Cahu CL, Infante JLZ, Barbosa V (2003b) Effect of dietary phospholipid level and phospholipid:neutral lipid value on the development of sea bass (Dicentrarchus labrax) larvae fed a compound diet. Brit J Nutr 90:21–28

    Article  CAS  PubMed  Google Scholar 

  • Conesa A, Götz S, García-Gómez JM, Terol J, Talón M, Robles M (2005) Blast2GO: a universal tool for annotation, visualization and analysis in functional genomics research. Bioinformatics 21:3674–3676

    Article  CAS  PubMed  Google Scholar 

  • Daprà F, Geurden I, Corraze G, Bazin D, Zambonino-Infante J, Fontagné-Dicharry S (2011) Physiological and molecular responses to dietary phospholipids vary between fry and early juvenile stages of rainbow trout (Oncorhynchus mykiss). Aquaculture 319:377–384

    Article  Google Scholar 

  • Du S, Haga Y (2004) The zebrafish as a model for studying skeletal development. Biomineralization: From Biology to Biotechnology and Medical Application, pp 283-304

  • Ferraresso S, Milan M, Pellizzari C, Vitulo N, Reinhardt R, Canario AV, Patarnello T, Bargelloni L (2010) Development of an oligo DNA microarray for the European sea bass and its application to expression profiling of jaw deformity. BMC Genomics 11:354

    Article  PubMed  PubMed Central  Google Scholar 

  • Fjelldal PG, Nordgarden U, Berg A, Grotmol S, Totland GK, Wargelius A, Hansen T (2005) Vertebrae of the trunk and tail display different growth rates in response to photoperiod in Atlantic salmon, Salmo salar L., post-smolts. Aquaculture 250:516–524

    Article  Google Scholar 

  • Fukumoto S, Iwamoto T, Sakai E, Yuasa K, Fukumoto E, Yamada A, Hasegawa T, Nonaka K, Kato Y (2006) Current topics in pharmacological research on bone metabolism: osteoclast differentiation regulated by glycosphingolipids. J Pharmacol Sci 100:195–200

    Article  CAS  PubMed  Google Scholar 

  • Geurden I, Marion D, Charlon N, Coutteau P, Bergot P (1998) Comparison of different soybean phospholipidic fractions as dietary supplements for common carp, Cyprinus carpio, larvae. Aquaculture 161:225–235

    Article  CAS  Google Scholar 

  • Ghosh-Choudhury N, Mandal CC, Choudhury GG (2007) Statin-induced ras activation integrates the phosphatidylinositol 3-kinase signal to akt and mapk for bone morphogenetic protein-2 expression in osteoblast differentiation. J Biol Chem 282:4983–4993

    Article  CAS  PubMed  Google Scholar 

  • Go W, Korzh V (2013) Plasma membrane Ca2+ ATPase Atp2b1a regulates bone mineralization in zebrafish. Bone 54:48–57

    Article  CAS  PubMed  Google Scholar 

  • Haga Y, Suzuki T, Takeuchi T (2002) Retinoic acid isomers produce malformations in postembryonic development of the Japanese flounder, Paralichthys olivaceus. Zool Sci 19:1105–1112

    Article  CAS  Google Scholar 

  • Haga Y, Du S, Satoh S, Kotani T, Fushimi H, Takeuchi T (2011) Analysis of the mechanism of skeletal deformity in fish larvae using a vitamin A-induced bone deformity model. Aquaculture 315:26–33

    Article  CAS  Google Scholar 

  • Hamza N, Mhetli M, Khemis IB, Cahu C, Kestemont P (2008) Effect of dietary phospholipid levels on performance, enzyme activities and fatty acid composition of pikeperch (Sander lucioperca) larvae. Aquaculture 275:274–282

    Article  CAS  Google Scholar 

  • Hattori M, Sawada Y, Kurata M, Yamamoto S, Kato K, Kumai H (2004) Oxygen deficiency during somitogenesis causes centrum defects in red sea bream, Pagrus major (Temminck et Schlegel). Aquac Res 35:850–858

    Article  Google Scholar 

  • Hoang QQ, Sicheri F, Howard AJ, Yang DS (2003) Bone recognition mechanism of porcine osteocalcin from crystal structure. Nature 425:977–980

    Article  CAS  PubMed  Google Scholar 

  • Huang Y, Xu J, Sheng Z, Chen N, Li S (2021) Integrated response of growth performance, fatty acid composition, antioxidant responses and lipid metabolism to dietary phospholipids in hybrid grouper (Epinephelus fuscoguttatus ♀ × E. lanceolatus ♂) larvae. Aquaculture 541:736728

    Article  CAS  Google Scholar 

  • Imsland AK, Foss A, Koedijk R, Folkvord A, Stefansson SO, Jonassen TM (2006) Short- and long-term differences in growth, feed conversion efficiency and deformities in juvenile Atlantic cod (Gadus morhua) startfed on rotifers or zooplankton. Aquac Res 37:1015–1027

    Article  Google Scholar 

  • Kanazawa A, Teshima S, Inamori S, Iwashita T, Naoao A (1981) Effects of phospholipids on growth, survival rate, and incidence of malformation in the larval ayu. Mem Fac Fish Kagoshima Univ 30:301–309

    CAS  Google Scholar 

  • Kawasaki K (2009) The SCPP gene repertoire in bony vertebrates and graded differences in mineralized tissues. Dev Genes Evol 219:147–157

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kolstad K, Thorland I, Refstie T, Gjerde B (2006) Genetic variation and genotype by location interaction in body weight, spinal deformity and sexual maturity in Atlantic cod (Gadus morhua) reared at different locations off Norway. Aquaculture 259:66–73

    Article  Google Scholar 

  • Kumar S, Connor JR, Dodds RA, Halsey W, Van Horn M, Mao J, Sathe G, Mui P, Agarwal P, Badger AM, Lee JC, Gowen M, Lark MW (2001) Identification and initial characterization of 5000 expressed sequenced tags (ESTs) each from adult human normal and osteoarthritic cartilage cDNA libraries. Osteoarthr Cartilage 9:641–653

    Article  CAS  Google Scholar 

  • Lall SP, Lewis-McCrea LM (2007) Role of nutrients in skeletal metabolism and pathology in fish — an overview. Aquaculture 267:3–19

    Article  CAS  Google Scholar 

  • Lerner UH, Ohlsson C (2015) The WNT system: background and its role in bone. J Intern Med 277:630–649

    Article  CAS  PubMed  Google Scholar 

  • Li S, Sang C, Wang A, Zhang J, Chen N (2019) The impacts of dietary carbohydrate levels on growth performance, feed utilization, glycogen accumulation and hepatic glucose metabolism in hybrid grouper (Epinephelus fuscoguttatus ♀ × E. lanceolatus ♂). Aquaculture 512:734351

    Article  CAS  Google Scholar 

  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using realtime quantitative PCR and the 2-ΔΔCT method. Methods 25:402–408

    Article  CAS  PubMed  Google Scholar 

  • Mazurais D, Darias MJ, Gouillou-Coustans MF, Le Gall MM, Huelvan C, Desbruyères E, Quazuguel P, Cahu C, Zambonino-Infante JL (2007) Dietary vitamin mix levels influence the ossification process in European sea bass (Dicentrarchus labrax) larvae. Am J Physiol Regul Integr Comp Physiol 294(2):R520-7

    Article  PubMed  Google Scholar 

  • Meunier FJ, Huysseune A (1992) The concept of bone tissue in Osteichthyes. Neth J Zool 42(2–3):445–458

    Google Scholar 

  • Ministry of Agriculture and Rural Affairs China (2020) China Fishery Statistics Yearbook. China Agriculture Press, Beijing

    Google Scholar 

  • Nguyen VT, Satoh S, Haga Y, Fushimi H, Kotani T (2008) Effect of zinc and manganese supplementation in Artemia on growth and vertebral deformity in red sea bream (Pagrus major) larvae. Aquaculture 285:184–192

    Article  CAS  Google Scholar 

  • Pertea M, Kim D, Pertea GM, Leek JT, Salzberg SL (2016) Transcript-level expression analysis of RNA-seq experiments with HISAT, StringTie and Ballgown. Nat Protoc 11:1650–1667

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Pertea M, Pertea GM, Antonescu CM, Chang T, Mendell JT, Salzberg SL (2015) StringTie enables improved reconstruction of a transcriptome from RNA-seq reads. Nat Biotechnol 33:290–295

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Pinto JP, Conceição N, Gavaia PJ, Cancela ML (2003) Matrix Gla protein gene expression and protein accumulation colocalize with cartilage distribution during development of the teleost fish Sparus aurata. Bone 32:201–210

    Article  CAS  PubMed  Google Scholar 

  • Pombinho ANR, Laiz V, Molha DM, Marques SMP, Cancela ML (2004) Development of two bone-derived cell lines from the marine teleost Sparus aurata; evidence for extracellular matrix mineralization and cell-type-specific expression of matrix Gla protein and osteocalcin. Cell Tissue Res 315:393–406

    Article  CAS  PubMed  Google Scholar 

  • Puvanendran V, Calder-Crewe C, Brown JA (2009) Vertebral deformity in cultured Atlantic cod larvae: ontogeny and effects on mortality. Aquac Res 40:1653–1660

    Article  Google Scholar 

  • Saavedra M, Barr Y, Pousão-Ferreira P, Helland S, Yúfera M, Dinis MT, Conceição LEC (2009) Supplementation of tryptophan and lysine in Diplodus sargus larval diet: effects on growth and skeletal deformities. Aquac Res 40:1191–1201

    Article  CAS  Google Scholar 

  • Salhotra A, Shah HN, Levi B, Longaker MT (2020) Mechanisms of bone development and repair. Nat Rev Mol Cell Biol 21:696–711

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sandel E, Nixon O, Lutzky S, Ginsbourg B, Tandler A, Uni Z, Koven W (2010) The effect of dietary phosphatidylcholine/phosphatidylinositol ratio on malformation in larvae and juvenile gilthead sea bream (Sparus aurata). Aquaculture 304:42–48

    Article  CAS  Google Scholar 

  • Seidel R, Jayasankar AK, Shahar R, Dean MN (2019) The multiscale architectures of fish bone and tessellated cartilage and their relation to function. Architect Mate Nat Eng Archimats:329–353

  • Silva TS, Cordeiro O, Richard N, Conceição LEC, Rodrigues PM (2011) Changes in the soluble bone proteome of reared white seabream (Diplodus sargus) with skeletal deformities. Comp Biochem Physiol D: Genom Proteom 6:82–91

    Google Scholar 

  • Simes DC, Williamson MK, Ortiz Delgado JB, Viegas C, Price PA, Cancela ML (2003) Purification of matrix gla protein from a marine teleost fish, argyrosomus regius: Calcified cartilage and not bone as the primary site of mgp Accumulation in Fish. J Bone Miner Res 18:244–259

    Article  CAS  PubMed  Google Scholar 

  • Stiassny ML (2000) Skeletal system. In: The laboratory fish. Academic Press, pp 109–118

  • Taylor JF, Martinez-Rubio L, Del Pozo J, Walton JM, Tinch AE, Migaud H, Tocher DR (2015) Influence of dietary phospholipid on early development and performance of Atlantic salmon (Salmo salar). Aquaculture 448:262–272

    Article  CAS  Google Scholar 

  • Thouverey C, Caverzasio J (2015) Focus on the p38 MAPK signaling pathway in bone development and maintenance. BoneKEy Reports 4:711

  • Tocher DR, Bendiksen EÅ, Campbell PJ, Bell JG (2008) The role of phospholipids in nutrition and metabolism of teleost fish. Aquaculture 280:21–34

    Article  CAS  Google Scholar 

  • Uyan O, Koshio S, Ishikawa M, Uyan S, Ren T, Yokoyama S, Komilus CF, Michael FR (2007) Effects of dietary phosphorus and phospholipid level on growth, and phosphorus deficiency signs in juvenile Japanese flounder, Paralichthys olivaceus. Aquaculture 267:44–54

    Article  CAS  Google Scholar 

  • Wang S, Han Z, Turchini GM, Wang X, Fang Z, Chen N, Xie R, Zhang H, Li S (2022) Effects of dietary phospholipids on growth performance, digestive enzymes activity and intestinal health of largemouth bass (Micropterus salmoides) larvae. Front Immunol 12:827946

  • Wauquier F, Léotoing L, Philippe C, Spilmont M, Coxam V, Wittrant Y (2015) Pros and cons of fatty acids in bone biology. Prog Lipid Res 58:121–145

    Article  CAS  PubMed  Google Scholar 

  • Whyte MP, Obrecht SE, Finnegan PM, Jones JL, Podgornik MN, McAlister WH, Mumm S (2002) Osteoprotegerin deficiency and juvenile Paget’s disease. N Engl J Med 347:175–184

    Article  CAS  PubMed  Google Scholar 

  • Yamaoka K, Inoue K, Tanuguchi T (1994) Territory behavior of hatchery-reared red sea bream juveniles and its relations to density and body deformity. Rep Inst Kuroshio Sphere Kochi Univ 8:21–25

    Google Scholar 

  • Yasutake J, Inohaya K, Kudo A (2004) Twist functions in vertebral column formation in medaka, Oryzias latipes. Mech Develop 121:883–894

    Article  CAS  Google Scholar 

  • Zhao J, Ai Q, Mai K, Zuo R, Luo Y (2013) Effects of dietary phospholipids on survival, growth, digestive enzymes and stress resistance of large yellow croaker, Larmichthys crocea larvae. Aquaculture 410–411:122–128

    Article  Google Scholar 

  • Zheng CX, Sui BD, Qiu XY, Hu CH, Jin Y (2020) Mitochondrial regulation of stem cells in bone homeostasis. Trends Mol Med 26:89–104

    Article  CAS  PubMed  Google Scholar 

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Funding

This work was financially supported by the National Key Research and Development Program of China (2022YFD2400803, 2019YFD0900203).

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Zishuo Fang; Investigation, Data curation, Formal analysis, Writing-Original Draft; Ye Gong; Investigation, Methodology; Shilin Wang; Investigation; Zhihao Han; Investigation, Methodology; Xuxiong Huang; Conceptualization; Funding acquisition; Naisong Chen; Conceptualization; Project administration; Songlin Li; Conceptualization, Supervision, Writing-Review & Editing, Funding acquisition.

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Correspondence to Songlin Li.

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The present experiment was strictly followed the requirement of Animal Care and Use Committee of Shanghai Ocean University (SHOU-DW-2022–026).

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Fang, Z., Gong, Y., Wang, S. et al. Transcriptome analysis provides insights into the skeletal malformation induced by dietary phospholipids deficiency in largemouth bass (Micropterus salmoides) larvae. Aquacult Int (2023). https://doi.org/10.1007/s10499-023-01359-2

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