Cloning, expression profiling and promoter functional analysis of bone morphogenetic protein 2 in the tongue sole (Cynoglossus semilaevis)

MA Qian; FAN Yanjun; ZHUANG Zhimeng; LIU Shufang

doi:10.1007/s13131-018-1164-x

Article Contents

Article Navigation > Acta Oceanologica Sinica > 2018 > 37(2): 76-84

MA Qian, FAN Yanjun, ZHUANG Zhimeng, LIU Shufang. Cloning, expression profiling and promoter functional analysis of bone morphogenetic protein 2 in the tongue sole (Cynoglossus semilaevis)[J]. Acta Oceanologica Sinica, 2018, 37(2): 76-84. doi: 10.1007/s13131-018-1164-x

Citation:

MA Qian, FAN Yanjun, ZHUANG Zhimeng, LIU Shufang. Cloning, expression profiling and promoter functional analysis of bone morphogenetic protein 2 in the tongue sole (Cynoglossus semilaevis)[J]. Acta Oceanologica Sinica, 2018, 37(2): 76-84. doi: 10.1007/s13131-018-1164-x

Citation:

MA Qian, FAN Yanjun, ZHUANG Zhimeng, LIU Shufang. Cloning, expression profiling and promoter functional analysis of bone morphogenetic protein 2 in the tongue sole (Cynoglossus semilaevis)[J]. Acta Oceanologica Sinica, 2018, 37(2): 76-84. doi: 10.1007/s13131-018-1164-x

PDF( 1959 KB)

Cloning, expression profiling and promoter functional analysis of bone morphogenetic protein 2 in the tongue sole (Cynoglossus semilaevis)

doi: 10.1007/s13131-018-1164-x

1.
Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China;Function Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266200, China
2.
Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China
3.
Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China;Function Laboratory for Marine Biology and Biotechnolgy, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266200, China

Received Date: 2017-03-03
Rev Recd Date: 2011-11-07

Abstract

Abstract

BMP2 plays crucial roles in vertebrate developmental process and acts as a bone inducer during osteogenesis. We present here the molecular cloning of bmp2 cDNA from the marine flatfish Cynoglossus semilaevis, and the analysis of bmp2 expression profiling and promoter function. The full length of bmp2 cDNA sequence is 2 048 bp, which encodes a protein of 422 amino acids. Tissue expression distribution of bmp2 was examined in 14 tissues of mature individuals by quantitative real time PCR (qRT-PCR). The results revealed that bmp2 was expressed ubiquitously, and the highest expression level was detected in the spinal cord. Moreover, bmp2 expression levels were detected at 15 sampling time points of early developmental stages (egg, larva, juvenile and fingerling stages). The highest expression level of bmp2 was observed at the gastrula stage, which was about ten times higher than those at the other three embryo stages. Whole-mount in situ hybridization showed that the bmp2 signal was strongly detected at the location of the crown-like larval fin, heart and liver, and slightly expressed in the notochord at one day post hatch (dph); then the expression of bmp2 started to be concentrated in notochord at three dph. Subsequently, we characterized the 5'-flanking region of bmp2 by testing the promoter activity by Luciferase reporter assays. Positive regulatory region was detected at the location of -179 to +109. The predicted transcription factor binding sites (E-box binding factors, zinc finger transcription factor, etc.) in this region might participate in the transcriptional regulation of the bmp2 gene.
- cloning,
- gene expression pattern,
- promoter transcriptional activity,
- bone morphogenetic protein,
- Cynoglossus semilaevis,
- early developmental stages

FullText(HTML)

References(48)

References

Alexander C, Zuniga E, Blitz I L, et al. 2011. Combinatorial roles for BMPs and Endothelin 1 in patterning the dorsal-ventral axis of the craniofacial skeleton. Development, 138(23): 5135-5146

Balemans W, Van Hul W. 2002. Extracellular regulation of BMP signaling in vertebrates: a cocktail of modulators. Developmental Biology, 250(2): 231-250

Bragdon B, Moseychuk O, Saldanha S, et al. 2011. Bone morphogenetic proteins: a critical review. Cellular Signalling, 23(4): 609-620

Burkus J K, Heim S E, Gornet M F, et al. 2003. Is INFUSE bone graft superior to autograft bone? An integrated analysis of clinical trials using the LT-CAGE lumbar tapered fusion device. Journal of Spinal Disorders & Techniques, 16(2): 113-122

Chalazonitis A, Kessler J A. 2012. Pleiotropic effects of the bone morphogenetic proteins on development of the enteric nervous system. Developmental Neurobiology, 72(6): 843-856

Chandler R L, Chandler K J, McFarland K A, et al. 2007. Bmp2 transcription in osteoblast progenitors is regulated by a distant 3' enhancer located 156.3 kilobases from the promoter. Molecular and Cellular Biology, 27(8): 2934-2951

Cheng Hongwei, Jiang Wei, Phillips F M, et al. 2003. Osteogenic activity of the fourteen types of human bone morphogenetic proteins (BMPs). The Journal of Bone and Joint Surgery (American Volume), 85-A(8): 1544-1552

Dathe K, Kjaer K W, Brehm A, et al. 2009. Duplications involving a conserved regulatory element downstream of BMP2 are associated with brachydactyly type A2. The American Journal of Human Genetics, 84(4): 483-492

de Castro E, Sigrist C J A, Gattiker A, et al. 2006. ScanProsite: detection of PROSITE signature matches and ProRule-associated functional and structural residues in proteins. Nucleic Acids Research, 34(S2): W362-W365

Ducy P, Karsenty G. 2000. The family of bone morphogenetic proteins. Kidney International, 57(6): 2207-2214

Fernández I, Darias M, Andree K B, et al. 2011. Coordinated gene expression during gilthead sea bream skeletogenesis and its disruption by nutritional hypervitaminosis A. BMC Developmental Biology, 11: 7

Ghosh-Choudhury N, Choudhury G G, Harris M A, et al. 2001. Autoregulation of mouse BMP-2 gene transcription is directed by the proximal promoter element. Biochemical and Biophysical Research Communications, 286(1): 101-108

Graff J M. 1997. Embryonic patterning: to BMP or not to BMP, that is the question. Cell, 89(2): 171-174

Hammond K L, Loynes H E, Mowbray C, et al. 2009. A late role for bmp2b in the morphogenesis of semicircular canal ducts in the zebrafish inner ear. PLoS One, 4(2): e4368

Heng S, Paule S, Hardman B, et al. 2010. Posttranslational activation of bone morphogenetic protein 2 is mediated by proprotein convertase 6 during decidualization for pregnancy establishment. Endocrinology, 151(8): 3909-3917

Hochheimer A, Tjian R. 2003. Diversified transcription initiation complexes expand promoter selectivity and tissue-specific gene expression. Genes & Development, 17(11): 1309-1320

Hogan B L. 1996. Bone morphogenetic proteins: multifunctional regulators of vertebrate development. Genes & Development, 10(13): 1580-1594

Holland L Z, Holland P W, Holland N D, et al. 1996. Revealing homologies between body parts of distantly related animals by in situ hybridization to developmental genes: amphioxus versus vertebrates. In: Ferraris J, Palumbi S R, eds. Molecular Zoology: Advances, Strategies, and Protocols. New York: Wiley, 473-483

Huang Weibiao, Rudkin G H, Carlsen B, et al. 2002. Overexpression of BMP-2 modulates morphology, growth, and gene expression in osteoblastic cells. Experimental Cell Research, 274(2): 226-234

Kang Q, Sun M H, Cheng H, et al. 2004. Characterization of the distinct orthotopic bone-forming activity of 14 BMPs using recombinant adenovirus-mediated gene delivery. Gene Therapy, 11(17): 1312-1320

Kawai S, Sugiura T. 2001. Characterization of human bone morphogenetic protein (BMP)-4 and -7 gene promoters: activation of BMP promoters by Gli, a sonic hedgehog mediator. Bone, 29(1): 54-61

Kishimoto Y, Lee K H, Zon L, et al. 1997. The molecular nature of zebrafish swirl: BMP2 function is essential during early dorsoventral patterning. Development, 124(22): 4457-4466

Kugimiya F, Kawaguchi H, Kamekura S, et al. 2005. Involvement of endogenous bone morphogenetic protein (BMP) 2 and BMP6 in bone formation. Journal of Biological Chemistry, 280(42): 35704-35712

Kültz D. 2012. The combinatorial nature of osmosensing in fishes. Physiology, 27(4): 259-275

Ling Fei, Wang Tao, Wei Liqiong, et al. 2009. Cloning and characterization of the 5'-flanking region of the pig adiponectin gene. Biochemical and Biophysical Research Communications, 381(2): 236-240

Livak K J, Schmittgen T D. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2^-ΔΔCT Method. Methods, 25(4): 402-408

Ma Qian, Feng Wenrong, Zhuang Zhimeng, et al. 2017. Cloning, expression profiling and promoter functional analysis of Bone morphogenetic protein 6 and 7 in tongue sole (Cynoglossus semilaevis). Fish Physiology and Biochemistry, 43(2): 435-454,, doi: 10.1007/s10695-016-0298-z

Ma Qian, Zhuang Zhimeng, Feng Wenrong, et al. 2015. Evaluation of reference genes for quantitative real-time PCR analysis of gene expression during early development processes of the tongue sole (Cynoglossus semilaevis). Acta Oceanologica Sinica, 34(10): 90-97

Marques C L, Fernández I, Rosa J, et al. 2014. Spatiotemporal expression and retinoic acid regulation of bone morphogenetic proteins 2, 4 and 16 in Senegalese sole. Journal of Applied Ichthyology, 30(4): 713-720

Marques C L, Fernández I, Viegas M N, et al. 2016. Comparative analysis of zebrafish bone morphogenetic proteins 2, 4 and 16: molecular and evolutionary perspectives. Cellular and Molecular Life Sciences, 73(4): 841-857

Martínez-Barberá J P, Toresson H, Da Rocha S, et al. 1997. Cloning and expression of three members of the zebrafish Bmp family: Bmp2a, Bmp2b and Bmp4. Gene, 198(1-2): 53-59

Messeguer X, Escudero R, Farré D, et al. 2002. PROMO: detection of known transcription regulatory elements using species-tailored searches. Bioinformatics, 18(2): 333-334

Quint E, Smith A, Avaron F, et al. 2002. Bone patterning is altered in the regenerating zebrafish caudal fin after ectopic expression of sonic hedgehog and bmp2b or exposure to cyclopamine. Proceedings of the National Academy of Sciences of the United States of America, 99(13): 8713-8718

Rafael M S, Laizé V, Cancela M L. 2006. Identification of Sparus aurata bone morphogenetic protein 2: molecular cloning, gene expression and in silico analysis of protein conserved features in vertebrates. Bone, 39(6): 1373-1381

Sandelin A, Carninci P, Lenhard B, et al. 2007. Mammalian RNA polymerase Ⅱ core promoters: insights from genome-wide studies. Nature Reviews Genetics, 8(6): 424-436

Sato T, Mikawa S, Sato K. 2010. BMP2 expression in the adult rat brain. The Journal of Comparative Neurology, 518(22): 4513-4530

Sato Y, Nishida M. 2010. Teleost fish with specific genome duplication as unique models of vertebrate evolution. Environmental Biology of Fishes, 88(2): 169-188

Schlange T, Andrée B, Arnold H H, et al. 2000. BMP2 is required for early heart development during a distinct time period. Mechanisms of Development, 91(1-2): 259-270

Simon M, Feliers D, Arar M, et al. 2002. Cloning of the 5'-flanking region of the murine bone morphogenetic protein-7 gene. Molecular and Cellular Biochemistry, 233(1-2): 31-37

Smith A, Avaron F, Guay D, et al. 2006. Inhibition of BMP signaling during zebrafish fin regeneration disrupts fin growth and scleroblast differentiation and function. Developmental Biology, 299(2): 438-454

Sugiura T. 1999. Cloning and functional characterization of the 5'-flanking region of the human bone morphogenetic protein-2 gene. Biochemical Journal, 338(2): 433-440

Talbot W S, Hopkins N. 2000. Zebrafish mutations and functional analysis of the vertebrate genome. Genes & Development, 14(7): 755-762

Taylor J S, Van de Peer Y, Braasch I, et al. 2001. Comparative genomics provides evidence for an ancient genome duplication event in fish. Philosophical Transactions of the Royal Society B: Biological Sciences, 356(1414): 1661-1679

Urist M R, Mikulski A, Lietze A. 1979. Solubilized and insolubilized bone morphogenetic protein. Proceedings of the National Academy of Science of the United States of America, 76(4): 1828-1832

Wan Ruijing, Jiang Yanwei, Zhuang Zhimeng. 2004. Morphological and developmental characters at the early stages of the tonguefish Cynoglossus semilaevis. Acta Zoologica Sinica (in Chinese), 50(1): 91-102

Wang E A, Rosen V, D'Alessandro J S, et al. 1990. Recombinant human bone morphogenetic protein induces bone formation. Proceedings of the National Academy of Science of the United States of America, 87(6): 2220-2224

Xue Yu, Zheng Xiudeng, Huang Lei, et al. 2014. Organizer-derived Bmp2 is required for the formation of a correct Bmp activity gradient during embryonic development. Nature Communications, 5: 3766

Ying Ying, Zhao Guangquan. 2001. Cooperation of endoderm-derived BMP2 and extraembryonic ectoderm-derived BMP4 in primordial germ cell generation in the mouse. Developmental Biology, 232(2): 484-492

Relative Articles

Supplements(0)

Cited By

Proportional views