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Molecular characterization and expression analysis of Patched 1 gene in the half-smooth tongue sole (Cynoglossus semilaevis)

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Abstract

The signaling molecule hedgehog (Hh) is essential for cellular signaling required for body pattern formation in embryonic and individual development in vertebrate and invertebrates. Patched 1 (Ptc1), a receptor of Hh, mediates the activity of Hh by binging the Hh ligands. In this study, the Ptc1 of half-smooth tongue sole Cynoglossus semilaevis (CsPtc1) was cloned and characterized. The full-length of CsPtc1 cDNA is 5 212 nucleotides (nt), and encodes a protein with 1 543 amino acid residues. CsPtc1 shared many features with Ptc1 of other telesot species. Real-time quantitative PCR revealed CsPtc1 mRNA expressed in brain, liver, heart, gill, intestines, spleen, gonad and kidney, with expression level in testis significantly higher than in ovary. In testis, hybridization signals were mainly detected in primary spermatocytes, secondary spermatocytes, and sertoli cells, while weak signals were found in oocytes. Correspondingly, the degree of methylation is higher in female than in male and pseudomale. Results indicate that CsPtc1 may be involved in Desert Hedgehog (DHH) maintenance of the male and pseudo-male germ line and spermatogenesis.

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References

  • Alcedo J, Ayzenzon M, Von Ohlen T, et al. 1996. The Drosophila smoothened gene encodes a seven-pass membrane protein, a putative receptor for the hedgehog signal. Cell, 86(2): 221–232

    Article  Google Scholar 

  • Barsoum I B, Bingham N C, Parker K L, et al. 2009. Activation of the Hedgehog pathway in the mouse fetal ovary leads to ectopic appearance of fetal Leydig cells and female pseudohermaphroditism. Developmental Biology, 329(1): 96–103

    Article  Google Scholar 

  • Beaudoing E, Freier S, Wyatt J R, et al. 2000. Patterns of variant polyadenylation signal usage in human genes. Genome Research, 10(7): 1001–1010

    Article  Google Scholar 

  • Bellvé A R, Millette C F, Bhatnagar Y M, et al. 1977. Dissociation of the mouse testis and characterization of isolated spermatogenic cells. Journal of Histochemistry & Cytochemistry, 25(7): 480–494

    Article  Google Scholar 

  • Bitgood M J, McMahon A P. 1995. Hedgehog and Bmp genes are coexpressed at many diverse sites of cell-cell interaction in the mouse embryo. Developmental Biology, 172(1): 126–138

    Article  Google Scholar 

  • Bitgood M J, Shen Liya, McMahon A P. 1996. Sertoli cell signaling by Desert hedgehog regulates the male germline. Current Biology, 6(3): 298–304

    Article  Google Scholar 

  • Canto P, Söderlund D, Reyes E, et al. 2004. Mutations in the desert hedgehog (DHH) gene in patients with 46, XY complete pure gonadal dysgenesis. The Journal of Clinical Endocrinology & Metabolism, 89(9): 4480–4483

    Article  Google Scholar 

  • Canto P, Vilchis F, Söderlund D, et al. 2005. A heterozygous mutation in the desert hedgehog gene in patients with mixed gonadal dysgenesis. Molecular Human Reproduction, 11(11): 833–836

    Article  Google Scholar 

  • Carpenter D, Stone D M, Brush J, et al. 1998. Characterization of two patched receptors for the vertebrate hedgehog protein family. Proceedings of the National Academy of Sciences of the United States of America, 95(23): 13630–13634

    Article  Google Scholar 

  • Carstea E D, Morris J A, Coleman K G, et al. 1997. Niemann-Pick C1 disease gene: homology to mediators of cholesterol homeostasis. Science, 277(5323): 228–231

    Article  Google Scholar 

  • Chen Songlin, Ji Xiangshan, Shao Changwei, et al. 2012. Induction of mitogynogenetic diploids and identification of WW super-female using sex-specific SSR markers in half-smooth tongue sole (Cynoglossus semilaevis). Marine Biotechnology, 14(1): 120–128

    Article  Google Scholar 

  • Chen Yongbin, Jiang Jin. 2013. Decoding the phosphorylation code in Hedgehog signal transduction. Cell Research, 23(2): 186–200

    Article  Google Scholar 

  • Chen Songlin, Li Jing, Deng Siping, et al. 2007. Isolation of femalespecific AFLP markers and molecular identification of genetic sex in half-smooth tongue sole (Cynoglossus semilaevis). Marine Biotechnology, 9(2): 273–280

    Article  Google Scholar 

  • Chen Yu, Struhl G. 1996. Dual roles for patched in sequestering and transducing Hedgehog. Cell, 87(3): 553–563

    Article  Google Scholar 

  • Chen Songlin, Zhang Guojie, Shao Changwei, et al. 2014. Whole-genome sequence of a flatfish provides insights into ZW sex chromosome evolution and adaptation to a benthic lifestyle. Nature Genetics, 46(3): 253–260

    Article  Google Scholar 

  • Clark A M, Garland K K, Russell L D. 2000. Desert hedgehog (Dhh) gene is required in the mouse testis for formation of adult-type Leydig cells and normal development of peritubular cells and seminiferous tubules. Biology of Reproduction, 63(6): 1825–1838

    Article  Google Scholar 

  • Concordet J P, Lewis K E, Moore J W, et al. 1996. Spatial regulation of a zebrafish patched homologue reflects the roles of sonic hedgehog and protein kinase A in neural tube and somite patterning. Development, 122(9): 2835–2846

    Google Scholar 

  • Daugela L, Nüsgen N, Walier M, et al. 2012. Measurements of DNA methylation at seven loci in various tissues of CD1 mice. PLoS One, 7(9): e44585

    Article  Google Scholar 

  • De Sousa P A, Caveney A, Westhusin M, et al. 1998a. Temporal patterns of embryonic gene expression and their dependence on oogenetic factors. Theriogenology, 49(1): 115–128

    Article  Google Scholar 

  • De Sousa P A, Watson A J, Schultz G A, et al. 1998b. Oogenetic and zygotic gene expression directing early bovine embryogenesis: a review. Molecular Reproduction and Development, 51(1): 112–121

    Article  Google Scholar 

  • Deng Siping, Chen Songion, Liu Benwei, et al. 2008. Molecular Cloning and Expression Analysis of FTZ-F1 in the Half-smooth Tongue-sole, Cynoglossus semilaevis. Zoological Research (in Chinese), 29(6): 592–598

    Article  Google Scholar 

  • Deng Siping, Chen Songlin, Tian Yongsheng, et al. 2007. Gonadal differentiation and effects of temperature on sex determination in half-smooth tongue-sole, Cynoglossus semilaevis. Journal of Fishery Sciences of China (in Chinese), 14(5): 714–719

    Google Scholar 

  • Deng Siping, Chen Songlin, Xu Jianyong, et al. 2009. Molecular cloning, characterization and expression analysis of gonadal P450 aromatase in the half-smooth tongue-sole, Cynoglossus semilaevis. Aquaculture, 287(1–2): 211–218

    Article  Google Scholar 

  • Dong Xiaoli, Chen Songlin, Ji Xiangshan, et al. 2011. Molecular cloning, characterization and expression analysis of Sox9a and Foxl2 genes in half-smooth tongue sole (Cynoglossussemilaevis). Acta Oceanologica Sinica, 30(1): 68–77

    Article  Google Scholar 

  • Hammerschmidt M, Brook A, McMahon A P. 1997. The world according to hedgebog. Trends in Genetics, 13(1): 14–21

    Article  Google Scholar 

  • Hu Qiaomu, Chen Songlin. 2013. Cloning, genomic structure and expression analysis of ubc9 in the course of development in the half-smooth tongue sole (Cynoglossus semilaevis). Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology, 165(3): 181–188

    Article  Google Scholar 

  • Huang C C J, Yao H H C. 2010. Diverse functions of Hedgehog signaling in formation and physiology of steroidogenic organs. Molecular Reproduction and Development, 77(6): 489–496

    Article  Google Scholar 

  • Huangfu D, Anderson K V. 2006. Signaling from Smo to Ci/Gli: conservation and divergence of Hedgehog pathways from Drosophila to vertebrates. Development, 133(1): 3–14

    Article  Google Scholar 

  • Ingham P W, McMahon A P. 2001. Hedgehog signaling in animal development: paradigms and principles. Genes & Development, 15(23): 3059–3087

    Article  Google Scholar 

  • Jacob L, Lum L. 2007. Hedgehog signaling pathway in Drosophila. Science’s STKE, 2007(407): cm7

    Google Scholar 

  • Jiang Yanwei, Wan Ruijing, Chen Ruisheng, et al. 1993. Studies on technique of artificial fry rearing of Cynoglossus semilaevis Günther in Bohai Sea. Marine Fisheries Research (in Chinese), (14): 25–33

    Google Scholar 

  • Johnson M, Zaretskaya I, Raytselis Y, et al. 2008. NCBI BLAST: a better web interface. Nucleic Acids Research, 36(S2): W5–W9

    Article  Google Scholar 

  • Kamisago M, Kimura M, Furutani Y, et al. 1999. Assignment1 of human desert hedgehog gene (DHH) to chromosome band 12q13 1 by in situ hybridization. Cytogenetic and Genome Research, 87(1–2): 117–118

    Article  Google Scholar 

  • Koebernick K, Hollemann T, Pieler T. 2001. Molecular cloning and expression analysis of the Hedgehog receptors XPtc1 and XSmo in Xenopus laevis. Mechanisms of Development, 100(2): 303–308

    Article  Google Scholar 

  • Kumar S, Balczarek K A, Lai Zhichun. 1996. Evolution of the hedgehog gene family. Genetics, 142(3): 965–972

    Google Scholar 

  • Lewis K, Concordet J P, Ingham P W. 1999. Characterisation of a Second patched Gene in the Zebrafish Danio rerio and the Differential Response of patched Genes to Hedgehog Signalling. Developmental Biology, 208(1): 14–29

    Article  Google Scholar 

  • 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

    Article  Google Scholar 

  • Loftus S K, Morris J A, Carstea E D, et al. 1997. Murine model of Niemann-Pick C disease: mutation in a cholesterol homeostasis gene. Science, 277(5323): 232–235

    Article  Google Scholar 

  • Ma Xuekun, Liu Xuezhou, Wen Haishen, et al. 2006. Histological observation on gonadal sex differentiation in Cynoglossus semilaevis Günther. Marine Fisheries Research (in Chinese), 27(2): 55–61

    Google Scholar 

  • Marigo V, Davey R A, Zuo Yi, et al. 1996. Biochemical evidence that patched is the Hedgehog receptor. Nature, 384(6605): 176–179

    Article  Google Scholar 

  • Nüsslein-Volhard C, Wieschaus E. 1980. Mutations affecting segment number and polarity in Drosophila. Nature, 287(5785): 795–801

    Article  Google Scholar 

  • O'Hara W A, Azar W J, Behringer R R, et al. 2011. Desert hedgehog is a mammal-specific gene expressed during testicular and ovarian development in a marsupial. BMC Developmental Biology, 11(1): 72

    Article  Google Scholar 

  • Park S Y, Meeks J J, Raverot G, et al. 2005. Nuclear receptors Sf1 and Dax1 function cooperatively to mediate somatic cell differentiation during testis development. Development, 132(10): 2415–2423

    Article  Google Scholar 

  • Petersen T N, Brunak S, von Heijne G, et al. 2011. SignalP 4.0: discriminating signal peptides from transmembrane regions. Nature Methods, 8(10): 785–786

    Article  Google Scholar 

  • Rahnama F, Toftgard R, Zaphiropoulos P G. 2004. Distinct roles of PTCH2 splice variants in Hedgehog signalling. Biochemical Journal, 378(2): 325–334

    Article  Google Scholar 

  • Russell M C, Cowan R G, Harman R M, et al. 2007. The hedgehog signaling pathway in the mouse ovary. Biology of Reproduction, 77(2): 226–236

    Article  Google Scholar 

  • Shao Changwei, Chen Songlin, Scheuring C F, et al. 2010. Construction of two BAC libraries from half-smooth tongue sole Cynoglossus semilaevis and identification of clones containing candidate sex-determination genes. Marine Biotechnology, 12(5): 558–568

    Article  Google Scholar 

  • Shao Changwei, Li Qiye, Chen Songlin, et al. 2014. Epigenetic modification and inheritance in sexual reversal of fish. Genome Research, 24(4): 604–615

    Article  Google Scholar 

  • Shao Jianguo, Zhang Ling, Gao Jun, et al. 2006. Aberrant expression of PTCH (patched gene) and Smo (smoothened gene) in human pancreatic cancerous tissues and its association with hyperglycemia. Pancreas, 33(1): 38–44

    Article  Google Scholar 

  • Sievers F, Higgins D G. 2014. Clustal Omega, accurate alignment of very large numbers of sequences. Multiple Sequence Alignment Methods, 1079: 105–116

    Article  Google Scholar 

  • Smyth I, Narang M A, Evans T, et al. 1999. Isolation and characterization of human patched 2 (PTCH2), a putative tumour suppressor gene in basal cell carcinoma and medulloblastoma on chromosome 1p32. Human Molecular Genetics, 8(2): 291–297

    Article  Google Scholar 

  • Song Wentao, Li Yangzhen, Zhao Yongwei, et al. 2012. Construction of a high-density microsatellite genetic linkage map and mapping of sexual and growth-related traits in half-smooth tongue sole (Cynoglossus semilaevis). PLoS One, 7(12): e52097

    Article  Google Scholar 

  • Stone D M, Hynes M, Armanini M, et al. 1996. The tumour-suppressor gene patched encodes a candidate receptor for Sonic hedgehog. Nature, 384(6605): 129–134

    Article  Google Scholar 

  • Suzuki M M, Bird A. 2008. DNA methylation landscapes: provocative insights from epigenomics. Nature Reviews Genetics, 9(6): 465–476

    Article  Google Scholar 

  • Szczepny A, Hime G R, Loveland K L. 2006. Expression of hedgehog signalling components in adult mouse testis. Developmental Dynamics, 235(11): 3063–3070

    Article  Google Scholar 

  • Takabatake T, Takahashi T C, Takabatake Y, et al. 2000. Distinct expression of two types of Xenopus Patched genes during early embryogenesis and hindlimb development. Mechanisms of Development, 98(1–2): 99–104

    Article  Google Scholar 

  • Tamura K, Peterson D, Peterson N, et al. 2011. MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Molecular Biology and Evolution, 28(10): 2731–2739

    Article  Google Scholar 

  • Umehara F, Tate G, Itoh K, et al. 2000. A Novel mutation of desert hedgehog in a patient with 46, XY partial gonadal dysgenesis accompanied by minifascicular neuropathy. The American Journal of Human Genetics, 67(5): 1302–1305

    Article  Google Scholar 

  • Varjosalo M, Taipale J. 2008. Hedgehog: functions and mechanisms. Genes & Development, 22(18): 2454–2472

    Article  Google Scholar 

  • Watson A J, Watson P H, Arcellana-Panlilio M, et al. 1994. A growth factor phenotype map for ovine preimplantation development. Biology of Reproduction, 50(4): 725–733

    Article  Google Scholar 

  • Wijgerde M, Ooms M, Hoogerbrugge J W. et al. 2005. Hedgehog signaling in mouse ovary: indian hedgehog and desert hedgehog from granulosa cells induce target gene expression in developing theca cells. Endocrinology, 146(8): 3558–3566

    Article  Google Scholar 

  • Wong K Y, Huang Xiaojun, Chim C S. 2012. DNA methylation of microRNA genes in multiple myeloma. Carcinogenesis, 33(9): 1629–1638

    Article  Google Scholar 

  • Yao H H C, Whoriskey W, Capel B. 2002. Desert Hedgehog/Patched 1 signaling specifies fetal Leydig cell fate in testis organogenesis. Genes & Development, 16(11): 1433–1440

    Article  Google Scholar 

  • Zhang Liyan, Liu Wanjun, Shao Changwei, et al. 2014. Cloning, expression and methylation analysis of piwil2 in half-smooth tongue sole (Cynoglossus semilaevis). Marine Genomics, 18: 45–54

    Article  Google Scholar 

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Correspondence to Songlin Chen.

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Foundation item: The National Natural Science Foundation under contract Nos 31130057 and 31472269; the Taishan Scholar Project Fund of Shandong of China.

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Dong, Z., Qi, Q., Zhang, N. et al. Molecular characterization and expression analysis of Patched 1 gene in the half-smooth tongue sole (Cynoglossus semilaevis). Acta Oceanol. Sin. 35, 19–28 (2016). https://doi.org/10.1007/s13131-016-0865-2

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  • DOI: https://doi.org/10.1007/s13131-016-0865-2

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