|
|
线纹海马Dmrt1基因的克隆与表达分析
|
Abstract:
Dmrt1广泛参与了不同物种的性别决定和性腺发育过程，为探究dmrt1在线纹海马中的分子特征及表达模式，本研究采用RACE-PCR克隆到了其cDNA全长1009 bp，包含87 bp的5’UTR、756 bp的ORF和166 bp的3’UTR，共编码251个氨基酸；氨基酸多重比对显示，线纹海马dmrt1在DM结构域内具有高度保守性；系统进化树分析表明，其与侏儒海马的dmrt1亲缘关系最近；RT-qPCR显示，dmrt1在线纹海马成体的性腺中呈显著的雄性偏好性高表达于精巢，表明其可能参与了线纹海马精巢的形成和维持。该研究为进一步深入研究线纹海马dmrt1在性别发育中的功能奠定了基础。
Dmrt1 is widely involved in the process of sex determination and gonad development in different species. In order to investigate the molecular characteristics and expression pattern of dmrt1 in the Hippocampus erectus, an 1009 bp cDNA was cloned by RACE-PCR, which contains 756 bp ORF, 87 bp 5’UTR, 166 bp 3’ UTR, encoding 251 amino acids. A highly conserved DM domain has been detected in the dmrt1 of Hippocampus erectus by multiple amino acid alignment. Phylogenetic tree analysis showed dmrt1 in Hippocampus erectus was evolutionarily most closely related to that in Hippocampus zosterae. A male-biased expression in testis was detected by RT-qPCR, suggesting that dmrt1 might play important roles in the formation and maintenance of testis in Hippocampus erectus. This study laid the foundation to further study the function of dmrt1 in sexual development of Hippocampus erectus.
| [1] | Raymond, C.S., Shamu, C.E., Shen, M.M., Seifert, K.J., Hirsch, B., Hodgkin, J., et al. (1998) Evidence for Evolutionary Conservation of Sex-Determining Genes. Nature, 391, 691-695. https://doi.org/10.1038/35618 |
| [2] | Erdman, S.E. and Burtis, K.C. (1993) The Drosophila Doublesex Proteins Share a Novel Zinc Finger Related DNA Binding Domain. The EMBO Journal, 12, 527-535. https://doi.org/10.1002/j.1460-2075.1993.tb05684.x |
| [3] | Hodgkin, J. (2002) The Remarkable Ubiquity of DM Domain Factors as Regulators of Sexual Phenotype: Ancestry or Aptitude? Genes & Development, 16, 2322-2326. https://doi.org/10.1101/gad.1025502 |
| [4] | Kopp, A. (2012) Dmrt Genes in the Development and Evolution of Sexual Dimorphism. Trends in Genetics, 28, 175-184. https://doi.org/10.1016/j.tig.2012.02.002 |
| [5] | Matson, C.K. and Zarkower, D. (2012) Sex and the Singular DM Domain: Insights into Sexual Regulation, Evolution and Plasticity. Nature Reviews Genetics, 13, 163-174. https://doi.org/10.1038/nrg3161 |
| [6] | 孙业盈, 张全启, 齐洁, 王志刚, 陈妍婕, 李春梅, 钟其旺. 半滑舌鳎DMRT1基因的克隆与表达分析[J]. 武汉大学学报(理学版), 2008(2): 221-226. |
| [7] | Cui, Z., Liu, Y., Wang, W., Wang, Q., Zhang, N., Lin, F., et al. (2017) Genome Editing Reveals Dmrt1 as an Essential Male Sex-Determining Gene in Chinese Tongue Sole (Cynoglossus semilaevis). Scientific Reports, 7, Article No. 42213. https://doi.org/10.1038/srep42213 |
| [8] | Kobayashi, T., Kajiura-Kobayashi, H., Guan, G. and Nagahama, Y. (2007) Sexual Dimorphic Expression of DMRT1 and Sox9a during Gonadal Differentiation and Hormone-Induced Sex Reversal in the Teleost Fish Nile Tilapia (Oreochromis niloticus). Developmental Dynamics, 237, 297-306. https://doi.org/10.1002/dvdy.21409 |
| [9] | Otake, H., Shinomiya, A., Matsuda, M., Hamaguchi, S. and Sakaizumi, M. (2006) Wild-Derived XY Sex-Reversal Mutants in the Medaka, Oryzias latipes. Genetics, 173, 2083-2090. https://doi.org/10.1534/genetics.106.058941 |
| [10] | Sato, T., Endo, T., Yamahira, K., Hamaguchi, S. and Sakaizumi, M. (2005) Induction of Female-to-Male Sex Reversal by High Temperature Treatment in Medaka, Oryzias latipes. Zoological Science, 22, 985-988. https://doi.org/10.2108/zsj.22.985 |
| [11] | Yamamoto, T. (1958) Artificial Induction of Functional Sex-Reversal in Genotypic Females of the Medaka (Oryzias latipes). Journal of Experimental Zoology, 137, 227-263. https://doi.org/10.1002/jez.1401370203 |
| [12] | Guo, Y., Cheng, H., Huang, X., Gao, S., Yu, H. and Zhou, R. (2005) Gene Structure, Multiple Alternative Splicing, and Expression in Gonads of Zebrafish Dmrt1. Biochemical and Biophysical Research Communications, 330, 950-957. https://doi.org/10.1016/j.bbrc.2005.03.066 |
| [13] | Webster, K.A., Schach, U., Ordaz, A., Steinfeld, J.S., Draper, B.W. and Siegfried, K.R. (2017) Dmrt1 Is Necessary for Male Sexual Development in Zebrafish. Developmental Biology, 422, 33-46. https://doi.org/10.1016/j.ydbio.2016.12.008 |
| [14] | Xu, S., Zhang, S., Zhang, W., Liu, H., Wang, M., Zhong, L., et al. (2022) Genome-Wide Identification, Phylogeny, and Expression Profile of the Dmrt (Double Sex and Mab-3 Related Transcription Factor) Gene Family in Channel Catfish (Ictalurus punctatus). Frontiers in Genetics, 13, Article ID: 891204. https://doi.org/10.3389/fgene.2022.891204 |
| [15] | 闫宁宁. 大口黑鲈Dmrt1基因的克隆与表达分析[D]: [硕士学位论文]. 上海: 上海海洋大学, 2019. |
| [16] | Johnsen, H., Seppola, M., Torgersen, J.S., Delghandi, M. and Andersen, ？. (2010) Sexually Dimorphic Expression of Dmrt1 in Immature and Mature Atlantic Cod (Gadus morhua L.). Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology, 156, 197-205. https://doi.org/10.1016/j.cbpb.2010.03.009 |
| [17] | Fritzsche, R.A. and Vincent, A. (2002) The Living Marine Resources of the Western Central Atlantic. Food and Agriculture Organization of the United Nations, Rome, 1221-1225. |
| [18] | Foster, S.J. and Vincent, A.C.J. (2004) Life History and Ecology of Seahorses: Implications for Conservation and Management. Journal of Fish Biology, 65, 1-61. https://doi.org/10.1111/j.0022-1112.2004.00429.x |
| [19] | 陈灵珍, 秦耿, 王信, 刘雅莉, 肖旺红, 林强. 线纹海马(Hippocampus erectus)性腺特异性结构与生殖细胞发育特征研究[J]. 热带海洋学报, 2020, 39(6): 93-102. |
| [20] | Matson, C.K., Murphy, M.W., Sarver, A.L., Griswold, M.D., Bardwell, V.J. and Zarkower, D. (2011) DMRT1 Prevents Female Reprogramming in the Postnatal Mammalian Testis. Nature, 476, 101-104. https://doi.org/10.1038/nature10239 |
| [21] | Raymond, C.S., Murphy, M.W., O’Sullivan, M.G., Bardwell, V.J. and Zarkower, D. (2000) Dmrt1, a Gene Related to Worm and Fly Sexual Regulators, Is Required for Mammalian Testis Differentiation. Genes & Development, 14, 2587-2595. https://doi.org/10.1101/gad.834100 |
| [22] | Raymond, C.S., Kettlewell, J.R., Hirsch, B., Bardwell, V.J. and Zarkower, D. (1999) Expression of Dmrt1 in the Genital Ridge of Mouse and Chicken Embryos Suggests a Role in Vertebrate Sexual Development. Developmental Biology, 215, 208-220. https://doi.org/10.1006/dbio.1999.9461 |
| [23] | Lei, N., Hornbaker, K.I., Rice, D.A., Karpova, T., Agbor, V.A. and Heckert, L.L. (2007) Sex-Specific Differences in Mouse DMRT1 Expression Are Both Cell Type-and Stage-Dependent during Gonad Development. Biology of Reproduction, 77, 466-475. https://doi.org/10.1095/biolreprod.106.058784 |
| [24] | Kim, S., Bardwell, V.J. and Zarkower, D. (2007) Cell Type-Autonomous and Non-Autonomous Requirements for Dmrt1 in Postnatal Testis Differentiation. Developmental Biology, 307, 314-327. https://doi.org/10.1016/j.ydbio.2007.04.046 |
| [25] | Omotehara, T., Smith, C.A., Mantani, Y., Kobayashi, Y., Tatsumi, A., Nagahara, D., et al. (2014) Spatiotemporal Expression Patterns of Doublesex and Mab-3 Related Transcription Factor 1 in the Chicken Developing Gonads and Müllerian Ducts. Poultry Science, 93, 953-958. https://doi.org/10.3382/ps.2013-03672 |
| [26] | Cooper, C.A., Challagulla, A., Jenkins, K.A., Wise, T.G., O’Neil, T.E., Morris, K.R., et al. (2016) Generation of Gene Edited Birds in One Generation Using Sperm Transfection Assisted Gene Editing (Stage). Transgenic Research, 26, 331-347. https://doi.org/10.1007/s11248-016-0003-0 |
| [27] | Smith, C.A., Roeszler, K.N., Ohnesorg, T., Cummins, D.M., Farlie, P.G., Doran, T.J., et al. (2009) The Avian Z-Linked Gene DMRT1 Is Required for Male Sex Determination in the Chicken. Nature, 461, 267-271. https://doi.org/10.1038/nature08298 |
| [28] | Osawa, N., Oshima, Y. and Nakamura, M. (2005) Molecular Cloning of Dmrt1 and Its Expression in the Gonad of Xenopus. Zoological Science, 22, 681-687. https://doi.org/10.2108/zsj.22.681 |
| [29] | Matsuda, M., Nagahama, Y., Shinomiya, A., Sato, T., Matsuda, C., Kobayashi, T., et al. (2002) DMY Is a Y-Specific Dm-Domain Gene Required for Male Development in the Medaka Fish. Nature, 417, 559-563. https://doi.org/10.1038/nature751 |
| [30] | 王静杰. 胡子鲶Dmrt1基因全长cDNA的克隆和时空表达[D]: [硕士学位论文]. 湛江: 广东海洋大学, 2011. |
| [31] | 李林, 梁宏伟, 李忠, 罗相忠, 张志伟, 朱媛媛, 邹桂伟. 黄颡鱼DMRT1基因cDNA全长克隆及其表达分析[J]. 华中农业大学学报, 2012, 31(2): 220-226. |
