Volume 35 Issue 4
Jul.  2014
Turn off MathJax
Article Contents

Hong-Mei ZHANG, Yan-Fang SU, Zhi-Yi SHI, Yuan-Shuai FU. cDNA clone and expression analysis of α-Tropomyosin during Japanese flounder (Paralichthys olivaceus) metamorphosis. Zoological Research, 2014, 35(4): 307-312. doi: 10.13918/j.issn.2095-8137.2014.4.307
Citation: Hong-Mei ZHANG, Yan-Fang SU, Zhi-Yi SHI, Yuan-Shuai FU. cDNA clone and expression analysis of α-Tropomyosin during Japanese flounder (Paralichthys olivaceus) metamorphosis. Zoological Research, 2014, 35(4): 307-312. doi: 10.13918/j.issn.2095-8137.2014.4.307

cDNA clone and expression analysis of α-Tropomyosin during Japanese flounder (Paralichthys olivaceus) metamorphosis

doi: 10.13918/j.issn.2095-8137.2014.4.307
  • Received Date: 2014-04-01
  • Rev Recd Date: 2014-04-01
  • Publish Date: 2014-07-08
  • Tropomyosin (TM) plays a critical role in skeletal and cardiac muscle development and function. To assess the functional significance of α-TM in Japanese flounder (Paralichthys olivaceus) development and metamorphosis, cDNA from Japanese flounder was cloned and α-TM mRNA measured during development and metamorphosis. The full-length cDNA is 1 191 bp, including a 5'-untranslated region of 114 bp, a 3'-UTR of 222 bp, and an open reading frame of 855 bp encoding a polypeptide of 284 amino acids. Real-time quantitative PCR revealed that α-TM mRNA is initially expressed in unfertilized ovum, indicating the α-TM gene is maternal. Relatively low mRNA levels were observed in different embryonic stages. A higher level of α-TM mRNA was detected 3 days post hatching (dph), while the highest level was measured at 29 dph (metamorphic climax) after which it declined towards the end of metamorphosis. The expression of α-TM mRNA was up-regulated in thyroid hormone-treated larvae at 36 dph, but there was no marked difference at other stages when compared to control animals. After thiourea treatment, the expression of α-TM mRNA declined slightly. These data provide basic information that can be utilized in further studies into the role of α-TM in P. olivaceus development and metamorphosis.
  • 加载中
  • [1] Bing W, Knott A, Redwood C, Esposito G, Purcell I, Watkins H, Marston S. 2000. Effect of hypertrophic cardiomyopathy mutations in human cardiac muscle α-tropomyosin (Asp175Asn and Glu180Gly) on the regulatory properties of human cardiac troponin determined by in vitro motility assay. Journal of Molecular and Cellular Cardiology, 32(8): 1489-1498.
    [2] Fu YS, Shi ZY, Wang GY, Li WJ, Zhang JL, Jia L. 2012. Expression and regulation of miR-1,-133a,-206a, and MRFs by thyroid hormone during larval development in Paralichthys olivaceus. Comparative Biochemistry and Physiology. Part B: Biochemistry & Molecular Biology, 161(3): 226-232.
    [3] Gordon AM, Homsher E, Regnier M. 2000. Regulation of contraction in striated muscle. Physiological Reviews, 80(2): 853-924.
    [4] Inui Y, Miwa S. 1985. Thyroid hormone induces metamorphosis of flounder larvae. General and Comparative Endocrinology, 60(3): 450-454.
    [5] Jagatheesan G, Rajan S, Wieczorek DF. 2010. Investigations into tropomyosin function using mouse models. Journal of Molecular and Cellular Cardiology, 48(5): 893-898.
    [6] Kirwan JP, Hodges RS. 2010. Critical interactions in the stability control region of tropomyosin. Journal of Structural Biology, 170(2): 294-306.
    [7] Li XY, Lin YS, Zhang HW. 2012. Phylogenetic analysis and expression patterns of tropomyosin in amphioxus. Zoological Research, 33(4): 389-394. (in Chinese)
    [8] Marston S, Memo M, Messer A, Papadaki M, Nowak K, McNamara E, Ong R, El-Mezgueldi M, Li XC, Lehman W. 2013. Mutations in repeating structural motifs of tropomyosin cause gain of function in skeletal muscle myopathy patients. Human Molecular Genetics, 22(24): 4978-4987.
    [9] Minami T. 1982. The early life history of a flounder Paralichthys olivaceus [in Wakasa Bay, Japan Sea, Japan]. Bulletin of the Japanese Society of Scientific Fisheries, 48(11):1581-1588.
    [10] Muthuchamy M, Pajsk L, Howles P, Doetschman T, Wieczorek DF.
    [11] 1993. Developmental analysis of tropomyosin gene expression in embryonic stem cells and mouse embryos. Molecular and Cellular Biology, 13(6): 3311-3323.
    [12] Pearlstone JR, Smillie LB. 1983. Effects of troponin-I plus-C on the binding of troponin-T and its fragments to alpha-tropomyosin. Ca2+ sensitivity and cooperativity. Journal of Biological Chemistry, 258(4): 2534-2542.
    [13] Rethinasamy P, Muthuchamy M, Hewett T, Boivin G, Wolska BM, Evans C, Solaro RJ, Wieczorek DF. 1998. Molecular and physiological effects of α-tropomyosin ablation in the mouse. Circulation Research, 82(1): 116-123.
    [14] Schefe JH, Lehmann KE, Buschmann IR, Unger T, Funke-Kaiser H. 2006. Quantitative real-time RT-PCR data analysis: current concepts and the novel “gene expression's CT difference” formula. Journal of Molecular Medicine, 84(11): 901-910.
    [15] Schreiber AM & Specker JL. 1998. Metamorphosis in the Summer Flounder (Paralichthys dentatus): Stage-Specific Developmental Response to Altered Thyroid Status. General and comparative endocrinology, 111(2): 156-166.
    [16] Sumida JP, Wu E, Lehrer SS. 2008. Conserved Asp-137 imparts flexibility to tropomyosin and affects function. Journal of Biological Chemistry, 283(11): 6728-6734.
    [17] Wilbur HM. 1980. Complex life cycles. Annual review of Ecology and Systematics, 11: 67-93.
    [18] Yamano K, Miwa S, Obinata T, Inui Y. 1991. Thyroid hormone regulates developmental changes in muscle during flounder metamorphosis. General and Comparative Endocrinology, 81(3): 464-472.
    [19] Yamano K, Takano-Ohmuro H, Obinata T, Inui Y. 1994. Effect of thyroid hormone on developmental transition of myosin light chains during flounder metamorphosis. General and Comparative Endocrinology, 93(3): 321-326.
    [20] Youson JH. 1988. First metamorphosis. Fish Physiology, 11(Part B): 135-196.
  • 加载中
通讯作者: 陈斌, bchen63@163.com
  • 1. 

    沈阳化工大学材料科学与工程学院 沈阳 110142

  1. 本站搜索
  2. 百度学术搜索
  3. 万方数据库搜索
  4. CNKI搜索

Article Metrics

Article views(493) PDF downloads(1284) Cited by()

Related
Proportional views

cDNA clone and expression analysis of α-Tropomyosin during Japanese flounder (Paralichthys olivaceus) metamorphosis

doi: 10.13918/j.issn.2095-8137.2014.4.307

Abstract: Tropomyosin (TM) plays a critical role in skeletal and cardiac muscle development and function. To assess the functional significance of α-TM in Japanese flounder (Paralichthys olivaceus) development and metamorphosis, cDNA from Japanese flounder was cloned and α-TM mRNA measured during development and metamorphosis. The full-length cDNA is 1 191 bp, including a 5'-untranslated region of 114 bp, a 3'-UTR of 222 bp, and an open reading frame of 855 bp encoding a polypeptide of 284 amino acids. Real-time quantitative PCR revealed that α-TM mRNA is initially expressed in unfertilized ovum, indicating the α-TM gene is maternal. Relatively low mRNA levels were observed in different embryonic stages. A higher level of α-TM mRNA was detected 3 days post hatching (dph), while the highest level was measured at 29 dph (metamorphic climax) after which it declined towards the end of metamorphosis. The expression of α-TM mRNA was up-regulated in thyroid hormone-treated larvae at 36 dph, but there was no marked difference at other stages when compared to control animals. After thiourea treatment, the expression of α-TM mRNA declined slightly. These data provide basic information that can be utilized in further studies into the role of α-TM in P. olivaceus development and metamorphosis.

Hong-Mei ZHANG, Yan-Fang SU, Zhi-Yi SHI, Yuan-Shuai FU. cDNA clone and expression analysis of α-Tropomyosin during Japanese flounder (Paralichthys olivaceus) metamorphosis. Zoological Research, 2014, 35(4): 307-312. doi: 10.13918/j.issn.2095-8137.2014.4.307
Citation: Hong-Mei ZHANG, Yan-Fang SU, Zhi-Yi SHI, Yuan-Shuai FU. cDNA clone and expression analysis of α-Tropomyosin during Japanese flounder (Paralichthys olivaceus) metamorphosis. Zoological Research, 2014, 35(4): 307-312. doi: 10.13918/j.issn.2095-8137.2014.4.307
Reference (20)

Catalog

    /

    DownLoad:  Full-Size Img  PowerPoint
    Return
    Return