생명과학회지 (Journal of Life Science)

  • 제28권3호
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  • Pages.284-292
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  • 2018
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  • 1225-9918(pISSN)
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  • 2287-3406(eISSN)
한국생명과학회 (Korean Society of Life Science)
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양식넙치 멜라닌 농축 호르몬의 특성

Characterization of Melanin-concentrating Hormone from Olive Flounder (Paralichthys olivaceus)

  • 정인영 (국립부경대학교 미생물학과) ;
  • 전정민 (국립부경대학교 자원생물학과) ;
  • 송영환 (국립부경대학교 미생물학과)
  • 투고 : 2017.12.29
  • 심사 : 2018.01.15
  • 발행 : 2018.03.30
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초록

멜라닌 농축 호르몬(melanin-concentrating hormone, MCH)은 17개의 아미노산으로 구성된 환형의 시상하부 펩티드로 색소 침착의 조절인자로서 연어에서 처음 분리되었다. 포유동물의 MCH는 19개의 아미노산으로 구성되어 있으며 섭식 및 에너지 항상성을 조절하는데 관여한다. 본 연구에서는 양식넙치의 다양한 조직에서 MCH 유전자의 발현 분포, 멜라닌 함유 세포의 집적, 포유동물 MCH 수용체와 양식넙치 MCH의 상호작용을 조사하였다. Real-time qPCR을 이용하여 뇌, 정소, 난소에서 MCH 유전자의 발현이 나타나는 것을 확인하였고, 수정 후 발달 단계에서도 MCH 유전자의 발현을 확인할 수 있었다. 합성된 연어 sMCH, 포유류 hMCH, 양식넙치 fMCH, dN-fMCH, dC-fMCH를 양식 넙치의 표피에 처리했을 때 다양한 농도에 따라 멜라닌 함유 세포의 집적이 다양하게 나타났다. 연어 sMCH, 포유류 hMCH에 비해 양식넙치 fMCH의 멜라닌 함유세포의 집적도가 36~99.85%로 비역가를 나타났으나 양식넙치 dN-fMCH, dC-fMCH를 처리한 경우 양식넙치 fMCH에 비해 높은 농도에서 집적이 나타나고 짧은 시간에 분산되었다. 또한, 인간 MCH 수용체와 쥐 MCH 수용체가 발현된 포유동물의 세포주에 양식넙치 fMCH를 처리하여 각 수용체와 결합하는 것을 확인하였다. 이러한 결과는 어류에서 발현되는 MCH가 포유동물의 MCH와 유사한 구조를 가지고 있어 MCH 수용체에 대한 새로운 리간드로서 제공될 수 있으며, 향후 어류의 MCH 수용체에 확대 적용할 수 있을 것이다.

The melanin-concentrating hormone (MCH), a cyclic hypothalamic peptide composed of 17 amino acids, was initially identified in chum salmon (Oncorhynchus keta) as a regulator of pigmentation. Mammalian MCHs are cyclic hypothalamic peptides composed of 19 amino acids that regulate food intake and energy homeostasis. The present study examined not only MCH expression of different tissues but also the melanohore aggregation and intracellular $Ca^{2+}$ influx of fMCH and the other MCH. Real-time qPCR showed that MCH expressed specially in the brain, gonad, and ovary, and expression of MCH was observed during the developmental stages. In the application of synthetic fMCH and both types of synthetic fMCH, dN-fMCH and dC-fMCH, scale melanophore induced significant changes in aggregation activity with various concentrations of MCH. Also, compared to hMCH and sMCH, fMCH exhibited a 36~99.85% increase in relative potency (%), whereas aggregation of dN-fMCH and dC-fMCH remained in a high concentration. However, dispersion was induced rapidly according to be low concentration of dN-fMCH and dC-fMCH. We show that fMCH and its derivates were bound human MCHR1 and rat MCHR expressed in HEK293T cells with nano-molar affinity and are likely to be ligand-induced to mobilize intracellular $Ca^{2+}$. These results may provide new ligands for binding assay with MCHew ligands, as a structure similar to the mammalian MCH structure was discovered in fish. Once the fMCH receptor system is in place, it can be compared to the MCH system of mammals in terms of MCH function.

키워드

참고문헌

  1. Abbott, Caroline, R., Adam, R. K., Alison, M. W., Michela, R., Kevin, G. M., Leighton, J. S., Jeannie, F. T., Mohammad, A. G., Caroline, J. S. and Stephen, R. B. 2003. Identification of hypothalamic nuclei involved in the orexigenic effect of melanin-concentrating hormone. Endocrinology 144, 3943-3949. https://doi.org/10.1210/en.2003-0149
  2. Audinot, V., Beauverger, P., Lahaye, C., Suply, T., Rodriquez, M., Ouvry, C. and Lamamy, V., et al. 2001. Structure-activity relationship studies of melanin-concentrating hormone (MCH)-related peptide ligands at SLC-1, the human MCH receptor. J. Biol. Chem. 276, 13554-13562. https://doi.org/10.1074/jbc.M010727200
  3. Bachner, D., Kreienkamp, H., Weise, C., Buck, F. and Richter, D. 1999. Identification of melanin concentrating hormone (MCH) as the natural ligand for the orphan somatostatin- like receptor 1 (SLC-1). FEBS Lett. 457, 522-524. https://doi.org/10.1016/S0014-5793(99)01092-3
  4. Baker, B. I. 1994. Melanin-concentrating hormone updated: functional considerations. Trends Endocrinol. Metab. 90, 120-126.
  5. Baker, B. I., Eberle, A. N., Baumann, J, B., Siegrist, W. and Girard, J. 1985. Effect of melanin-concentrating hormone on pigment and adrenal cells in vitro. Peptides 6, 1125-1130. https://doi.org/10.1016/0196-9781(85)90438-3
  6. Baker, B., Andrew, L., Len, H. and Stafford, L. 1995. Cloning and expression of melanin-concentrating hormone genes in the rainbow trout brain. Neuroendocrinology 61, 67-76. https://doi.org/10.1159/000126814
  7. Balm, P. H. M. and Diet, G. 1998. The melanin-concentrating hormone system in fish. Ann. N. Y. Acad. Sci. 839, 205-209. https://doi.org/10.1111/j.1749-6632.1998.tb10760.x
  8. Bittencourt, J. C., Presse, F., Arias, C., Peto, C., Vaughan, J., Nahon, J. L., Vale, W. and Sawchenko, P. E. 1992. The melanin-concentrating hormone system of the rat brain: an immuno- and hybridization histochemical characterization. J. Comp. Neurol. 319, 218-245. https://doi.org/10.1002/cne.903190204
  9. Cardinaud, B., Fleur D. T., Jacques, D., Jean, A. B. and Jean, L. N. 2004. Comparative analysis of melanin-concentrating hormone structure and activity in aishes and mammals. Peptides 25, 1623-1632. https://doi.org/10.1016/j.peptides.2004.05.020
  10. Della-Zuana, O., Presse, F., Ortola, C., Duhault, J., Nahon, J. L. and Levens, N. 2002. Acute and chronic administration of melanin-concentrating hormone enhances food intake and body weight in wistar and Sprague-Dawley rats. Int. J. Obes. Relat. Metab. Disord. 26, 1289-1295. https://doi.org/10.1038/sj.ijo.0802079
  11. Gonzalez, M. I., Vaziri, S. and Wilson, C. A. 1996. Behavioral effects of alpha-MSH and MCH after central administration in the female rat. Peptides 17, 171-177. https://doi.org/10.1016/0196-9781(95)02092-6
  12. Griffond, B. and Baker, B. I. 2002. Cell and molecular cell biology of melanin-concentrating hormone. Int. Rev. Cytol. 213, 233-277.
  13. Groneveld, D., Eckhardt, E. R. M., Coenen, A. J. M., Martens, G. J. M., Balm, P. H. M. and Wendelaar Bonga, S. E. 1995. Expression of tilapia prepro- melanin-concentrating hormone mRNA in hypothalamic and neurohypophysial Cells. J. Mol. Endocrinol. 14, 199-207. https://doi.org/10.1677/jme.0.0140199
  14. Groneveld, D., Balm, P. H. M. and Sjoerd, E. W. B. 1996. Melanin-concentrating hormone gene-related peptide stimulates ACTH, but Not ${\alpha}$-MSH, release from the tilapia pituitary. J. Endocrinol. 148, R1-4. https://doi.org/10.1677/joe.0.148R001
  15. Groneveld, D., Balm, P. H. and Wendelaar Bonga, S. E. 1995. Biphasic effect of MCH on ${\alpha}$-MSH release from the tilapia (Oreochromis mossambicus) pituitary. Peptides 16, 945-949. https://doi.org/10.1016/0196-9781(95)00063-P
  16. Jeon, J. M. and Song, Y. H. 2003. Cloning of melanin concentrating hormone cDNA gene from olive flounder (Paralichthys olivaceus). J. Kor. Fishe. Soc. 36, 442-448.
  17. Hiroyuki, I., Kawazoe, I. and Kawauchi, H. 1985. Fish melanin- concentrating hormone disperses melanin in amphibian melanophores. Gen. Comp. Endocrinol. 58, 486-490. https://doi.org/10.1016/0016-6480(85)90123-6
  18. Kawauchi, H. 2006. Functions of melanin-concentrating hormone in fish. J. Exp. Zool. Part A Comp. Exp. Biol. 305, 751-760.
  19. Kawauchi, H. and Baker, B. I. 2004. Melanin-concentrating hormone signaling systems in fish. Peptides 25, 1577-1584. https://doi.org/10.1016/j.peptides.2004.03.025
  20. Lakaye, B., Bernard, C., Sophie, H. and Thierry, G. 2009. Melanin-concentrating hormone and immune function. Peptides 30, 2076-2080. https://doi.org/10.1016/j.peptides.2009.05.004
  21. Ludwig, D. S., Nicholas, A. T., Jason, W. M., Rohit, K., Efi, K., Joel, E., Bradford, L., Jeffrey, S. F. and Eleftheria, M. F. 2001. Melanin-concentrating hormone overexpression in transgenic mice leads to obesity and insulin resistance. J. Clin. Invest. 107, 379-386. https://doi.org/10.1172/JCI10660
  22. Masao, O., Wada, C., Oikawa, I., Kawazoe, I. and Kawauchi, H. 1988. Structures of two kinds of mRNA encoding the chum salmon melanin-concentrating hormone. Gene 71, 433-38. https://doi.org/10.1016/0378-1119(88)90060-1
  23. Minth, C. D., Qiu, H., Akil, H., Watson, S. J. and Dixon, J. E. 1989. Two precursors of melanin-concentrating hormone: DNA sequence analysis and in situ immunochemical localization. Proc. Natl. Acad. Sci. USA. 86, 4292-4296. https://doi.org/10.1073/pnas.86.11.4292
  24. Mizusawa, K., Kawashima, Y., Sunuma, T., Hamamoto, A., Kobayashi, Y., Kodera, Y., Saito, Y. and Takahashi, A. 2015. Involvement of melanin-concentrating hormone 2 in background color adaptation of barfin flounder, Verasper moseri. Gen. Comp. Endocrinol. 214, 140-148. https://doi.org/10.1016/j.ygcen.2014.07.008
  25. Nahon, J. L., Presse, F., Bittencourt, J. C., Sawchenko, P. E. and Vale, W. 1989. The rat melanin-concentrating hormone messenger ribonucleic acid encodes multiple putative neuropeptides coexpressed in the dorsolateral hypothalamus. Endocrinology 125, 2056-65. https://doi.org/10.1210/endo-125-4-2056
  26. Nahon, J. L. and Presset, F., et al. 1991. Identification of a single melanin-concentrating hormone messenger ribonucleic acid in coho salmon: structural relatedness with 7SL ribonucleic acid. J. Neuroendocrinol. 3, 173-183. https://doi.org/10.1111/j.1365-2826.1991.tb00260.x
  27. Parker, J. A. and Bloom, S. R. 2012. Hypothalamic neuropeptides and the regulation of appetite. Neuropharmacology 63, 18-80. https://doi.org/10.1016/j.neuropharm.2012.02.004
  28. Qu, D., David, S. L., Steen, G., Megan, P., Pelleymounter, M. A., Cullen, M. J., Mathes, W. F., Przypek, J., Kanarek, R. and Maratos-Flier, E. 1996. A Role for melanin-concentrating hormone in the central regulation of feeding behaviour. Nature 380, 243-247. https://doi.org/10.1038/380243a0
  29. Rossi, M., Choi, S. J., O'Shea, D., Miyoshi, T., Ghatei, M. A. and Bloom, S. R. 1997. Melanin-concentrating hormone acutely stimulates feeding, but chronic administration has no effect on body weight. Endocrinology 138, 351-355. https://doi.org/10.1210/endo.138.1.4887
  30. Saito, Y., Nothacker, H. P., Wang, Z., Lin, S. H., Leslie, F. and Civelli, O. 1999. Molecular characterization of the melanin- concentrating hormone receptor. Nature 400, 265-269. https://doi.org/10.1038/22321
  31. Sanchez, M., Baker, B. I. and Celis, M. 1997. Melanin-concentrating hormone (MCH) antagonizes the effects of ${\alpha}$-MSH and neuropeptide E-I on grooming and locomotor activities in the rat. Peptides 18, 393-396. https://doi.org/10.1016/S0196-9781(96)00327-0
  32. Shimada, M., Nicholas, A. T., Bradford, B. L., Jeffrey, S. F. and Eleftheria, M. F. 1998. Mice lacking melanin-concentrating hormone are hypophagic and lean. Nature 396, 670-679. https://doi.org/10.1038/25341
  33. Shimomura, Y., Masaaki, M., Tsukasa, S., Yoshihiro, I., Michiko, A., Tsutomu, K., Haruo, O., Osamu, N., Yasuhiro, S. and Masahiko, F. 1999. Isolation and identification of melanin- concentrating hormone as the wndogenous ligand of the SLC-1 receptor. Biochem. Biophys. Res. Commun. 261, 622-626. https://doi.org/10.1006/bbrc.1999.1104
  34. Takahashi, A., Takayoshi, K., Yuki, K. and Takeshi, Y., et al. 2007. The melanin-concentrating hormone receptor 2 (MCH-R2) mediates the effect of MCH to control body color for background adaptation in the barfin flounder. Gen. Comp. Endocrinol. 151, 210-219. https://doi.org/10.1016/j.ygcen.2007.01.011
  35. Takahashi, A., Tsuchiya, K., Yamanome, T., Masafumi, A., Akikazu, Y., Kunio, Y. and Kawauchi, H. 2004. Possible involvement of melanin-concentrating hormone in food intake in a teleost fish, barfin flounder. Peptides 25, 1613-1622. https://doi.org/10.1016/j.peptides.2004.02.022
  36. Tritos, N. A., Vicent, D., Gillette, J., Ludwig, D. S., Flier, E. S. and Maratos-Flier, E. 1998. Functional interactions between melanin-concentrating hormone, neuropeptide Y, and anorectic neuropeptides in the rat hypothalamus. Diabetes 47, 1687-1692.
  37. Vaughan, J. M., Fischer, W. H., Hoeger, C., Rivier, J. and Vale, W. 1989. Characterization of melanin-concentrating hormone from rat hypothalamus. Endocrinology 125, 1660-1665. https://doi.org/10.1210/endo-125-3-1660
  38. Yang, Y. 2011. Structure, function and regulation of the melanocortin receptors. Eur. J. Pharmacol. 660, 125-130. https://doi.org/10.1016/j.ejphar.2010.12.020
  39. Zheng, H., Patterson, L. M., Morrison, C., Banfield, B. W., Randall, J. A., Browning, K. N., Travagli, R. A. and Berthoud, H. R. 2005. Melanin concentrating hormone innervation of caudal brainstem areas involved in gastrointestinal functions and energy balance. Neuroscience 135, 611-625. https://doi.org/10.1016/j.neuroscience.2005.06.055