مطالعه بیان ژن­ های GH و گرلین در دوره تکامل لاروی ماهی گورخری

نویسندگان

1 دانشگاه علوم کشاورزی و منابع طبیعی گرگان،

2 دانشگاه علوم کشاورزی و منابع طبیعی گرگان

چکیده
ماهی گورخری به عنوان یک گونه مهم در علم ژنتیک است و با توجه به نزدیکی ژنوم آن به ژنوم انسان، بررسی بیان برخی از ژن­های مربوط به رشد و اشتها طی دورۀ تکامل لاروی این ماهی ضروری به نظر می­رسد. ژن­های کدکننده هورمون رشد و اشتها (GH و گرلین) در سنتز و رهاسازی هورمون رشد نقش دارند که می‌توانند به عنوان ژن­های اقتصادی در ماهیان مطرح شوند. با توجه به اهمیت ژن­های مذکور در دوران تکامل لاروی، تحقیق حاضر به ­منظور بررسی روند فعالیت آنها در مراحل ابتدایی رشد صورت گرفت. نمونه ­برداری از لاروها در روزهای 4، 7، 10، 15، 30 و 45 پس از تفریخ صورت پذیرفت. نمونه­ ها بلافاصله در ازت مایع قرار گرفتند و تا زمان استخراج RNA (با استفاده از کیت RNX-Plus) در فریزر 80- درجۀ سانتی­گراد نگهداری شدند. داده ­های ژن­های هدف با استفاده از ژن رفرنس β-actin به روش ∆∆CT- 2 تحلیل ­شد. بیان ژن­های مذکور در طی مراحل مختلف تکاملی تفاوت معنی­ داری با یکدیگر داشتند؛ به­ طوری که بیان ژن GH و گرلین به­ترتیب در روز 4 و 10 پس از تفریخ به­ طور معنی­ داری بیشتر از سایر روزهای نمونه ­برداری بود (P<0.05). بطور کلی با توجه به اهمیت بسیار زیاد ژن­های مرتبط با رشد و اشتها که تأثیر به سزایی در روند بازمانی و بقاء لاروها در مراحل اولیه زندگی بر عهده دارند، در مراحل اولیۀ لاروی و روزهای ابتدایی لاروها بیان ژن­های GH و گرلین، بیشتر از سایر دوره ­های تکاملی است.

کلیدواژه‌ها


عنوان مقاله English

Study of GH and Ghrelin genes expression during the larvae developmental period in Danio rerio

نویسندگان English

Hamed Paknejad 1
Tayebeh Enayat Gholampour 1
Roghayeh Safari 2
Seyed Hossein Hossenifar 1
1 Gorgon University of Agricultural Science and Natural Resources, Gorgan, Iran
2 Gorgon University of Agricultural Science and Natural Resources, Gorgan, Iran
چکیده English

Zebra fish is an important species in genetics and considering the proximity of its genome to the human genome, investigating the expression of some of the growth and appetite genes during its larvae development is essential. Genes coding growth and appetite (GH and ghrelin) hormones are involved in the synthesis and release of growth hormone, which can be considered to be economic genes in pisciculture. Given the importance of these genes during the early larvae development stages, this study was performed to assess their activity. Samples were collected at 4, 7, 10, 15, 30 and 45 post-hatching days. Samples were immediately placed in liquid nitrogen (-196 degree centigrade) and then stored in a freezer at -80 degree centigrade until RNA extraction (using RNX-Plus kit). To analyze normal expression of target genes, reference gene β-actin was used by 2-∆∆Ct method. The expression of genes associated with the growth and appetite was significantly different at various stages of the development of zebra fish, as the gene expression of GH on day 4 and ghrelin gene on day 10 after hatching were significantly higher compared with other samples (P<0.05). Overall, the expression of GH and ghrelin genes at the early stages of growth is higher than other fish developmental stages due to their great importance in the survival of larvae at these stages of life.



<span style="mso-ascii-font-family: " times="" new="" roman";mso-ascii-theme-font:major-bidi;mso-hansi-font-family:"times="" roman";="" mso-hansi-theme-font:major-bidi;mso-bidi-font-family:"times="" mso-bidi-theme-font:major-bidi;color:#222222"="">

کلیدواژه‌ها English

appetite
beta actin
genetic
Growth Hormone
ontogenyrio
Barros, T.P., Alderton, W.K., Reynolds, H.M., Roach, A.G. and Berghmans, S. 2008. Zebrafish: an emerging technology for in vivo pharmacological assessment to identify potential safety liabilities in early drug discovery. – Brit. J. Pharm. 154: 1400-1413.
Bustin, A.S., Benes, V., Garson, J.A., Healmans, J., Huggett, J., Kubista, M., Muller, R., Nolaan, T., Pfaffl, M., Shipley, G., Vandesompele, J. and Wittwer, C.T. 2009. The MIQE guidelines: minimum information for publication of quantitative real time PCR experiments. – Clin. Chem. 55: 611-622.
Canestro, C., Yokoi, H. and Postlethwait, J. 2007. Evolutionary developmental biology and geno-mics. – Nat. Rev. Gen. 8: 932-942.
Cheshenko, K., Brion, F., Page, Y., Hinfray, N., Pakdel, F., Kah, O., Segner, H. and Eggen, P. 2007. Expression of Zebra fish aromatase cyp19a and cyp19bm genes in response to the ligands of estrogen receptor and aryl hydrocarbon receptor. – Toxi. Sci. 96: 255-267.
Grunwald, D. and Eisen, J. 2002. Headwaters of the zebrafish-emergence of a new model vertebrate. – Nat. Rev. Gen. 3: 717-724.
Hill, A.J., Teraoka, H., Heideman, W. and Peterson, R.E. 2005. Zebrafish as a model vertebrate for investigating chemical toxicity. – Toxi. Sci. 86: 6-19.
Kaiya, H., Kojima, M., Hosoda, H., Riley, L.G., Hirano, T., Grau, E.G. and Kangawa, K. 2003a. Amidated fish ghrelin: purification, cDNA cloningin the Japanese eel and its biological activity. – J. Endoc. 176: 415-423.
Kaiya, H., Kojima, M., Hosoda, S., Moriyama, M., Takahashi, A. and Kawauchi, H. 2003b. Peptide purification, cDNA and genomic DNA cloning, and functional characteristics of ghrelin in rainbow trout. – J. Endoc. 144: 5215-5226.
Kawakoshi, A., Kaiya, H., Riley, L.G., Hirano, T., Grau, E.G., Miyazato, M., Hosoda, H. and Kangawa, K. 2007. Identification of a ghrelin-like peptide in two species of shark, Sphyrna lewini and Carcharhinus melanopterus. – Gen. Com. Endoc. 151: 259-268.
Kojima, M., Hosoda, H. and Kangawa, K. 2001. Purification and distribution of ghrelin: the natural endogenous ligand for the growth hormone secretagogue receptor. – Horm. Res. 56: 93-97.
Kolangi Miandare, H., Farahmand, H., Akbarzadeh, A., Ramezanpour, S., Kaiya, H., Miyazato, M. and Nikinma, M. 2013. Developmental transcription of genes putatively associated with growth in two sturgeon species of different growth rate. – Gen. Comp. Endoc. 182: 41-47.
Larionov, A., Krause, A. and Miller, W. 2005. A standard curve based method for relative real time PCR data processing. BMC Bioinformatics 6: 62-77.
Lawrence, C. 2011. Advances in Zebrafish husbandry and management. – Methods Cell Biol. 104: 429-45.
Lawrence, C. 2011. Advances in zebrafish husbandry and management. – Methods Cell Biol. 104: 429-451.
Lee, L.T.O., Nong, G. and Chan Y.H .2001. Molecular cloning of a teleost growth hormone recept or and its funct ional interaction with human grow the hormone. – Gene. 270: 121-129.
Lieschke, G.J. and Currie, P.D. 2007. Animal models of human disease: zebrafish swim into view. – Natu. Rev. Genet. 5: 353-367.
Livak, K.J. and Schmittgen, T.D. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. – Methods 25: 402-408.
Parhar, I.S., Sato, H. and Sakuma, Y. 2003. Ghrelin gene in cichlid fish is modulated by sex and development. – Biochem. Biophys. Res. Communi. 305: 169-175.
Radonic, A., Thulke, S., Mackay, I.M., Landt, O., Siegert, W. and Nitsche A. 2004. Guideline to reference gene selection for quantitative real-time PCR. Biochem. Biophys. Res. Commun. 313: 856-862.
Riley, L.G., Hirano, T. and Grau, E.G. 2002. Rat ghrelin stimulates growth hormone and prolactin release in the tilapia, Oreochromis mossambicus. – Zool. Sci. 19: 797-800.
Schmid, A.C., Lutz, I., Kloas, W. and Reinecke, M. 2003. Thyroid hormone stimulates hepatic IGF-I mRNA expression in a bony fish, the tilapia, Oreochromis mossambicus, in vitro and in vivo. – Gen. Comp. Endoc. 130: 129-134.
Scholz, S., Fischer S., Gundel, U., Kuster, E., Luckenbach, T. and Voelker, D. 2008. The zebrafish embryo model in environmental risk assessment applications beyond acute toxicity testing. – Environ. Sci. Pollut. Res. 15: 394-404
Shepherd, B.S., Johnson, J.K., Silverstein, J.T., Parhar, IS., Vijayan, M.M., McGuire, A. and Weber, G.M. 2007. Endocrine and orexigenic actions of growth hormone secretagogues in rainbow trout (Oncorhynchus mykiss). – Comp. Biochem. Phys. A. 146: 390-399.
Spence, R., Gerlach, G., Lawrence, C. and Smith, C. 2008. The behavior and ecology of the zebrafish, Danio rerio. – Biol. Rev. 83: 13-34.
Suda, A., Kaiyai, H., Nikaido, S., Moshiro, H. and Ando, K. 2012. Identification and gene expression analysis of ghrelin in the stomach pacific Bluefin tuna (Thunnus orientalis). – Gen. Comp. Endoc. 178: 89-97.
Torgersen, J., Nourizadeh-Lillabadi, R., Husebye, H. and Alestrom, P. 2002. In silico and in situ characterization of the zebrafish (Danio rerio) Gnrh3 (sGnRH) gene. – BMC Genom. 3: 25-36.
Trant, J., Gavasso, S., Ackers, J., Chung, B. and Place, A. 2001. Developmental expression of cytochrome P450 aromatase genes (CYP19a and CYP19b) in Zebrafish fry (Danio rerio). – J. Exper. Zool. 290: 475-483.
Unniappan, S., Canosa, L.F. and Peter, R.E. 2014. Orexigenic activities of ghrelin in goldfish: feeding-induced vagaries in brain and gut mRNA appearance and serum levels, and comebacks to central and peripheral injections. – Physiol. Rev. 12: 38-50.
Van der Lely, A.J., Tschöp, M., Heiman, M.L. and Ghigo, E. 2004. Biological, physiological, pathophysiological and pharmacological aspects of ghrelin. – Endocrin. Rev. 25: 426-457.
Yeung, C.M., Chan, C.B., and Cheng, C.H. 2004. Isolation and characterization of the 5'-flanking region of the growth hormone secretagogue receptor gene from black seabream Acanthopagrus schlegeli. – Mol. Cell. Endocrin. 223: 5-15.
دوره 6، شماره 2 - شماره پیاپی 20
تابستان 1398
صفحه 148-154

  • تاریخ دریافت 18 خرداد 1405
  • تاریخ انتشار 18 خرداد 1405