بررسی بیان ژن لیزوزیم نوع-C در بافت‌های قدامی کلیه، کبد، پانکراس و طحال مولدین ماهی قره برون (Acipenser persicus Borodin, 1897) با استفاده از روش real time RT-PCR

نوع مقاله: مقاله کامل

نویسندگان

1 دانش‌آموخته کارشناسی ارشد شیلات، دانشکده کشاورزی و منابع طبیعی، دانشگاه گنبدکاووس، گلستان، ایران

2 استادیار گروه شیلات، دانشکده کشاورزی و منابع طبیعی، دانشگاه گنبدکاووس، گلستان، ایران

3 گروه بهداشت عمومی و مواد غذایی، دانشکده دامپزشکی، دانشگاه شهید باهنرکرمان، کرمان ایران

4 دانشیار گروه شیلات، دانشکده کشاورزی و منابع طبیعی، دانشگاه گنبدکاووس، گلستان، ایران

چکیده

لیزوزیم‌ها از مهم‌ترین پروتئین‌های سیستم ایمنی ذاتی در ماهیان به‌عنوان یک پپتید ضد باکتریایی بخصوص در برابر باکتری‌های گرم مثبت شناخته می‌شوند. با توجه به قابلیت انتقال این آنزیم از مولدین به نسل بعد و همچنین کسب پیشرفت‌های اخیر درزمینه ژنتیک مولکولی در استفاده از روش‌های نوین درزمینه آنالیز mRNA با استفاده از روش Real Time PCR جهت اندازه‌گیری میزان بیان ژن‌های مختلف، مطالعه حاضر باهدف ارزیابی میزان بیان ژن لیزوزیم نوع-C در بافت‌های کبد، قدامی کلیه، پانکراس و طحال در ماهیان مولد قره برون (Acipenser persicus) با استفاده از روش مذکور مورد بررسی قرار گرفت. بدین منظور نمونه‌برداری از بافت‌های موردنظر در زمان تکثیر مصنوعی این‌گونه، پس از انجام عملیات تخم‌کشی، اسپرم‌گیری و کشتار از 16 عدد ماهی مولد (هشت عدد ماهی نر، هشت عدد ماهی ماده) انجام شد. RNA کل استخراج و سنتز cDNA از بافت‌های تفکیک‌شده انجام شد. در ادامه بیان mRNA ژن لیزوزیم نوع-C در بافت‌های مختلف با استفاده از تکنیک real time RT- PCR به‌وسیله ژن β-actin به‌عنوان کنترل داخلی مورد ارزیابی قرار گرفت. نتایج به‌دست‌آمده نشان داد ژن لیزوزیم نوع-C در تمام بافت‌های تحت بررسی بیان دارد و حداکثر تعداد نسخه‌های mRNA این ژن در بافت طحال و کمترین تعداد آن در بافت پانکراس مشاهده شد (01/0 > p). درمجموع، با توجه به نقش عوامل مختلف مانند جنسیت، سن، اندازه، فصل، دمای آب، pH، مسمومیت‌ها، عفونت‌ها و درجه عوامل استرس‌زا در میزان فعالیت و یا سطوح آنزیم لیزوزیم که درنهایت ممکن است باعث فعال شدن مستقیم رونویسی ژن لیزوزیم گردند و اهمیت ماهیان مولد خاویاری می‌توان بیان نمود که این ژن به‌خوبی می‌تواند به‌عنوان یک ژن مرجع به‌منظور پیش‌آگاهی، شناسایی و غربالگری ماهیان مولد بیمار و درمان آن‌ها و کنترل محیط‌زیست در این‌گونه مورداستفاده قرار گیرد.

کلیدواژه‌ها

موضوعات


عنوان مقاله [English]

Investigation of the expressions of the C-type Lysozyme gene in head kidney, liver, pancreas and spleen tissues in persian breeding sturgeon (Acipenser persicus Borodin, 1897) by using real time RT- PCR assays

نویسندگان [English]

  • M.R. Bivareh 1
  • H. Gholipour Kannani 2
  • B. Sadeghi 3
  • H. Jafaryan 4
1 Department of Fisheries and Forestry, Faculty of natural resource, Gonbad Kavous University, Golestan, Iran.
2 Assistant professor, Department of Fisheries and Forestry, Faculty of natural resource, Gonbad Kavous University, Golestan,Iran.
3 Food Hygiene and Public Health Department, Faculty of Veterinary Medicine, Shahid Bahonar University of Kerman, Kerman.
4 Associate professor, Department of Fisheries and Forestry, Faculty of natural resource, Gonbad Kavous University, Golestan, Iran.
چکیده [English]

Lysozymes are important proteins of the innate immune system for the defense against bacterial infection, especially gram-positive bacteria in fishes. According to the ability to transfer this gene from breeders to the next generation, as well as recent advantage-ground in molecular genetics on applying novel methods in the field of mRNA analysis using real time RT-PCR to measure the expression of the different genes, the present study was conducted to evaluate the expression of the C- type lysozyme gene in tissues of liver, head of kidney, pancreas and spleen in breeding of Acipenser persicus. For this purpose, tissue sampling was done after artificial breeding from 16 fish (8 males and 8 females). The tissues were transported to the laboratory in liquid nitrogen. Total RNA was isolated and cDNA synthesis was performed from resected tissues. The mRNA expression of the C-type Lysozyme gene in different tissues were detected by real time quantitative PCR with β-actin gene as an internal control. Expression of the c-type lysozyme gene was detected in all tested tissues. The maximum copy numbers of the mRNA expression of the C-type Lysozyme gene were detected in spleen tissue and the lowest expression of this enzyme was determined in the pancreas tissue (p < 0.01). Overall, considering the role of different factors such as sex, age, size, season, water temperature, pH, toxicants, infections and type of stressors in the lysozyme activity or level, which may have eventually caused the direct activation of transcription of lysozyme gene, as well as considering the importance of breeding of sturgeons, we could claim that this gene can well be used as a reference gene for the prognosis and screening of immune deficiency and control of their environment in A. persicus.

کلیدواژه‌ها [English]

  • Gene expression
  • Lysozyme-C
  • real time RT-PCR
  • Acipenser persicus

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