بررسی بیان ژنی تنظیم‌کننده‌های کلیدی تشکیل لایه‌های زاینده در جنین‌های جوجه با استفاده از تکنیک RT-PCR

نوع مقاله : مقاله پژوهشی

نویسندگان

1 استادیار، دانشکده دامپزشکی، دانشگاه بصره، بصره، عراق.

2 مدرس، دانشکده دامپزشکی، دانشگاه بصره، بصره، عراق

چکیده

هدف: مطالعات مختلف نشان داده‌اند که ژن PRDM14 نقش بسیار مهم و بنیادی در رشد اولیه جنین جوجه و تشکیل لایه‌های زاینده دارد. هدف این مطالعه بررسی الگوی بیان ژن PRDM14 در مراحل اولیه تکوین جنینی و همچنین بررسی نقش تکاملی آن بود. این پژوهش همچنین به دنبال ساده‌سازی شناسایی خانواده‌های اصلی فاکتورهای رونویسی فعال در پنج روز نخست رشد جنینی است.
مواد و روش‌ها: برای مقایسه PRDM14 با سایر اعضای خانواده PRDM، از تحلیل فیلوژنتیک استفاده شد. همچنین، پروفایل‌گذاری مقایسه‌ای بیان ژن برای بررسی فعالیت ژنی در مراحل E1 تا E5 انجام شد که نمایانگر اولین و حساس‌ترین مراحل رشد جنین جوجه هستند. از مجموعه‌ داده‌های عمومی ژنومی و خانواده‌های ژنی حاشیه‌نویسی‌شده برای شناسایی فاکتورهای رونویسی و تعیین توزیع آن‌ها در سراسر ژنوم استفاده شد.
نتایج: تحلیل‌ها نشان داد که ژن PRDM14 و سایر ژن‌های خانواده PRDM الگوی واگرایی تکاملی مشابهی با سایر مهره‌داران دارند. مراحل E1 تا E5 به‌عنوان حیاتی‌ترین مراحل تکوین جنینی شناسایی شدند و تغییرات عمده‌ای در بیان ژن در این دوره‌ها مشاهده شد. در بین جنین‌های بررسی‌شده، 1097 فاکتور رونویسی شناسایی شد. این ژن‌ها دارای توزیع تصادفی نبودند، بلکه الگوی ساختاری ژنومی معناداری داشتند. خانواده‌های فاکتور رونویسی مانند bHLH، Zf-C2H2 و Homeobox نیز شناسایی شدند که نقش مهمی در رشد اولیه دارند. پروتئین‌های هومئوباکس از ۶۰ اسید آمینه تشکیل شده‌اند و برای تنظیم بسیاری از فرآیندهای تکوینی ضروری هستند. فاکتورهای رونویسی ISL1، MKX، NANOG و CDX2 شناسایی و در گروه‌های Homeobox و bHLH طبقه‌بندی شدند. این فاکتورها در تعیین سرنوشت سلولی، تشکیل دودمان‌های سوماتیک و پلوری‌پوتنسی نقش دارند. در روزهای ۴ و ۵، گروه bHLH بیان بالایی نشان داد که نقش اساسی در تکوین دستگاه عصبی و سلول‌های زایشی دارد.
نتیجه‌گیری: این مطالعه نشان می‌دهد که مراحل اولیه تکوین جنینی شامل فعالیت هماهنگ خانواده‌های اصلی فاکتورهای رونویسی است. الگوی بیان ساختاریافته و فعال‌سازی زودهنگام PRDM14 نشان‌دهنده نقش اساسی آن در هدایت هویت سلولی و مسیرهای تکوینی در جنین‌های جوجه است.

کلیدواژه‌ها

عنوان مقاله [English]

Gene expression study of key regulators of germ layer formation in chicken embryos using RT-PCR technique

نویسندگان [English]

  • Rana Khalaf Abdulsamad 1
  • Ashwaq Raheem Nazzal 1
  • Zainab Abbas Musa 2
1 Assistant professor, College of Veterinary Medicine, University of Basra, Basra, Iraq
2 Assistant Lecture, College of Veterinary Medicine, University of Basra, Basra, Iraq.
چکیده [English]

Objective
Various studies have shown that the PRDM14 gene plays a very important and fundamental role in the early development of the chicken embryo and the formation of the germ layer. The aim of this study was to investigate the expression pattern of the PRDM14 gene in early embryogenesis and also to investigate its evolutionary role. This work also seeks to simplify the identification of major transcription factor families active in the first five days of development.

Materials and methods
For comparison of PRDM14 and other members of the PRDM family, we used a phylogenetic analysis. Comparative expression profiling was also conducted to track gene activity during stages E1-E5, which represent the earliest and most sensitive phases of chicken embryonic development. Public genomic datasets and annotated gene families were used to identify transcription factors and determine their distribution across the genome.

Results
The analysis showed that PRDM14 and other PRDM family genes follow an evolutionary divergence pattern similar to that of other vertebrates. The stages E1-E5 were identified as the most crucial in embryogenesis, showing major shifts in gene expression. Among the studied chicken embryos, 1097 transcription factors were observed. These genes did not have random distribution, but they have significant genomic structure pattern. Transcription factor families such as bHLH, Zf-C2H2, and Homeobox were also detected in this study. They play important role in early development. 60 amino acids make up Homeobox proteins. These proteins are essential for regulating many developmental processes. ISL1, MKX, NANOG, and CDX2 are key transcription factors identified and classified into the Homeobox and bHLH groups. These factors are involved in cell fate determination, somatic lineage formation, and pluripotency. At days 4 and 5, the bHLH group of transcription factors showed high expression. These factors were found to play an important and essential role in the nervous system and germ cell development.

Conclusion
This study shows that the early stages of embryogenesis involve coordinated activity of major transcription factor families. The structured expression pattern and early activation of PRDM14 suggest its essential role in guiding cell identity and developmental pathways in chicken embryos.

کلیدواژه‌ها [English]

  • chicken embryos
  • embryonic development
  • germ cells
  • Homeobox
  • PRDM14

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مجله بیوتکنولوژی کشاورزی
دوره 18، شماره 1
فروردین 1405
صفحه 241-258

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