Journal of Mazandaran University of Medical Sciences
مجله دانشگاه علوم پزشکی مازندران
J Mazandaran Univ Med Sci
Medical Sciences
http://jmums.mazums.ac.ir
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2022
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مقایسه میزان بیان آلکالین فسفاتاز، ژن Gfra1، Lin28 و Sall4 در سلولهای بنیادی جنینی، سلولهای بنیادی اسپرم ساز و سلولهای شبه بنیادی جنینی در موش
Comparing the Expression Levels of Alkaline Phosphatase, Gfra1, Lin28, and Sall4 Genes in Embryonic Stem Cells, Spermatogonial Stem Cells, and Embryonic Stem-Like Cells in Mice
بيولوژي
Biology
پژوهشي-کامل
Research(Original)
<span style="font-size:12pt"><span style="line-height:95%"><span style="direction:rtl"><span style="unicode-bidi:embed"><span style="font-family:"Times New Roman","serif""><span lang="FA" style="line-height:95%"><span style="font-family:"B Koodak""><span style="color:black"><span style="letter-spacing:-.1pt">سابقه</span></span></span></span><span lang="AR-SA" style="line-height:95%"><span style="font-family:"B Koodak""><span style="color:black"><span style="letter-spacing:-.1pt"> و هدف:</span></span></span></span> <a name="_Hlk101604431"><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar"">اسپرماتوژنز یک فرآیند سازمان</span></span></a><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar"">یافته است که فاکتورهای فراوانی بر</span></span><i> </i><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar"">کنترل و تنظیم آن اثرگذار هستند. آلکالین فسفاتاز به همراه ژن</span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar"">های </span></span><i><span dir="LTR" style="font-size:10.0pt"><span style="line-height:95%">Gfra1</span></span></i><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar"">، </span></span><i><span dir="LTR" style="font-size:10.0pt"><span style="line-height:95%">Lin28</span></span></i> <span lang="FA" style="line-height:95%"><span style="font-family:"B Zar"">و </span></span><i><span dir="LTR" style="font-size:10.0pt"><span style="line-height:95%">Sall4</span></span></i> <span lang="FA" style="line-height:95%"><span style="font-family:"B Zar"">از جمله فاکتورهای اصلی دخیل در این فرآیند هستند که با یکدیگر در ارتباط هستند.</span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""> هدف از انجام این مطالعه بررسی و مقایسه میزان بیان آنزیم آلکالین فسفاتاز و همچنین ژن</span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar"">های</span></span><i><span dir="LTR" style="font-size:10.0pt"><span style="line-height:95%"> Gfra1</span></span></i><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar"">، </span></span><i><span dir="LTR" style="font-size:10.0pt"><span style="line-height:95%">Lin28</span></span></i> <span lang="FA" style="line-height:95%"><span style="font-family:"B Zar"">و </span></span><i><span dir="LTR" style="font-size:10.0pt"><span style="line-height:95%">Sall4</span></span></i> <span lang="FA" style="line-height:95%"><span style="font-family:"B Zar"">در سلول</span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar"">های بنیادی جنینی، اسپرم</span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar"">ساز، و شبه جنینی در موش می</span></span><span lang="FA" style="line-height:95%"><span style="font-family:"Arial","sans-serif""></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar"">باشد.</span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="color:#0e101a"></span></span></span></span></span></span></span></span><br>
<span style="font-size:12pt"><span style="text-justify:kashida"><span style="text-kashida:0%"><span style="line-height:95%"><span style="direction:rtl"><span style="unicode-bidi:embed"><span style="font-family:"Times New Roman","serif""><span lang="AR-SA" style="line-height:95%"><span style="font-family:"B Koodak""><span style="color:black"><span style="letter-spacing:-.1pt">مواد و روش</span></span></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black"><span style="letter-spacing:-.1pt"></span></span></span></span><span lang="AR-SA" style="line-height:95%"><span style="font-family:"B Koodak""><span style="color:black"><span style="letter-spacing:-.1pt">ها:</span></span></span></span> <span lang="FA" style="line-height:95%"><span style="font-family:"B Zar"">ابتدا سلولهای بنیادی اسپرمساز جداسازی شدند و سپس سلولهای بنیادی جنینی موش و سلولهای </span></span><span dir="LTR" style="font-size:10.0pt"><span style="line-height:95%">ES-like</span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""> تهیه شدند. در مرحله بعدی سنجش بیان آلکالین فسفاتاز با استفاده از رنگ آمیزی آلکالین فسفاتاز بررسی</span></span><span lang="FA" style="line-height:95%"><span style="font-family:"Arial","sans-serif""></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar"">شد. سپس بررسی بیوانفورماتیکی ژن</span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar"">های دارای ارتباط مستقیم انجام شد و برای سنجش بیان ژنهای </span></span><i><span dir="LTR" style="font-size:10.0pt"><span style="line-height:95%">Gfra1</span></span></i><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar"">، </span></span><i><span dir="LTR" style="font-size:10.0pt"><span style="line-height:95%">Lin28</span></span></i><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""> و </span></span><i><span dir="LTR" style="font-size:10.0pt"><span style="line-height:95%">Sall4</span></span></i><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""> از </span></span><i><span dir="LTR" style="font-size:10.0pt"><span style="line-height:95%">Fluidigm PCR</span></span></i><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""> و همچنین از ایمونوهیستوشیمی، برای بیان </span></span><i><span dir="LTR" style="font-size:10.0pt"><span style="line-height:95%">Gfra1</span></span></i><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar"">، استفاده شد.</span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black"></span></span></span></span></span></span></span></span></span></span><br>
<span style="font-size:12pt"><span style="text-justify:kashida"><span style="text-kashida:0%"><span style="line-height:95%"><span style="direction:rtl"><span style="unicode-bidi:embed"><span style="font-family:"Times New Roman","serif""><span lang="AR-SA" style="line-height:95%"><span style="font-family:"B Koodak""><span style="color:black"><span style="letter-spacing:-.2pt">یافته</span></span></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black"><span style="letter-spacing:-.2pt"></span></span></span></span><span lang="AR-SA" style="line-height:95%"><span style="font-family:"B Koodak""><span style="color:black"><span style="letter-spacing:-.2pt">ها:</span></span></span></span> <span lang="FA" style="line-height:95%"><span style="font-family:"B Zar"">ژن</span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar"">های </span></span><i><span dir="LTR" style="font-size:10.0pt"><span style="line-height:95%">Gfra1</span></span></i><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar"">، </span></span><i><span dir="LTR" style="font-size:10.0pt"><span style="line-height:95%">Lin28</span></span></i><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""> و </span></span><i><span dir="LTR" style="font-size:10.0pt"><span style="line-height:95%">Sall4</span></span></i><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""> بیان مثبت و معنی</span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar"">دار (0/05</span></span><span dir="LTR" style="font-size:10.0pt"><span style="line-height:95%">P<</span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar"">) در سلول</span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar"">های بنیادی پرتوان و سلول</span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar"">های جنسی داشتند. آلکالین فسفاتاز نیز دارای بیان مثبت در سلول</span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar"">های بنیادی جنسی و سلول</span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar"">های بنیادی پرتوان بود و در <span style="letter-spacing:-.2pt">سلول</span></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="letter-spacing:-.2pt"></span></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="letter-spacing:-.2pt">های سرتولی و فیبروبلاست</span></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="letter-spacing:-.2pt"></span></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="letter-spacing:-.2pt">ها بیان کم و منفی داشت، که نشان از اختصاصی بودن این آنزیم در این رده</span></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="letter-spacing:-.2pt"></span></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="letter-spacing:-.2pt">های سلولی دارد.</span></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black"><span style="letter-spacing:-.2pt"></span></span></span></span></span></span></span></span></span></span></span><br>
<span style="font-size:12pt"><span style="text-justify:kashida"><span style="text-kashida:0%"><span style="line-height:95%"><span style="direction:rtl"><span style="unicode-bidi:embed"><span style="font-family:"Times New Roman","serif""><span lang="AR-SA" style="line-height:95%"><span style="font-family:"B Koodak""><span style="color:black"><span style="letter-spacing:-.2pt">استنتاج:</span></span></span></span> <span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="letter-spacing:-.2pt">آلکالین فسفاتاز به همراه ژن</span></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="letter-spacing:-.2pt"></span></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="letter-spacing:-.2pt">های </span></span></span><i><span dir="LTR" style="font-size:10.0pt"><span style="line-height:95%"><span style="letter-spacing:-.2pt">Gfra1</span></span></span></i><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="letter-spacing:-.2pt">، </span></span></span><i><span dir="LTR" style="font-size:10.0pt"><span style="line-height:95%"><span style="letter-spacing:-.2pt">Lin28</span></span></span></i><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="letter-spacing:-.2pt"> و </span></span></span><i><span dir="LTR" style="font-size:10.0pt"><span style="line-height:95%"><span style="letter-spacing:-.2pt">Sall4</span></span></span></i><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="letter-spacing:-.2pt">، با توجه به ارتباط شبکه</span></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="letter-spacing:-.2pt">ای با یکدیگر و بیان آن</span></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="letter-spacing:-.2pt">ها در سلول</span></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="letter-spacing:-.2pt">های بنیادی پرتوان و رده زایا، می</span></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="letter-spacing:-.2pt">توانند به عنوان فاکتورهای تشخیصی برای این رده</span></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="letter-spacing:-.2pt">های سلولی استفاده شوند.</span></span></span><span lang="FA" style="line-height:95%"><span style="font-family:"B Zar""><span style="letter-spacing:-.2pt"></span></span></span></span></span></span></span></span></span></span><br>
<span style="font-size:12pt"><span style="line-height:95%"><span style="tab-stops:284.5pt"><span style="direction:rtl"><span style="unicode-bidi:embed"><span style="font-family:"Times New Roman","serif""><span lang="AR-SA" style="font-size:7.0pt"><span style="line-height:95%"><span style="font-family:"B Zar""><span style="color:black"></span></span></span></span></span></span></span></span></span></span><br>
<span style="font-size:12pt"><span style="line-height:150%"><span style="font-family:"Times New Roman","serif""><b><span style="font-size:10.0pt"><span style="line-height:150%"><span style="font-family:"Arial","sans-serif""><span style="letter-spacing:-.2pt">Background and purpose:</span></span></span></span></b> <span style="font-size:10.0pt"><span style="line-height:150%">Spermatogenesis is a well-organized process that is influenced by a variety of factors. Alkaline phosphatase, and </span></span><i><span style="font-size:10.0pt"><span style="line-height:150%">Gfra1</span></span></i><span style="font-size:10.0pt"><span style="line-height:150%">, </span></span><i><span style="font-size:10.0pt"><span style="line-height:150%">Lin28</span></span></i><span style="font-size:10.0pt"><span style="line-height:150%">, and </span></span><i><span style="font-size:10.0pt"><span style="line-height:150%">Sall4 g</span></span></i><span style="font-size:10.0pt"><span style="line-height:150%">enes are among the key players in this interconnected process. This study aimed to investigate</span></span><span style="font-size:10.0pt"><span style="line-height:150%"> the expression levels of </span></span><i><span style="font-size:10.0pt"><span style="line-height:150%">Gfra1</span></span></i><span style="font-size:10.0pt"><span style="line-height:150%">, </span></span><i><span style="font-size:10.0pt"><span style="line-height:150%">Lin28</span></span></i><span style="font-size:10.0pt"><span style="line-height:150%">, and </span></span><i><span style="font-size:10.0pt"><span style="line-height:150%">Sall4</span></span></i><span style="font-size:10.0pt"><span style="line-height:150%"> genes in embryonic, spermatogonial, and embryonic stem<i>-</i>like (ES-like) cells in mice.</span></span></span></span></span><br>
<span style="font-size:12pt"><span style="line-height:150%"><span style="font-family:"Times New Roman","serif""><b><span style="font-size:10.0pt"><span style="line-height:150%"><span style="font-family:"Arial","sans-serif"">Materials and methods:</span></span></span></b><b> </b><span style="font-size:10.0pt"><span style="line-height:150%">First, spermatogonial stem cells were isolated, and then mouse embryonic stem cells and ES-like cells were prepared. Alkaline phosphatase expression was determined by alkaline phosphatase staining. After that, a bioinformatics analysis of directly related genes was done<i>.</i> Then the expression levels of the </span></span><i><span style="font-size:10.0pt"><span style="line-height:150%">Gfra1</span></span></i><span style="font-size:10.0pt"><span style="line-height:150%">, </span></span><i><span style="font-size:10.0pt"><span style="line-height:150%">Lin28</span></span></i><span style="font-size:10.0pt"><span style="line-height:150%">, and </span></span><i><span style="font-size:10.0pt"><span style="line-height:150%">Sall4</span></span></i><span style="font-size:10.0pt"><span style="line-height:150%"> genes were measured using Fluidigm PCR<i>,</i> and immunohistochemistry was done for </span></span><i><span style="font-size:10.0pt"><span style="line-height:150%">Gfra1</span></span></i><span style="font-size:10.0pt"><span style="line-height:150%"> expression.</span></span></span></span></span><br>
<span style="font-size:12pt"><span style="line-height:150%"><span style="font-family:"Times New Roman","serif""><b><span style="font-size:10.0pt"><span style="line-height:150%"><span style="font-family:"Arial","sans-serif"">Results:</span></span></span></b><b> </b><span style="font-size:10.0pt"><span style="line-height:150%">In pluripotent stem cells and germ cells, the expression levels of </span></span><i><span style="font-size:10.0pt"><span style="line-height:150%">Gfra1</span></span></i><span style="font-size:10.0pt"><span style="line-height:150%">, </span></span><i><span style="font-size:10.0pt"><span style="line-height:150%">Lin28</span></span></i><span style="font-size:10.0pt"><span style="line-height:150%">, and </span></span><i><span style="font-size:10.0pt"><span style="line-height:150%">Sall4</span></span></i><span style="font-size:10.0pt"><span style="line-height:150%"> genes were found </span></span><span dir="RTL" lang="FA" style="font-size:10.0pt"><span style="line-height:150%"><span style="font-family:"Cambria","serif"">t</span></span></span><span style="font-size:10.0pt"><span style="line-height:150%">o</span></span><i> </i><span style="font-size:10.0pt"><span style="line-height:150%">be significantly positive (P<0.05). Alkaline phosphatase was also expressed positively in germ stem cells and pluripotent stem cells, but not in Sertoli cells or fibroblasts, indicating that this enzyme is exclusive to these cell types.</span></span></span></span></span><br>
<span style="font-size:12pt"><span style="line-height:150%"><span style="vertical-align:top"><span style="font-family:"Times New Roman","serif""><b><span style="font-size:10.0pt"><span style="line-height:150%"><span style="font-family:"Arial","sans-serif"">Conclusion:</span></span></span></b> <span style="font-size:10.0pt"><span style="line-height:150%">Because of their interaction and expression in pluripotent stem cells and germ cells, alkaline phosphatase, </span></span><i><span style="font-size:10.0pt"><span style="line-height:150%">Gfra1</span></span></i><span style="font-size:10.0pt"><span style="line-height:150%">, </span></span><i><span style="font-size:10.0pt"><span style="line-height:150%">Lin28</span></span></i><span style="font-size:10.0pt"><span style="line-height:150%">, and </span></span><i><span style="font-size:10.0pt"><span style="line-height:150%">Sall4</span></span></i><span style="font-size:10.0pt"><span style="line-height:150%"> genes can be employed as detecting markers for these cell lines.</span></span><i><span style="font-size:10.0pt"><span style="line-height:150%"></span></span></i></span></span></span></span><br>
سلول های بنیادی جنینی, سلول های بنیادی اسپرم ساز (SSC), آلکالین فسفاتاز
embryonic stem cells, spermatogonial stem cells, alkaline phosphatase
13
25
http://jmums.mazums.ac.ir/browse.php?a_code=A-10-15484-1&slc_lang=fa&sid=1
Emad
Reza
عماد
رضا
e.reza@ausmt.ac.ir
1003194753284600145816
1003194753284600145816
No
BSc in Cellular and Molecular Biology, Department of Biotechnology, Faculty of Biotechnology, Amol University of Special Modern Technologies, Amol, Iran
کارشناسی زیست شناسی سلولی و مولکولی، دانشکده زیست فناوری، دانشگاه تخصصی فناوری های نوین آمل، آمل، ایران
Hossein
Azizi
حسین
عزیزی
h.azizi@ausmt.ac.ir
1003194753284600145817
1003194753284600145817
Yes
Assistant Professor, Department of Biotechnology, Faculty of Biotechnology, Amol University of Special Modern Technologies, Amol, Iran
استادیار، گروه زیست فناوری، دانشکده زیست فناوری، دانشگاه تخصصی فناوری های نوین آمل، آمل، ایران