الگوی بیانی ژن p32 در بافت‌های ران، دست، راسته و چربی پشت بره کرمانی

نوع مقاله : مقاله پژوهشی

نویسندگان

1 بخش علوم دامی، دانشگاه شهید باهنر کرمان

2 بخش علوم دامی، دانشکده کشاورزی، دانشگاه شهید باهنر کرمان

3 دانشیار، گروه علوم دامی، دانشکده کشاورزی، دانشگاه شهید باهنر کرمان، کرمان، ایران

چکیده

هدف: مطالعات نشان داده‌اند که p32 در بافت‌های فعال متابولیکی و به سرعت در حال رشد، مانند ماهیچه‌ها، تومورهای سینه، اپیدرم و تخمدان به شدت بیان می‌شود. یکی از اقدامات اساسی در حیوانات اهلی مطالعه ژن­ها و پروتئین­های مرتبط با صفات اقتصادی و مطالعه آنها در سطح سلولی یا کروموزومی است. هدف این مطالعه، بررسی الگوی بیانی ژن p32 در بافت‌های ران، دست، راسته و چربی پشت کمر بره کرمانی بود.
مواد و روش‌ها: نمونه‌برداری از بافت‌های ران، دست، راسته و چربی پشت کمر بره‌های نر کرمانی (12 نمونه از 3 حیوان و از هر بافت 3 تکرار) انجام شد. پس از انجماد سریع نمونه ها در داخل فریزر 80- نگهداری شدند. RNA کل با استفاده از کیت استاندارد استخراج شد. کیفیت و کمیتRNA  استخراج شده با روش الکتروفورز روی ژل آگارز و با استفاده از دستگاه نانودراپ مورد ارزیابی قرار گرفت. برای سنتز  cDNAاز کیت سنتز cDNA شرکت parstous استفاده شد. برای بررسی میزان نسبی بیان ژن‌ها از واکنش real time PCR به روش Syber Green استفاده شد. برای تجزیه و تحلیل داده های حاصل از real time PCR از روش Pfaffl et al. استفاده شد.
نتایج: بررسی کیفیت RNA­های استخراج شده نشان داد که کیفیت مناسب و مطلوب است. مشاهده دو باند 18S و 28S در RNA نشان دهنده سالم بودن آن و عدم وجود باند اضافی نشان دهنده خلوص آن بود. الکتروفورز محصولات PCR روی ژل آگارز و نتایج منحنی­های real time PCR نشان داد که ژن p32 در بافت­های ران، راسته و چربی پشت کمر بره‌های نر کرمانی بیان نشده است و فقط در بافت دست این حیوانات تکثیر شده است. مشاهده تک باند در محدوده bp281 برای ژن p32 در بافت­ دست و وجود باند در محدوده bp112 برای ژن بتااکتین در همه­ نمونه­ها، بیانگر صحت انجام آزمایش و تکثیر قطعه مورد نظر بود.
نتیجه‌گیری: با توجه به این که در این مطالعه بیان ژن p32 در گوسفند کرمانی در سن 11 ماهگی مطالعه شد و بیان این ژن بیشتر در دوران جنینی گزارش شده است، لذا عدم مشاهده بیان این ژن در بافت‌های ران، راسته و چربی پشت و بیان پایین آن در بافت دست تایید کننده نتایج مطالعات قبلی است. بیان کم این ژن در عضله دست می‌تواند حاکی از این باشد که این عضله هنوز در این سن دارای رشد است و یا ممکن است این بافت سرطانی شده باشد، چرا که این ژن بیشتر در بافت‌هایی بیان می‌شود که در حال رشد سریع هستند و یا درگیر سرطان هستند.

کلیدواژه‌ها


عنوان مقاله [English]

The expression pattern of p32 gene in femur, humeral muscle, back muscle and back fat tissues of Kermani lambs

نویسندگان [English]

  • zahra arabpour 1
  • Mohammadreza Mohammadabadi 2
  • Amin Khezri 3
1 : MSc Student, Animal Science Department, Faculty of Agriculture, Shahid Bahonar University of Kerman, Kerman, Iran
2 Department of Animal Science, Faculty of Agriculture, College of Agriculture, Shahid Bahonar University of Kerman
3 Associate Professor, Department of Animal Science, College of Agriculture, Shahid Bahonar University of Kerman, Kerman, Iran.
چکیده [English]

Objective
Studies have shown that p32 is highly expressed in metabolically active and rapidly growing tissues, such as muscles, breast tumors, epidermis, and ovaries. One of the basic measures in domestic animals is the study of genes and proteins related to economic traits and their study at the cellular or chromosomal level. The aim of this study was to investigate the expression pattern of p32 gene in femur, humeral muscle, back muscle and back fat tissues of Kermani lambs.
 
Materials and Methods
Samples of femur, humeral muscle, back muscle and back fat tissues of Kermani lambs were collected (12 samples from 3 animals and 3 replicates from each tissue). After rapid freezing, the samples were stored in a -80 freezer. Total RNA was extracted using a standard kit. The quality and quantity of the extracted RNA were evaluated by agarose gel electrophoresis and using a nanodrop device. For cDNA synthesis, parstous cDNA synthesis kit was used. Real time PCR reaction by Syber Green method was used to evaluate the relative expression of genes. To analyze the data obtained from real time PCR, the Pfaffl method was used.
 
 
Results
The quality of the extracted RNAs showed that the quality is appropriate and desirable. Observation of 18S and 28S bands in RNA indicated that it was intact and that no additional bands indicated its purity. Electrophoresis of PCR products on agarose gel and the results of real time PCR curves showed that the p32 gene was not expressed in femur, back muscle and back fat tissues of Kermani male lambs and was amplified only in the humeral muscle tissue of these animals. The observation of a single band in the range of 281 bp for the p32 gene in the humeral muscle tissue and the presence of a band in the range of 112 bp for the beta actin gene in all samples indicated the accuracy of the experiment and the amplification of the fragment.
 
Conclusion
Considering that in this study the expression of p32 gene in Kermani sheep at the age of 11 months was studied and the expression of this gene has been reported more in the embryonic period, so no expression of this gene was observed in femur, back muscle and back fat and its low expression in the humeral muscle tissue confirms the results of previous studies. Low expression of this gene in the humeral muscle tissue may indicate that the muscle is still growing at this age or that the tissue may be cancerous, as this gene is more expressed in tissues that are growing rapidly or are involved in cancer.

کلیدواژه‌ها [English]

  • Expression pattern
  • p32 gene
  • tissue
  • Kermani sheep
توحیدی نژاد فاطمه، محمدآبادی محمدرضا، اسمعیلی زاده کشکوئیه علی، نجمی نوری عذرا (1393). مقایسه سطوح مختلف بیان ژنRheb  در بافت های مختلف بز کرکی راینی. مجله بیوتکنولوژی کشاورزی 6(4)، 50-35.
جعفری دره در امیر حسین، محمدآبادی محمدرضا، اسمعیلی زاده کشکوئیه علی، ریاحی مدوار علی (1395) بررسی بیان ژن CIB4  در بافت­های مختلف گوسفند کرمانی با استفاده از real time qPCR. مجله پژوهش در نشخوارکنندگان 4(4)، 132-119.
محمدآبادی محمدرضا (1399) بیان ژن ESR1 در بز کرکی راینی با استفاده از real time PCR‎. مجله بیوتکنولوژی کشاورزی 12(1)، 192-177.
محمدآبادی محمدرضا، سفلایی محمد (1399). پروفایل بیانی mRNA مختص بافت ژن BMP15 در بز. مجله بیوتکنولوژی کشاورزی 12(3)، 208-191.
محمدآبادی محمدرضا، کرد محبوبه، نظری محمود (1397) مطالعه بیان ژن لپتین در بافت‌های مختلف گوسفند کرمانی با استفاده از real time PCR. مجله بیوتکنولوژی کشاورزی 10(3)، 122-111.
محمدی فر آمنه، محمدآبادی محمدرضا (1390) کاربرد نشانگرهای ریزماهواره برای مطالعه ژنوم گوسفند کرمانی. مجله علوم دامی ایران 42(4)، 344-337.
واجدابراهیمی محمدتقی، محمدآبادی محمدرضا، اسماعیلی زاده علی (1394) بررسی تنوع ژنتیکی پنج جمعیت گوسفند ایرانی با استفاده از نشانگرهای ریزماهواره­ای. مجله بیوتکنولوژی کشاورزی 7(4)، 158-143.
 
References
Amiri Roudbar M, Mohammadabadi MR, Mehrgardi AA, Abdollahi-Arpanahi A (2017) Estimates of variance components due to parent-of-origin effects for body weight in Iran-Black sheep. Small Rum Res 149, 1-5
Amiri Roudbar M., Abdollahi-Arpanahi R., Ayatollahi Mehrgardi A., et al. (2018) Estimation of the variance due to parent-of-origin effects for productive and reproductive traits in Lori-Bakhtiari sheep. Small Rumin Res 160, 95-102.
Byrne K, Vuocolo T, Gondro C, et al. (2010) A gene network switch enhances the oxidative capacity of ovine skeletal muscle during late fetal development. BMC Genomics 11, e378.
Chargé SBP, Rudnicki MA (2004) Cellular and molecular regulation of muscle regeneration. Physiol Rev 84, 209–238.
Chen YB, Jiang CT, Zhang GQ, et al. (2010) Increased expression of hyaluronic acid binding protein 1 is correlated with poor prognosis in patients with breast cancer. J Surg Oncol 100, 382–386.
Dedio J, Jahnen-Dechent W, Bachmann M, Müller-Esterl W (1998) The multiligand-binding protein gC1qR, putative C1q receptor, is a mitochondrial protein. J Immunol 160, 3534.
Edmondson DG, Lyons GE, Martin JF, Olson EN (1994) Mef2 gene expression marks the cardiac and skeletal muscle lineages during mouse embryogenesis. Development 120, 1251–1263.
Fahey AJ, Brameld JM, Parr T, rt al. (2005) The effect of maternal undernutrition before muscle differentiation on the muscle fiber development of the newborn lamb. J Anim Sci 83, e2564.
Fogal V, Richardson AD, Karmali PP, et al. (2010) Mitochondrial p32 Protein is a Critical Regulator of Tumor Metabolism via Maintenance of Oxidative Phosphorylation. Mol Cell Biol 30, e1303.
Ghotbaldini H., Mohammadabadi M.R., Nezamabadi-pour H., Babenko O.I., Bushtruk M.V. and Tkachenko S.V. (2019). Predicting breeding value of body weight at 6-month age using Artificial Neural Networks in Kermani sheep breed. Acta Scientiarum. Anim Sci 41, e45282.
Gwinn DM, Shackelford DB, Egan DF, et al. (2008) AMPK Phosphorylation of Raptor Mediates a Metabolic Checkpoint. Mol Cell 30, 214–226.
Hardie DG (2007) AMP-activated/SNF1 protein kinases: Conserved guardians of cellular energy. Nat Rev Mol Cell Biol 8, 774–785.
Hemmings KM, Daniel ZCTR, Buttery PJ, et al. (2015) Differential effects of short-term agonist and growth hormone treatments on expression of myosin heavy chain IIB and associated metabolic genes in sheep muscle. Animal 9, 285–294.
Jafari Darehdor AH, Mohammadabadi MR, Esmailizadeh AK, Riahi Madvar A (2016) Investigating expression of CIB4 gene in different tissues of Kermani Sheep using real time qPCR. J Rumin Res 4, 119-132 (In Persian).
Liu Y, Leslie PL, Jin A, et al. (2017) p32 heterozygosity protects against age- and diet-induced obesity by increasing energy expenditure. Sci Rep 7, e5754.
Ma J, Ren C, Yang H, Zhao J, et al. (2019) The Expression Pattern of p32 in Sheep Muscle and Its Role in Differentiation, Cell Proliferation, and Apoptosis of Myoblasts. Int J Mol Sci 20(20), e5161.
Masoudzadeh SH, Mohammadabadi M, Khezri A, et al. (2020a) Effects of diets with different levels of fennel (Foeniculum vulgare) seed powder on DLK1 gene expression in brain, adipose tissue, femur muscle and rumen of Kermani lambs. Small Rumin Res 193, e106276.
Masoudzadeh SH, Mohammadabadi MR, Khezri A, et al. (2020b) Dlk1 Gene Expression in Different Tissues of Lamb. Iran J Appl Anim Sci 10 (4), 669-677.
Matos P, Horn JA, Beards F, et al. (2014) A role for the mitochondrial-associated protein p32 in regulation of trophoblast proliferation. Mol Hum Reprod 20, 745.
Mengjie H, Crawford SA, Henstridge DC (2013) p32 protein levels are integral to mitochondrial and endoplasmic reticulum morphology, cell metabolism and survival. Biochem J 453, 381–391.
Mikako Y, Takeshi U, Shinya T, et al. (2012) p32/gC1qR is indispensable for fetal development and mitochondrial translation: Importance of its RNA-binding ability. Nucleic Acids Res 40, 9717–9737.
Mohammadabadi M, Masoudzadeh SH, Khezri A, et al. (2021) Fennel (Foeniculum vulgare) seed powder increases Delta-Like Non-Canonical Notch Ligand 1 gene expression in testis, liver, and humeral muscle tissues of growing lambs. Heliyon 7 (12), e08542.
Mohammadabadi M, Soflaei M (2020) Tissue-specific mRNA expression profile of BMP15 gene in goat. Agric Biotechnol J 12, 191-208 (In Persian).
Mohammadabadi MR (2016) Role of clostridium perfringens in pathogenicity of some domestic animals. J Adv Agric 7 (3), 1117-1121.
Mohammadabadi MR (2017) Inter-Simple Sequence Repeat loci Associations with Predicted Breeding Values of Body Weight in Kermani Sheep. Genet 3rd Millen 14 (4), 4383-4390.
Mohammadabadi MR (2020) Expression of ESR1 gene in Raini Cashmere goat using real time PCR. Agric Biotechnol J 12 (1), 177-192 (In Persian).
Mohammadabadi MR, Esfandyarpoor E, Mousapour A (2017) Using Inter Simple Sequence Repeat Multi-Loci Markers for Studying Genetic Diversity in Kermani Sheep. J Res Dev 5 (2), e154.
Mohammadabadi MR, Kord M, Nazari M (2018) Studying expression of leptin gene in different tissues of Kermani Sheep using real time PCR. Agric Biotechnol J 10, 111-122 (in Persian).
Mohammadifar A, Mohammadabadi MR (2011) Application of Microsatellite Markers for a Study of Kermani Sheep Genome. Iran J Anim Sci 42 (4), 337-344 (In Persian).
Muta T, Kang D, Kitajima S, et al. (1997) p32 protein, a splicing factor 2-associated protein, is localized in mitochondrial matrix and is functionally important in maintaining oxidative phosphorylation. J Biol Chem 272, 24363.
Naya FJ (1999) MEF 2: A transcriptional target for signaling pathways controlling skeletal muscle growth and di_erentiation. Curr. Opin. Cell Biol 11, 683–688.
Nguyen N, Kim KB, Yi JS (2011) Cell-surface receptor for complement component Clq(gClqR) is a key regulator for lamellipodia formation and cancer metastasis. J Biol Chem 286, 23093–23101.
Parker MH, Patrick S, Rudnicki MA (2003) Looking back to the embryo: Defining transcriptional networks in adult myogenesis. Nat Rev Genet 4, 497–507.
Pasandideh M, Kharrati Koopaee H, Mohammad Abadi MR, et al. (2016) Association of the OPN and PPARGC1A Alleles with Milk Somatic Cell Count in Iranian Holstein cattle. Modern Genet J 11, 357-365 (In Persian).
Peterson KL, Zhang W, Lu PD, et al. (1997) The C1q-binding cell membrane proteins cC1q-R and gC1q-R are released from activated cells: Subcellular distribution and immunochemical characterization. Clin Immunol Immunopathol 84, 17–26.
Pfaffl MW, Horgan GW, Dempfle L (2002) Relative expression software tool (REST©) for group-wise comparison and statistical analysis of relative expression results in real-time PCR. Nucleic Acids Res 30, e36.
Ren C, Deng M, Fan Y, et al. (2017) Genome-Wide Analysis Reveals Extensive Changes in LncRNAs during Skeletal Muscle Development in Hu Sheep. Genes 8, 283.
Ren HX, Li L, Su HW, et al. (2011) Histological and Transcriptome-wide Level Characteristics of Fetal Myofiber Hyperplasia During the Second Half of Gestation in Texel and Ujumqin Sheep. BMC Genom 12, 411.
Rion N, Castets P, Lin S, et al. (2019) mTOR controls embryonic and adult myogenesis via mTORC1. Development 146, dev172460.
Saito T, Uchiumi T, Yagi M, et al. (2017) Cardiomyocyte-specific loss of mitochondrial p32/C1qbp causes cardiomyopathy and activates stress responses. Cardiovasc Res 113, 1173–1185.
Shahsavari M, Mohammadabadi M, Khezri A, et al. (2021) Correlation between insulin-like growth factor 1 gene expression and fennel (Foeniculum vulgare) seed powder consumption in muscle of sheep. Animal Biotechnology 33, 1-11.
Shaw RJ, Monica K, Nabeel B, et al. (2004) The tumor suppressor LKB1 kinase directly activates AMP-activated kinase and regulates apoptosis in response to energy stress. Proc Natl Acad Sci USA 101, 3329–3335.
Song X, Kim SY, Zhang L, et al. (2014) Role of AMP-activated protein kinase in cross-talk between apoptosis and autophagy in human colon cancer. Cell Death Dis 5, e1504.
Tohidi nezhad F, Mohammadabadi MR, Esmailizadeh AK, Najmi Noori A (2015) Comparison of different levels of Rheb gene expression in different tissues of Raini Cashmir goat. Agric Biotechnol J 6, 35-50 (In Persian).
Vajed Ebrahimi MT, Mohammad Abadi MR, Esmailizadeh AK (2016) Analysis of genetic diversity in five Iranian sheep population using microsatellites markers. Agric Biotechnol J 7 (4), 143-158 (In Persian).
Valentina F, Ivan B, Ying C, et al. (2015) Mitochondrial p32 is upregulated in Myc expressing brain cancers and mediates glutamine addiction. Oncotarget 6, 1157–1170.
Valentina F, Lianglin Z, Stan K, Erkki R (2008) Mitochondrial/cell-surface protein p32/gC1qR as a molecular target in tumor cells and tumor stroma. Cancer Res 68, 7210.
Xiaodong F, Tonnesen MG, Peerschke EIB, Berhane G (2002) Cooperation of C1q receptors and integrins in C1q-mediated endothelial cell adhesion and spreading. J Immunol 168, 2441–2448.
Xu Y, Qian H, Feng X, et al. (2012) Differential proteome and transcriptome analysis of porcine skeletal muscle during development. J Proteom 75, 2093–2108.
Yu H, Liu Q, Xin T, et al. (2013) Elevated expression of hyaluronic acid binding protein 1 (HABP1)/P32/C1QBP is a novel indicator for lymph node and peritoneal metastasis of epithelial ovarian cancer patients. Tumor Biol 34, 3981–3987.
Zhang X, Zhang F, Guo L, et al. (2013) Interactome analysis reveals that C1QBP (complement component 1, q subcomponent binding protein) is associated with cancer cell chemotaxis and metastasis. Mol Cell Proteom 12, 3199–3209.
Zhao X, Mo D, Li A, et al. (2011) Comparative Analyses by Sequencing of Transcriptomes during Skeletal Muscle Development between Pig Breeds Differing in Muscle Growth Rate and Fatness. PLoS ONE 6, e19774.