RESEARCH OF DIFFERENT EXPRESSIONS OF microRNA IN SINUS NODE, ATRIAL MYOCARDIUM, AND VENTRICULAR MYOCARDIUM OF MOUSE_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

FANGYibing ¹ , WENZhi ¹ , DENGMingbin ¹ , LAIQiancheng ² , YUFengxu ¹ , TANGXiaojun ¹ , WUChangxue ¹ , MUGuangrong ¹ ,  LIAOBin ¹

1. Department of Cardiothoracic Surgery, the Affiliated Hospital, Luzhou Medical College, Luzhou Sichuan, 646000, P. R. China;
2. Department of Cardiothoracic Surgery, Chengdu Second People's Hospital;

Corresponding author：

LIAOBin, Email: liaobin@tom.com

Keywords：

microRNA; Differentially expressed gene; Sinus node; Atrial myocardium; Ventricular myocardium; Mouse

DOI：

10.7507/1002-1892.20140247

Video：

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Objective To study microRNA (miRNA) involved in the regulation of sinus cell differentiation by comparing sinus node, atrial myocardium, and ventricular myocardium specific miRNA expression profile differences in Kunming mice. Methods A total of 180 Kunming mice, aged 60-90 days and weighing 35-45 g, were selected without gender differences after the method of anatomical localization for sinus node had been confirmed by preliminary experiments in another 10 Kunming mice. All the sinus node, atrial myocardium, and ventricular myocardium tissue from 180 mice were dissected and frozen by liquid nitrogen. The structure of tissue was observed by HE staining. Total RNA were extracted and quality-controlled before hybridize with miRNA chip. The chips with miRNA were used to screen specific miRNAs; and correlation analysis of gene function was done. Results The area of mice sinus node located at juncture of the superior vena cava and the right atrium junction with crista as its longitudinal axis, ranged 2.0 mm×1.5 mm×1.0 mm. HE staining showed the sinus cells were less, with no stripes, lightly stained cytoplasm, large and round nucleus, and there were much fibrous connective tissue around cells with a visible sinus node artery. The miRNA microarray results showed that compared with atrial myocardium and ventricular myocardium, there were 39 differentially expressed miRNAs in sinus node, including 12 up-regulated miRNAs and 27 down-regulated miRNAs. Based on the regulatory networks of differential miRNA and target gene, the regulatory miRNA was obtained. Conclusion The differentially expressed miRNA in mice sinus node possibly may be involved in the regulation of sinus cell differentiation.

Citation： FANGYibing, WENZhi, DENGMingbin, LAIQiancheng, YUFengxu, TANGXiaojun, WUChangxue, MUGuangrong, LIAOBin. RESEARCH OF DIFFERENT EXPRESSIONS OF microRNA IN SINUS NODE, ATRIAL MYOCARDIUM, AND VENTRICULAR MYOCARDIUM OF MOUSE. Chinese Journal of Reparative and Reconstructive Surgery, 2014, 28(9): 1139-1144. doi: 10.7507/1002-1892.20140247 Copy

1.	Bagnall RD, Tsoutsman T, Shephard RE, et al.Global microRNA profiling of the mouse ventricles during development of severe hypertrophic cardiomyopathy and heart failure.PLoS One, 2012, 7(9):e44744.
2.	Jamshed N, Dubin J, Eldadah Z.Emergency management of palpitations in the elderly:epidemiology, diagnostic approaches, and therapeutic options.Clin Geriatr Med, 2013, 29(1):205-230.
3.	Tseng GN.Can biologic pacemakers respond to physiologic emotional arousal? Heart Rhythm, 2010, 7(12):1841-1842.
4.	Small EM, Olson EN.Pervasive roles of microRNAs in cardiovascular biology.Nature, 2011, 469(7330):336-342.
5.	Espinoza-Lewis RA, Wang DZ.MicroRNAs in heart development.Curr Top Dev Biol, 2012, 100:279-317.
6.	Marvin WJ Jr, Chittick VL, Rosenthal JK, et al.The isolated sinoatrial node cell in primary culture from the newborn rat.Circ Res, 1984, 55(2):253-260.
7.	Lagos-Quintana M, Rauhut R, Lendeckel W, et al.Identification of novel genes coding for small expressed RNAs.Science, 2001, 294(5543):853-858.
8.	Keith A, Flack M.The form and nature of the muscular connections between the primary divisions of the vertebrate heart.J Anat Physiol, 1907, 41(Pt 3):172-189.
9.	魏益平, 朱进国, 张惠忠, 等.原代培养乳鼠窦房结细胞的形态和kir2.1蛋白表达.南方医科大学学报, 2007, 27(11):1701-1705.
10.	Lee I, Ajay SS, Chen H, et al.Discriminating single-base difference miRNA expressions using microarray Probe Design Guru (ProDeG).Nucleic Acids Res, 2008, 36(5):e27.
11.	Jensen SG, Lamy P, Rasmussen MH, et al.Evaluation of two commercial global miRNA expression profiling platforms for detection of less abundant miRNAs.BMC Genomics, 2011, 12:435.
12.	Felli N, Fontana L, Pelosi E, et al.MicroRNAs 221 and 222 inhibit normal erythropoiesis and erythroleukemic cell growth via kit receptor down-modulation.Proc Natl Acad Sci U S A, 2005, 102(50):18081-18086.
13.	Tay YM, Tam WL, Ang YS, et al.MicroRNA-134 modulates the differentiation of mouse embryonic stem cells, where it causes post-transcriptional attenuation of Nanog and LRH1.Stem Cells, 2008, 26(1):17-29.
14.	刘如秀, 谭双, 李敏, 等.强心复脉颗粒药物血清对模拟缺血再灌注致钙超载窦房结细胞的影响.中国中西医结合杂志, 2008, 28(9):828-831.
15.	钟理, 宋治远.原代培养乳鼠窦房结细胞的形态学研究.第三军医大学学报, 2002, 24(4):431-433.
16.	管思彬, 马爱群, 蒋文慧.原代培养乳鼠窦房结细胞的形态学及表面抗原研究.西安交通大学学报:医学版, 2009, 30(3):288-291, 310.
17.	蒋伟, 法宪恩, 李晓召.乳鼠窦房结细胞的分离纯化与形态学研究.医学信息(中旬刊), 2009, 22(12):1075-1076, 1082.
18.	Severs NJ, Slade AM, Powell T, et al.Correlation of ultrastructure and function in calcium-tolerant myocytes isolated from the adult rat heart.J Ultrastruct Res, 1982, 81(2):222-239.
19.	Lin RC, Weeks KL, Gao XM, et al.PI3K (p110 alpha) protects against myocardial infarction-induced heart failure:identification of PI3K-regulated miRNA and mRNA.Arterioscler Thromb Vasc Biol, 2010, 30(4):724-732.
20.	van Rooij E.The art of microRNA research.Circ Res, 2011, 108(2):219-234.
21.	Sun Y, Koo S, White N, et al.Development of a micro-array to detect human and mouse microRNAs and characterization of expression in human organs.Nucleic Acids Res, 2004, 32(22):e188.
22.	Fleissner F, Jazbutyte V, Fiedler J, et al.Short communication:asymmetric dimethylarginine impairs angiogenic progenitor cell function in patients with coronary artery disease through a microRNA-21-dependent mechanism.Circ Res, 2010, 107(1):138-143.
23.	van Mil A, Grundmann S, Goumans MJ, et al.MicroRNA-214 inhibits angiogenesis by targeting Quaking and reducing angiogenic growth factor release.Cardiovasc Res, 2012, 93(4):655-665.
24.	Liu G, Friggeri A, Yang Y, et al.miR-147, a microRNA that is induced upon Toll-like receptor stimulation, regulates murine macrophage inflammatory responses.Proc Natl Acad Sci U S A, 2009, 106(37):15819-15824.
25.	Huppi K, Pitt J, Wahlberg B, et al.Genomic instability and mouse microRNAs.Toxicol Mech Methods, 2011, 21(4):325-333.
26.	Redshaw N, Camps C, Sharma V, et al.TGF-beta/Smad2/3 signaling directly regulates several miRNAs in mouse ES cells and early embryos.PLoS One, 2013, 8(1):e55186.
27.	Dai N, Zhong ZY, Cun YP, et al.Alteration of the microRNA expression profile in human osteosarcoma cells transfected with APE1 siRNA.Neoplasma, 2013, 60(4):384-394.
28.	Song L, Liu H, Gao S, et al.Cellular microRNAs inhibit replication of the H1N1 influenza A virus in infected cells.J Virol, 2010, 84(17):8849-8860.
29.	Sun Z, Wei Q, Zhang Y, et al.MicroRNA profiling of rhesus macaque embryonic stem cells.BMC Genomics, 2011, 12:276.
30.	Bhattacharyya S, Sul K, Krukovets I, et al.Novel tissue-specific mechanism of regulation of angiogenesis and cancer growth in response to hyperglycemia.J Am Heart Assoc, 2012, 1(6):e5967.
31.	Gimble JM, Morgan C, Kelly K, et al.Bone morphogenetic proteins inhibit adipocyte differentiation by bone marrow stromal cells.J Cell Biochem, 1995, 58(3):393-402.
32.	Chartoumpekis DV, Zaravinos A, Ziros PG, et al.Differential expression of microRNAs in adipose tissue after long-term high-fat diet-induced obesity in mice.PLoS One, 2012, 7(4):e34872.
33.	Tsai NP, Lin YL, Wei LN.MicroRNA mir-346 targets the 5'-untranslated region of receptor-interacting protein 140(RIP140) mRNA and up-regulates its protein expression.Biochem J, 2009, 424(3):411-418.
34.	Gantier MP, Stunden HJ, McCoy CE, et al.A miR-19 regulon that controls NF-kB signaling.Nucleic Acids Res, 2012, 40(16):8048-8058.
35.	Garbacki N, Di Valentin E, Huynh-Thu VA, et al.MicroRNAs profiling in murine models of acute and chronic asthma:a relationship with mRNAs targets.PLoS One, 2011, 6(1):e16509.

1. Bagnall RD, Tsoutsman T, Shephard RE, et al.Global microRNA profiling of the mouse ventricles during development of severe hypertrophic cardiomyopathy and heart failure.PLoS One, 2012, 7(9):e44744.
2. Jamshed N, Dubin J, Eldadah Z.Emergency management of palpitations in the elderly:epidemiology, diagnostic approaches, and therapeutic options.Clin Geriatr Med, 2013, 29(1):205-230.
3. Tseng GN.Can biologic pacemakers respond to physiologic emotional arousal? Heart Rhythm, 2010, 7(12):1841-1842.
4. Small EM, Olson EN.Pervasive roles of microRNAs in cardiovascular biology.Nature, 2011, 469(7330):336-342.
5. Espinoza-Lewis RA, Wang DZ.MicroRNAs in heart development.Curr Top Dev Biol, 2012, 100:279-317.
6. Marvin WJ Jr, Chittick VL, Rosenthal JK, et al.The isolated sinoatrial node cell in primary culture from the newborn rat.Circ Res, 1984, 55(2):253-260.
7. Lagos-Quintana M, Rauhut R, Lendeckel W, et al.Identification of novel genes coding for small expressed RNAs.Science, 2001, 294(5543):853-858.
8. Keith A, Flack M.The form and nature of the muscular connections between the primary divisions of the vertebrate heart.J Anat Physiol, 1907, 41(Pt 3):172-189.
9. 魏益平, 朱进国, 张惠忠, 等.原代培养乳鼠窦房结细胞的形态和kir2.1蛋白表达.南方医科大学学报, 2007, 27(11):1701-1705.
10. Lee I, Ajay SS, Chen H, et al.Discriminating single-base difference miRNA expressions using microarray Probe Design Guru (ProDeG).Nucleic Acids Res, 2008, 36(5):e27.
11. Jensen SG, Lamy P, Rasmussen MH, et al.Evaluation of two commercial global miRNA expression profiling platforms for detection of less abundant miRNAs.BMC Genomics, 2011, 12:435.
12. Felli N, Fontana L, Pelosi E, et al.MicroRNAs 221 and 222 inhibit normal erythropoiesis and erythroleukemic cell growth via kit receptor down-modulation.Proc Natl Acad Sci U S A, 2005, 102(50):18081-18086.
13. Tay YM, Tam WL, Ang YS, et al.MicroRNA-134 modulates the differentiation of mouse embryonic stem cells, where it causes post-transcriptional attenuation of Nanog and LRH1.Stem Cells, 2008, 26(1):17-29.
14. 刘如秀, 谭双, 李敏, 等.强心复脉颗粒药物血清对模拟缺血再灌注致钙超载窦房结细胞的影响.中国中西医结合杂志, 2008, 28(9):828-831.
15. 钟理, 宋治远.原代培养乳鼠窦房结细胞的形态学研究.第三军医大学学报, 2002, 24(4):431-433.
16. 管思彬, 马爱群, 蒋文慧.原代培养乳鼠窦房结细胞的形态学及表面抗原研究.西安交通大学学报:医学版, 2009, 30(3):288-291, 310.
17. 蒋伟, 法宪恩, 李晓召.乳鼠窦房结细胞的分离纯化与形态学研究.医学信息(中旬刊), 2009, 22(12):1075-1076, 1082.
18. Severs NJ, Slade AM, Powell T, et al.Correlation of ultrastructure and function in calcium-tolerant myocytes isolated from the adult rat heart.J Ultrastruct Res, 1982, 81(2):222-239.
19. Lin RC, Weeks KL, Gao XM, et al.PI3K (p110 alpha) protects against myocardial infarction-induced heart failure:identification of PI3K-regulated miRNA and mRNA.Arterioscler Thromb Vasc Biol, 2010, 30(4):724-732.
20. van Rooij E.The art of microRNA research.Circ Res, 2011, 108(2):219-234.
21. Sun Y, Koo S, White N, et al.Development of a micro-array to detect human and mouse microRNAs and characterization of expression in human organs.Nucleic Acids Res, 2004, 32(22):e188.
22. Fleissner F, Jazbutyte V, Fiedler J, et al.Short communication:asymmetric dimethylarginine impairs angiogenic progenitor cell function in patients with coronary artery disease through a microRNA-21-dependent mechanism.Circ Res, 2010, 107(1):138-143.
23. van Mil A, Grundmann S, Goumans MJ, et al.MicroRNA-214 inhibits angiogenesis by targeting Quaking and reducing angiogenic growth factor release.Cardiovasc Res, 2012, 93(4):655-665.
24. Liu G, Friggeri A, Yang Y, et al.miR-147, a microRNA that is induced upon Toll-like receptor stimulation, regulates murine macrophage inflammatory responses.Proc Natl Acad Sci U S A, 2009, 106(37):15819-15824.
25. Huppi K, Pitt J, Wahlberg B, et al.Genomic instability and mouse microRNAs.Toxicol Mech Methods, 2011, 21(4):325-333.
26. Redshaw N, Camps C, Sharma V, et al.TGF-beta/Smad2/3 signaling directly regulates several miRNAs in mouse ES cells and early embryos.PLoS One, 2013, 8(1):e55186.
27. Dai N, Zhong ZY, Cun YP, et al.Alteration of the microRNA expression profile in human osteosarcoma cells transfected with APE1 siRNA.Neoplasma, 2013, 60(4):384-394.
28. Song L, Liu H, Gao S, et al.Cellular microRNAs inhibit replication of the H1N1 influenza A virus in infected cells.J Virol, 2010, 84(17):8849-8860.
29. Sun Z, Wei Q, Zhang Y, et al.MicroRNA profiling of rhesus macaque embryonic stem cells.BMC Genomics, 2011, 12:276.
30. Bhattacharyya S, Sul K, Krukovets I, et al.Novel tissue-specific mechanism of regulation of angiogenesis and cancer growth in response to hyperglycemia.J Am Heart Assoc, 2012, 1(6):e5967.
31. Gimble JM, Morgan C, Kelly K, et al.Bone morphogenetic proteins inhibit adipocyte differentiation by bone marrow stromal cells.J Cell Biochem, 1995, 58(3):393-402.
32. Chartoumpekis DV, Zaravinos A, Ziros PG, et al.Differential expression of microRNAs in adipose tissue after long-term high-fat diet-induced obesity in mice.PLoS One, 2012, 7(4):e34872.
33. Tsai NP, Lin YL, Wei LN.MicroRNA mir-346 targets the 5'-untranslated region of receptor-interacting protein 140(RIP140) mRNA and up-regulates its protein expression.Biochem J, 2009, 424(3):411-418.
34. Gantier MP, Stunden HJ, McCoy CE, et al.A miR-19 regulon that controls NF-kB signaling.Nucleic Acids Res, 2012, 40(16):8048-8058.
35. Garbacki N, Di Valentin E, Huynh-Thu VA, et al.MicroRNAs profiling in murine models of acute and chronic asthma:a relationship with mRNAs targets.PLoS One, 2011, 6(1):e16509.

Chinese Journal of Reparative and Reconstructive Surgery

RESEARCH OF DIFFERENT EXPRESSIONS OF microRNA IN SINUS NODE, ATRIAL MYOCARDIUM, AND VENTRICULAR MYOCARDIUM OF MOUSE

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