Research Article
BibTex RIS Cite

Kardiyomiyoblast Hücre Regülasyonunda Taube Nuss Geninin Fonksiyonel Analizi

Year 2019, Volume: 2 Issue: 3, 98 - 104, 30.12.2019
https://doi.org/10.26650/JARHS2019-651072

Abstract

Gelişim evrelerinin veya dokuya özgü fizyolojik süreçlerin düzenlenmesinde ve yürütülmesinde farklılaşmış gen ekspresyonu büyük önem taşımaktadır. “Subtractive” hibridizasyon yöntemi dokuya özgü ekspresyonu olan genlerin tespit edilmesi için oldukça etkin bir yöntemdir. Daha önceki çalışmalarda laboratuvarımızda BALB/c ırkı farelerde kalbe özgü ekspresyonu olan genleri içeren “Subtractive” hibridizasyon cDNA kütüphanesi oluşturulmuştur. Önemli bir transkripsiyon faktörü olan Taube Nuss (Tbn), bu kütüphaneden elde edilen genlerden birisidir. Bu çalışmadaki amacımız, Tbn geni ile ilişkili olduğu düşünülen diğer genlerin ekspresyonlarının araştırılması ve Tbn yolağının belirlenmesidir. Çalışma kapsamında BALB/c ırkı farelerden elde edilen kalp ve iskelet kası dokularına özgü Tbn geninin ekspresyonu Northern Blot analizi ile araştırılmıştır. Ardından, Tbn genine özgü siRNA’lar kullanılarak H9c2 rat kardiyomiyoblast hücre soyunda Tbn geni sessizleştirilmiştir. Yolak analizleri sonucunda Tbn ile ilişkili olabileceği tespit edilen Pparg, Cebpa, Taf10 ve Myocd genlerinin sessizleştirilen hücrelerdeki ekspresyon değişimleri araştırılmıştır. Bu hücrelerde, Myocd gen ekspresyonunun azaldığı ve Pparg gen ekspresyonunun arttığı saptanmıştır. Bu sonuçlar, Tbn geninin kalp dokusunda düzenleyici bir rol oynadığını işaret etmektedir. 

Supporting Institution

İstanbul Üniversitesi Bilimsel Araştırma Projeleri Birimi

Project Number

2140

References

  • 1. Calore M., De Windt L.J., Rampazzo A. (2015): Genetics meets epigenetics: genetic variants that modulate noncoding RNA in cardiovascular diseases, J Mol Cell Cardiol, 89: 27-34.
  • 2. Elia L., Condorelli G. (2015): RNA (Epi)genetics in cardiovascular diseases, J Mol Cell Cardiol, 89: 11-16.
  • 3. Diatchenko L., Lau Y.F., Campbell A.P., Chenchik A., Moqadam F., Huang B., Lukyanov S., Lukyanov K., Gurskaya N., Sverdlov E.D., Siebert P.D. (1996): Suppression subtractive hybridization: a method for generating differentially regulated or tissue-specific cDNA probes and libraries, Proc Natl Acad Sci U S A, 93(12): 6025-30.
  • 4. Özsait B. (2003): “Subtractive” Hibridizasyon Kütüphanesinden izole edilen ve kalp gelişiminde rolü olduğu düşünülen genlerin analizi, Yayınlanmamış Yüksek Lisans Tezi, İstanbul Üniversitesi Sağlık Bilimleri Enstitüsü, İstanbul.
  • 5. Komurcu-Bayrak E., Ozsait B., Erginel-Unaltuna N. (2012): Isolation and analysis of genes mainly expressed in adult mouse heart using subtractive hybridization cDNA library, Mol Biol Rep,39(8):8065-74.
  • 6. Ozsait Selcuk B., Kömürcü Bayrak E., Erginal Ünaltuna N. (2016): Higher Expression level of Bat3 is associated with silencing of Midn gene in primary mouse cardiomyocytes, Turk J Biol, 40: 1295-1302.
  • 7. Guermah M., Ge K., Chiang C.M., Roeder R.G. Guermah M., Ge K., Chiang C.M., Roeder R.G. (2003): The TBN protein, which is essential for early embryonic mouse development, is an inducible TAFII implicated in adipogenesis, Mol Cell, 12: 991-1001.
  • 8. Voss A.K., Thomas T., Petrou P., Anastassiadis K., Schöler H., Gruss P. (2000): Taube Nuss is a Novel Gene Essential for the Survival of Pluripotent Cells of Early Mouse Embryos, Development, 127: 5449-61.
  • 9. Brison D.R., Schultz R.M. (1997): Apoptosis during mouse blastocyst formation: evidence for a role for survival factors including transforming growth factor alpha, Biol Reprod, 56: 1088-96.
  • 10. Demény M.A., Soutoglou E., Nagy Z., Scheer E., Jànoshàzi A., Richardot M., Argentini M., Kessler P., Tora L. (2007): Identification of a small TAF complex and its role in the assembly of TAF-containing complexes, PLoS One, 2(3):e316.
  • 11. Trowitzsch S., Viola C., Scheer E., Conic S., Chavant V., Fournier M., Papai G., Ebong I.O., Schaffitzel C., Zou J., Haffke M., Rappsilber J., Robinson C.V., Schultz P., Tora L., Berger I. (2015): Cytoplasmic TAF2-TAF8TAF10 complex provides evidence for nuclear holoTFIID assembly from preformed submodules, Nat Commun, 14(6): 6011.
  • 12. Tatarakis A., Margaritis T., Martinez-Jimenez C.P., Kouskouti A., Mohan W.S. 2nd, Haroniti A., Kafetzopoulos D., Tora L., Talianidis I.. (2008): Dominant and redundant functions of TFIID involved in the regulation of hepatic genes, Mol Cell, 31: 531–543.
  • 13. Mouse ENCODE Consortium. (2014): A comparative encyclopedia of DNA elements in the mouse genome, Nature, 515(7527):355-64.
  • 14. El-Jack A.K., Hamm J.K., Pilch P.F., Farmer S.R.. (1999): Reconstitution of Insulin-sensitive Glucose Transport in Fibroblasts Requires Expression of Both PPARγ and C/EBPα, J Biol Chem, 274: 7946-51.
  • 15. Gordon J.W. (2018): Regulation of cardiac myocyte cell death and differentiation by myocardin, Mol Cell Biochem, 437(1-2): 119-131.
  • 16. Raphel L., Talasila A., Cheung C., Sinha S. (2012): Myocardin overexpression is sufficient for promoting the development of a mature smooth muscle cell-like phenotype from human embryonic stem cells, PLoS One, 7(8): e44052.
  • 17. Singh J., Verma N.K., Kansagra S.M., Kate B.N. Dey C.S. (2007): Altered PPARgamma expression inhibits myogenic differentiation in C2C12 skeletal muscle cells, Molecular and Cellular Biochemistry, 294: 163–171.
  • 18. He K., Wu G., Li W.X., Guan D., Lv W., Gong M., Ye S., Lu A. (2017): A transcriptomic study of myogenic differentiation under the overexpression of PPARγ by RNA-Seq, Sci Rep, 7(1):15308.

Functional Analysis of Taube Nuss Gene in the Regulation of Cardiomyoblast Cells

Year 2019, Volume: 2 Issue: 3, 98 - 104, 30.12.2019
https://doi.org/10.26650/JARHS2019-651072

Abstract

Differential gene expression is important in the regulation and maintenance of developmental stages and tissue specific physiological processes. The subtractive hybridization method is an efficient method for the detection of genes that have tissue specific expressions. In our previous work, our team had constructed a BALB/c mice heart tissue specific subtractive hybridization cDNA library. Taube Nuss (Tbn), which is a crucial transcription factor, is one of the genes that were isolated from this library. In this study, our aim was to analyze the expression of possible Tbn related genes and identify the Tbn pathway. In the scope of this study, the Tbn gene expression in BALB/c mice heart and skeletal tissue was analyzed with the Northern Blot technique. Then, the Tbn gene was silenced in H9c2 rat cardiomyoblast cell line using siRNA specific to the TBN gene. The gene expression levels of Pparg, Cebpa, Taf10 and Myocd genes, which were possibly related to the Tbn pathway, were analyzed in silenced cells. We observed that the expression of the Myocd gene was decreased, whereas the expression of the Pparg gene was increased in these cells. These results suggest that the Tbn gene plays a regulatory role in cardiac tissue. 

Project Number

2140

References

  • 1. Calore M., De Windt L.J., Rampazzo A. (2015): Genetics meets epigenetics: genetic variants that modulate noncoding RNA in cardiovascular diseases, J Mol Cell Cardiol, 89: 27-34.
  • 2. Elia L., Condorelli G. (2015): RNA (Epi)genetics in cardiovascular diseases, J Mol Cell Cardiol, 89: 11-16.
  • 3. Diatchenko L., Lau Y.F., Campbell A.P., Chenchik A., Moqadam F., Huang B., Lukyanov S., Lukyanov K., Gurskaya N., Sverdlov E.D., Siebert P.D. (1996): Suppression subtractive hybridization: a method for generating differentially regulated or tissue-specific cDNA probes and libraries, Proc Natl Acad Sci U S A, 93(12): 6025-30.
  • 4. Özsait B. (2003): “Subtractive” Hibridizasyon Kütüphanesinden izole edilen ve kalp gelişiminde rolü olduğu düşünülen genlerin analizi, Yayınlanmamış Yüksek Lisans Tezi, İstanbul Üniversitesi Sağlık Bilimleri Enstitüsü, İstanbul.
  • 5. Komurcu-Bayrak E., Ozsait B., Erginel-Unaltuna N. (2012): Isolation and analysis of genes mainly expressed in adult mouse heart using subtractive hybridization cDNA library, Mol Biol Rep,39(8):8065-74.
  • 6. Ozsait Selcuk B., Kömürcü Bayrak E., Erginal Ünaltuna N. (2016): Higher Expression level of Bat3 is associated with silencing of Midn gene in primary mouse cardiomyocytes, Turk J Biol, 40: 1295-1302.
  • 7. Guermah M., Ge K., Chiang C.M., Roeder R.G. Guermah M., Ge K., Chiang C.M., Roeder R.G. (2003): The TBN protein, which is essential for early embryonic mouse development, is an inducible TAFII implicated in adipogenesis, Mol Cell, 12: 991-1001.
  • 8. Voss A.K., Thomas T., Petrou P., Anastassiadis K., Schöler H., Gruss P. (2000): Taube Nuss is a Novel Gene Essential for the Survival of Pluripotent Cells of Early Mouse Embryos, Development, 127: 5449-61.
  • 9. Brison D.R., Schultz R.M. (1997): Apoptosis during mouse blastocyst formation: evidence for a role for survival factors including transforming growth factor alpha, Biol Reprod, 56: 1088-96.
  • 10. Demény M.A., Soutoglou E., Nagy Z., Scheer E., Jànoshàzi A., Richardot M., Argentini M., Kessler P., Tora L. (2007): Identification of a small TAF complex and its role in the assembly of TAF-containing complexes, PLoS One, 2(3):e316.
  • 11. Trowitzsch S., Viola C., Scheer E., Conic S., Chavant V., Fournier M., Papai G., Ebong I.O., Schaffitzel C., Zou J., Haffke M., Rappsilber J., Robinson C.V., Schultz P., Tora L., Berger I. (2015): Cytoplasmic TAF2-TAF8TAF10 complex provides evidence for nuclear holoTFIID assembly from preformed submodules, Nat Commun, 14(6): 6011.
  • 12. Tatarakis A., Margaritis T., Martinez-Jimenez C.P., Kouskouti A., Mohan W.S. 2nd, Haroniti A., Kafetzopoulos D., Tora L., Talianidis I.. (2008): Dominant and redundant functions of TFIID involved in the regulation of hepatic genes, Mol Cell, 31: 531–543.
  • 13. Mouse ENCODE Consortium. (2014): A comparative encyclopedia of DNA elements in the mouse genome, Nature, 515(7527):355-64.
  • 14. El-Jack A.K., Hamm J.K., Pilch P.F., Farmer S.R.. (1999): Reconstitution of Insulin-sensitive Glucose Transport in Fibroblasts Requires Expression of Both PPARγ and C/EBPα, J Biol Chem, 274: 7946-51.
  • 15. Gordon J.W. (2018): Regulation of cardiac myocyte cell death and differentiation by myocardin, Mol Cell Biochem, 437(1-2): 119-131.
  • 16. Raphel L., Talasila A., Cheung C., Sinha S. (2012): Myocardin overexpression is sufficient for promoting the development of a mature smooth muscle cell-like phenotype from human embryonic stem cells, PLoS One, 7(8): e44052.
  • 17. Singh J., Verma N.K., Kansagra S.M., Kate B.N. Dey C.S. (2007): Altered PPARgamma expression inhibits myogenic differentiation in C2C12 skeletal muscle cells, Molecular and Cellular Biochemistry, 294: 163–171.
  • 18. He K., Wu G., Li W.X., Guan D., Lv W., Gong M., Ye S., Lu A. (2017): A transcriptomic study of myogenic differentiation under the overexpression of PPARγ by RNA-Seq, Sci Rep, 7(1):15308.
There are 18 citations in total.

Details

Primary Language Turkish
Journal Section Research Articles
Authors

Ayşe Berna Yüzbaşıoğulları This is me 0000-0001-5235-997X

Bilge Özsait Selçuk This is me 0000-0001-6808-6689

Evrim Kömürcü Bayrak 0000-0003-1271-1208

Nihal Erginel Ünaltuna This is me 0000-0003-0562-0455

Project Number 2140
Publication Date December 30, 2019
Submission Date November 26, 2019
Published in Issue Year 2019 Volume: 2 Issue: 3

Cite

MLA Yüzbaşıoğulları, Ayşe Berna et al. “Kardiyomiyoblast Hücre Regülasyonunda Taube Nuss Geninin Fonksiyonel Analizi”. Sağlık Bilimlerinde İleri Araştırmalar Dergisi, vol. 2, no. 3, 2019, pp. 98-104, doi:10.26650/JARHS2019-651072.