The investigation of endoplasmic reticulum stress markers ATF5 and phosphorylated eIF2 after kainic acid treatment in neuroblastoma cells

SARI, Alime; Dönmez Yalçın, Gizem

doi:10.17826/cumj.637075

The investigation of endoplasmic reticulum stress markers ATF5 and phosphorylated eIF2 after kainic acid treatment in neuroblastoma cells

Alime Sarı, (Aydın Adnan Menderes Üniversitesi, Tıp Fakültesi, Tıbbi Biyoloji Anabilim Dalı, Aydın, Türkiye)

Gizem Dönmez Yalçın (Aydın Adnan Menderes Üniversitesi, Tıp Fakültesi, Tıbbi Biyoloji Anabilim Dalı, Aydın, Türkiye)

Cukurova Medical Journal

1 0

Yıl: 2020 Cilt: 45 Sayı: 1 Sayfa Aralığı: 96 - 101 Metin Dili: İngilizce DOI: 10.17826/cumj.637075 İndeks Tarihi: 10-12-2020

The investigation of endoplasmic reticulum stress markers ATF5 and phosphorylated eIF2 after kainic acid treatment in neuroblastoma cells

Öz:

Purpose: The aim of this study was to investigate therelationship between kainic acid induced excitotoxicity andendoplasmic resticulum (ER) stress by analyzing twomajor ER stress markers such as ATF5 andphosphorylated eIF2 in neuroblastoma cells.Materials and Methods: Neuroblastoma cells weretreated with 1 mM kainic acid for 24 hours. ATPmeasurement was performed in kainic acid-treated andvehicle-treated neuroblastoma cells via ATPbioluminescence assay. Total protein was isolated fromkainic acid-treated and control cells. Via western blotting,the expression levels of ATF5 and phosphorylated eIF2were analyzed.Results: We showed for the first time that as a result ofkainic acid treatment in neuroblastoma cells, the proteinexpression levels of ER stress markers ATF5 andphosphorylated eIF2 did not display any change whencompared to control cells. We also showed that ATP levelswere decreased in kainic acid-treated cells.Conclusion: This study may show that the level of stressthat kainic acid causes at 1 mM for 24 hours inneuroblastoma cells was not adequate to lead to ER stresswhich is measurable by ATF5 and phosphorylated eIF2.Either an increased level of treatment of kainic acid viaincreased duration or concentration is necessary ordifferent markers should be tried. The investigation of theER stress pathways in the excitotoxicity-related braindiseases will pave the way for new therapies based on ERstress and combat more than one disease simultaneously

Anahtar Kelime:

Endoplazmik retikulum stres belirteçlerinden ATF5 ve fosforile IF2 düzeylerinin nöroblastoma hücrelerinde kainik asit muamelesi sonrası incelenmesi

Öz:

Amaç: Bu çalışmada, amacımız, kainik asite bağlı eksitotoksisite ve endoplazmik retikulum (ER) stres arasındaki ilişkiyi iki majör endoplazmik retikulum stres markırı olan ATF5 ve fosforile olmuş eIF2a analiz ederek incelemektir. Gereç ve Yöntem: Neuroblastoma hücrelerini 1 mM kainik asit ile 24 saat muamele ettik. ATP ölçümü, kainik asit muamelesi yapılan hücrelerde ya da kontrol hücrelerinde bioluminesans bir yöntem ile yapılmıştır. Total protein, kainik asit muamelesi yapılmış ya da kontrol hücrelerinden izole edilmiş ve ATF5 ve fosforile olmuş eIF2 markırları western blot ile incelenmiştir. Bulgular: Kainik asit ile muamele edilmiş neuroblastoma hücrelerinde ATF5 ve fosforile olmuş eIF2a seviyelerinin kontrole göre değişmediğini ilk kez gösterdik. Kainik asit ile muamele edilmiş hücrelerde ATP seviyesinin düştüğünü gösterdik. Sonuç: 1 mM ve 24 saat süren kainik asit muamelesi, ATF5 ve fosforile olmuş eIF2 ile gösterilebilecek endoplazmik retikulum stresi yaratmak için yeterli olmayabilir. Süre ve konsantrasyon olarak arttırılmış kainik asit muamelesi ya da farklı markırlar gerekmektedir. Eksitotoksisiteye bağlı beyin hastalıklarında ER stres yolaklarını araştırmak, yeni tedavi yolları bulmak ve birden fazla hastalığı aynı anda engellemek adına önemli olacaktır.

Anahtar Kelime:

Belge Türü: Makale Makale Türü: Araştırma Makalesi Erişim Türü: Erişime Açık

1. Oakes SA, Papa FR. the role of endoplasmic reticulum stress in human pathology. Annu Rev Pathology. 2015;10:173-94.
2. Hetz C. The unfolded protein response: controlling cell fate decisions under ER stress and beyond. Nature Rev Mol Cell Biol. 2012;13:89-102.
3. Adams CJ, Kopp MC, Larburu N, Nowak PR, Ali MMU. Structure and molecular mechanism of ER stress signaling by the unfolded protein response signal activator IRE1. Front Mol Biosci. 2019;6:11.
4. Liu CY, Kaufman RJ. The unfolded protein response. J Cell Sci. 2003;116:1861-2.
5. Pakos-Zebrucka K, Koryga I, Mnich K, Ljujic M, Samali A, Gorman AM. The integrated stress response. EMBO Rep. 2016;17:1374–95.
6. Torres-Peraza JF, Engel T, Martin-Ibanez R, SanzRodriguez A, Fernandez-Fernandez MR, Esgleas M, et al. Protective neuronal induction of ATF5 in endoplasmic reticulum stress induced by status epilepticus. Brain. 2013:136;1161-76.
7. Dong X, Wang Y, Qin Z. Molecular mechanisms of excitotoxicity and their relevance to pathogenesis of neurodegenerative diseases. Acta Pharmacol Sin. 2009;30:379-87.
8. Zhou Y, Danbolt NC. Glutamate as a neurotransmitter in the healthy brain. J Neural Transm. 2014;121:799-817.
9. Olivares-Bañuelos TN, Chí-Castañeda D, Ortega A. Glutamate transporters: Gene expression regulation and signaling properties. Neuropharmacology. 2019;161:107550.
10. Karaca M, Frigerio F, Maechler P. From pancreatic islets to central nervous system, the importance of glutamate dehydrogenase for the control of energy homeostasis. Neurochem Int. 2011;59:510-7.
11. Danbolt NC. Glutamate uptake. Prog Neurobiol. 2001;65:1-105.
12. Lin CLG, Kong Q, Cuny GD, and Glicksman MA. Glutamate transporter EAAT2: a new target for the treatment of neurodegenerative diseases. Future Med Chem. 2012;4:1689-700.
13. Mohd Sairazi NS, Sirajudeen KNS, Asari MA, Muzaimi M Mummedy S, Sulaiman SA. Kainic acidinduced excitotoxicity experimental model: protective merits of natural products and plant extracts. Evid Based Complement Alternat Med. 2015;2015:972623.
14. Verdaguer E, Garcia-Jorda E, Jimenez A, Stranges A, Sureda FX, Canudas AM et al. Kainic acid-induced neuronal cell death in cerebellar granule cells is not prevented by caspase inhibitors. Br J Pharmacol. 2002;135:1297-307.
15. Vermoesen K, Smolders I, Massie A, Michotte Y, Clinckers R. The control of kainic acid-induced status epilepticus. Epilepsy Res. 2010;1-2:164-6.
16. Cherian A, Thomas SV. Status epilepticus. Ann Indian Acad Neurol. 2009;12:140–53.
17. Sokka AL, Putkonen N, Mudo G, Pryazhnikov E, Reijonen S, Khiroug L et al Endoplasmic reticulum stress inhibition protects against excitotoxic neuronal injury in the rat brain. J Neurosci. 2007;27:901-8.
18. Xue F, Shi C, Chen Q, Hang W, Xia L, Wu Y et al. Melatonin mediates protective effects against kainic acid-induced neuronal death through safeguarding er stress and mitochondrial disturbance. Front Mol Neurosci. 2017;10:49.
19. Kim JS, Heo RW, Kim H, Yi CO, Shin HJ, Han JW et al. Salubrinal, ER stress inhibitor, attenuates kainic acid-induced hippocampal cell death. J Neural Transm. 2014;121:1233.
20. Zhang XM, Zhu J. Kainic acid-induced neurotoxicity: targeting glial responses and glia-derived cytokines. Curr Neuropharmacol. 2011;9:388–98.
21. Zheng X, Zhang HL, Luo Q, Zhu J. Kainic acidinduced neurodegenerative model: potentials and limitations. J Biomed and Biotech. 2010;2011:457079. 22. Yoshida H. ER stress and diseases. FEBS J. 2007;274:630-58.

APA	SARI A, Dönmez Yalçın G (2020). The investigation of endoplasmic reticulum stress markers ATF5 and phosphorylated eIF2 after kainic acid treatment in neuroblastoma cells. , 96 - 101. 10.17826/cumj.637075
Chicago	SARI Alime,Dönmez Yalçın Gizem The investigation of endoplasmic reticulum stress markers ATF5 and phosphorylated eIF2 after kainic acid treatment in neuroblastoma cells. (2020): 96 - 101. 10.17826/cumj.637075
MLA	SARI Alime,Dönmez Yalçın Gizem The investigation of endoplasmic reticulum stress markers ATF5 and phosphorylated eIF2 after kainic acid treatment in neuroblastoma cells. , 2020, ss.96 - 101. 10.17826/cumj.637075
AMA	SARI A,Dönmez Yalçın G The investigation of endoplasmic reticulum stress markers ATF5 and phosphorylated eIF2 after kainic acid treatment in neuroblastoma cells. . 2020; 96 - 101. 10.17826/cumj.637075
Vancouver	SARI A,Dönmez Yalçın G The investigation of endoplasmic reticulum stress markers ATF5 and phosphorylated eIF2 after kainic acid treatment in neuroblastoma cells. . 2020; 96 - 101. 10.17826/cumj.637075
IEEE	SARI A,Dönmez Yalçın G "The investigation of endoplasmic reticulum stress markers ATF5 and phosphorylated eIF2 after kainic acid treatment in neuroblastoma cells." , ss.96 - 101, 2020. 10.17826/cumj.637075
ISNAD	SARI, Alime - Dönmez Yalçın, Gizem. "The investigation of endoplasmic reticulum stress markers ATF5 and phosphorylated eIF2 after kainic acid treatment in neuroblastoma cells". (2020), 96-101. https://doi.org/10.17826/cumj.637075

APA	SARI A, Dönmez Yalçın G (2020). The investigation of endoplasmic reticulum stress markers ATF5 and phosphorylated eIF2 after kainic acid treatment in neuroblastoma cells. Cukurova Medical Journal, 45(1), 96 - 101. 10.17826/cumj.637075
Chicago	SARI Alime,Dönmez Yalçın Gizem The investigation of endoplasmic reticulum stress markers ATF5 and phosphorylated eIF2 after kainic acid treatment in neuroblastoma cells. Cukurova Medical Journal 45, no.1 (2020): 96 - 101. 10.17826/cumj.637075
MLA	SARI Alime,Dönmez Yalçın Gizem The investigation of endoplasmic reticulum stress markers ATF5 and phosphorylated eIF2 after kainic acid treatment in neuroblastoma cells. Cukurova Medical Journal, vol.45, no.1, 2020, ss.96 - 101. 10.17826/cumj.637075
AMA	SARI A,Dönmez Yalçın G The investigation of endoplasmic reticulum stress markers ATF5 and phosphorylated eIF2 after kainic acid treatment in neuroblastoma cells. Cukurova Medical Journal. 2020; 45(1): 96 - 101. 10.17826/cumj.637075
Vancouver	SARI A,Dönmez Yalçın G The investigation of endoplasmic reticulum stress markers ATF5 and phosphorylated eIF2 after kainic acid treatment in neuroblastoma cells. Cukurova Medical Journal. 2020; 45(1): 96 - 101. 10.17826/cumj.637075
IEEE	SARI A,Dönmez Yalçın G "The investigation of endoplasmic reticulum stress markers ATF5 and phosphorylated eIF2 after kainic acid treatment in neuroblastoma cells." Cukurova Medical Journal, 45, ss.96 - 101, 2020. 10.17826/cumj.637075
ISNAD	SARI, Alime - Dönmez Yalçın, Gizem. "The investigation of endoplasmic reticulum stress markers ATF5 and phosphorylated eIF2 after kainic acid treatment in neuroblastoma cells". Cukurova Medical Journal 45/1 (2020), 96-101. https://doi.org/10.17826/cumj.637075