ALS Sinek Modelinde Nörodejenerasyon Mekanizmasının Proteomik ve Genetik Analizi
Amaç : TDP-43; RNA bağlanma motifleri içeren ve pre-mRNA ekleme, transkripsiyon, mRNA stabilitesive mRNA transferinde yer aldığı rapor edilen oldukça korunmuş ve her yerde eksprese edilennükleer proteindir. Son zamanlarda TDP-43 yapılan çalışmalar ile Amyotrofik Lateral Skleroz (ALS)hastalarının etkilenmiş beyin bölgelerinde gözlenen hücre içi inklüzyonların ana protein bileşeni olaraktanımlandı. Bu çalışmada Drosophila melanogaster’de TDP-43’ün analoğunun (TBPH’in) ALS modelindekisinekler üzerinde proteomik ve genetik mekanizmaları içerisindeki rolünün araştırılması amaçlanmıştır.Materyal-Metod : WIII8,UAS GFP, GMR,GMR TBPH genotiplerinde bakire dişi sinekler ve erkekleruygun besiyeri ortamında çiftleştirilerek farklı genotip ve fenotiplerde ALS için etki gösterebilentransgenik model sinekler üretildi. Transgenik sineklere TBPH entegre edildi. PCR , Jel Elektroforezi ve2D kütle spektrofotometri yöntemleri kullanılarak TBPH mutantlarının analizi yapıldı.Bulgular : TDP-43’ün analoğu olan TBPH’i eksprese eden farklı transgenik sinekler kullanılarak ,TBPH’in ekspresyonunun kontrol gruplarına kıyasla nörodejenerasyona neden olup olmadığı kontroledildi. TPBH’nin ekspresyonunun sineklerin gözlerinde siyah lekelerin (nekrozun) gelişmesine nedenolduğu ve gözlerde nörodejenerasyon oluşturduğu görüldü. Dahası, mutasyon geçirmiş sinekler üzerinde2D jel analizi ile protein agregasyonunda TBPH’in rolünü düşündüren sinek türlerine kıyasla diğersineklerin gözlerinde azalmış yağ vücut proteinleri 1 (Fbp1) seviyeleri gösterildi. Sonuçlar : Sonuç olarak TBPH’in Fbp1’nın üretimini artırarak nörodejeneratif sürece karıştığı görüldü.
Anahtar Kelimeler:
ALS, Drosophila, TDP-43, Fbp1
Proteomic and Genetic Analysıs of the Mechanism of Neurodegeneration in a Fly Model of ALS
Objective : TDP-43 is a highly conserved and ubiquitously expressed nuclear protein containingRNA binding motives and reported to be involved in pre-mRNA splicing, transcription, mRNA stabilityand mRNA transport. Recently, TDP-43 was identified as the main protein component of the intracellularinclusions observed in affected brain areas of patients suffering from Amyotrophic Lateral Sclerosis(ALS).Material and Methods : In WIII8, UAS GFP, GMR, GMR TBPH genotypes, virgin and male transgenicmodel flies were produced which were able to act for ALS in different genotypes and phenotypes bymating them in the appropriate medium. TBPH was integrated into the transgenic flies. TBPH mutantswere analyzed using PCR, Gel Electrophoresis and 2D mass spectrophotometry methods.Results : We have used different transgenic flies expressing TDP-43 analogue TBPH and checkedwhether the expression of TBPH is causing the neurodegeneration as compared to the control groups.Expression of TPBH caused development of black spot (necrosis) in the eyes of the flies clearly suggestedneurodegeneration in the eye. Moreover, 2D gel analysis on TBPH mutated flies showed reducedfat body proteins 1 (Fbp1) levels in the eyes of flies as compared to wild type, suggesting the role ofTBPH in the protein aggregation.Conclusion : Our date showed TBPH is involved in the neurodegenerative process by enhancing the production of Fbp11.
Keywords:
ALS Drosophila, TDP-43, TBPH, Fbp1,
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- 1. Movement Dısorders Genetics and Models ; Mark S.LeDoux -Gama Sosa et. all 535-536 London : Elsevier/Academic Press, [2015] , YOUSUF ALI KAIRUAN,R.GRACE ZHAI; In IX- .TAUOPATHIES, Drosophila Model of Tautopathy 830-845 2. From charcot to lou gehrig: deciphering selective motor neuron death in ALS Don W. Cleveland1 & Jeffrey D. Rothstein2 Nature Reviews Neuroscience 2, 806-819 (November 2001) | doi:10.1038/35097565 3. Motor Neuron Disease Research Progress ; Raffaele L.Mancini -Worms et. all 188-189 ISBN : 978-1-60456-155-5 ,1.Amyotrophic Lateral Sclerosis. 2.Motor Neurons – Diseases 131-159 4. Protein Misfolding in Neurodegenerative Diseases ; Mechanisms and Therapeutic Strategies H.John Smith , Claire Simons , Robert D.E.Sewell London-New York , Veltema et all 37-39 5. Motor Neuron Disease : Biology and Management P. N. Leigh and M.Swash In British Library Catologuing in Publication Data UK ISBN : 13:978-1-4471-1871-8, Piao et all 487 6. Superoxide Dismutase Activity, Oxidative Damage, and Mitochondrial Energy- Metabolism in Familial and Sporadic Amyotrophic Lateral Sclerosis Authors-Allen C. Bowling,- Jorg B. Schulz,Robert H. Brown Jr.,M. Flint Beal-First published: December 1999 7. Odification of Superoxide Dismutase 1 (SOD1) Properties by a GFP Tag – Implications for Research into Amyotrophic Lateral Sclerosis (ALS)- James C. Stevens ,Ruth Chia ,William T. Hendriks,Virginie Bros-Facer,Jan van,Minnen, Joanne E. Martin,Graham S. Jackson,Linda Greensmith,Giampietro Schiavo,Elizabeth M. C. Fisher - Published: March 8, 2010 8. The Microglial-Motoneuron dialogue in ALS S.H. Appel, W. Zhao, D.R. Beers, J.S. Henkel Methodist Neurological Institute, Methodist Hospital Research Institute, Houston, TX USA- Acta Myologica • 2011; XXX: p. 4-8 9. Cortical hyperexcitability may precede the onset of familial amyotrophic lateral sclerosis- Steve Vucic, Garth A. Nicholson, Matthew C. Kiernan-Brain (2008) 131 (6): 1540- 1550. Published: 09 May 2008 10. Neurol Neurosurg Psychiatry 2007;78:849- 852 doi:10.1136/jnnp.2006.105056-Abnormalities in cortical and peripheral excitability in flail arm variant amyotrophic lateral sclerosis , Steve Vucic1, Matthew C Kiernan2 11. Ultrastruct Pathol. 2002 Jan-Feb;26(1):3-7. Ultrastructural changes of mitochondria in the skeletal muscle of patients with amyotrophic lateral sclerosis.- Chung MJ1, Suh YL 12. Review-Mitochondrial dysfunction in amyotrophic lateral sclerosis, Ping Shia, Jozsef Galb, David M. Kwinterb, Xiaoyan Liub, Haining Zhua, ,January 2010, Pages 45–51, Mitochondrial Dysfunction 13. Muscle Nerve. 2016 Jun;53(6):842-9. doi: 10.1002/mus.25049. Epub 2016 Apr 8.- Accrued somatic mutations (nucleic acid changes) trigger ALS: 2005-2015 update. Armon C1. 14. J Neurol Sci. 2010 Jan 15;288(1-2):1-12. doi: 10.1016/j.jns.2009.09.029. Epub 2009 Oct18. 15. The evidence for altered RNA metabolism in amyotrophic lateral sclerosis (ALS) -Strong MJ1 16. Antioxid Redox Signal. 2009 Jul; 11(7): 1587–1602. Glutamate Transporters and the Excitotoxic Path to Motor Neuron Degeneration in Amyotrophic Lateral Sclerosis- Emily Foran and Davide Trotti 17. A Controlled Trial of Riluzole in Amyotrophic Lateral Sclerosis -G. Bensimon, L. Lacomblez, V. Meininger, and *the ALS/Riluzole Study Group -N Engl J Med 1994; 330:585-591March 3, 1994 18. Cloning and characterization of a novel cellular protein, TDP-43, that binds to human immunodeficiency virus type 1 TAR DNA sequence motifs. S H Ou, F Wu,D Harrich,L F García-Martínez,R B Gaynor- J. Virol. June 1995 vol. 69 no. 63584-3596 19. The International Journal of Biochemistry & Cell Biology - Volume 42, Issue 10, October 2010, Pages 1606–1609 , TDP-43: A DNA and RNA binding protein with roles in neurodegenerative diseases- Sadaf T. Warraicha, b, Shu Yanga, Garth A. Nicholsona, b, c, Ian P. Blaira, b 20. Journal of Neurology , August 2009, Volume 256, Issue 8, pp 1205–1214 , Amyotrophic lateral sclerosis, frontotemporal dementia and beyond: the TDP-43 diseases , Felix Geser , Maria Martinez-Lage,Linda K. Kwong,- Virginia M.-Y. Lee,John Q. Trojanowski 21. Depletion of TDP-43 affects Drosophila motoneuronsterminal synapsis and locomotive behavior Volume 583, Issue 10 , May 19, 2009 , Pages 1586–1592 , Fabian Feiguin,Vinay K. Godena,Giulia Romano,Andrea D’Ambrogio,Raffaella Klima,Francisco E. Baralle 22. TDP-43 Regulates Drosophila Neuromuscular Junctions Growth by Modulating Futsch/ MAP1B Levels and Synaptic Microtubules Organization , Vinay K. Godena ,Giulia Romano ,Maurizio Romano,Chiara Appocher,Raffaella Klima,Emanuele Buratti,Francisco E. Baralle ,Fabian Feiguin 23. DROSOPHİLA- Therese A.Markow,Patrick M. O’Grady , Academic Press (December 7, 2005) , ISBN-10: 0124730523, ISBN-13: 978- 0124730526
- Yayın Aralığı: Yılda 3 Sayı
- Başlangıç: 2011
- Yayıncı: İstanbul Üniversitesi
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