Apoptozun Belirlenmesinde Kullanılan Yöntemler

Öz

Anahtar Kelimeler

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Apoptozun Belirlenmesinde Kullanılan Yöntemler

Öz

Apoptoz (fizyolojik hücre ölümü), hücre intiharı olarak da bilinen fizyolojik bir olaydır. Apoptoz için sinyal alındıktan sonra hücrede birçok biyokimyasal (DNA parçalanması, fosfotidilserin moleküllerinin hücre membranın dış yüzeyine çıkması) ve morfolojik değişim (hücre küçülmesi, kromatin yoğunlaşması, apoptotik cisimcikler) gözlenir. Bu değişimleri saptamak için çok çeşitli yöntemler geliştirilmiştir. DNA fragmentasyonu histokimyasal olarak (TUNEL) gösterilebileceği gibi, biyokimyasal (agoroz jel elektroforezi) olarak da gösterilebilir. Hatta, ELISA ile de gösterilebilir. Transmisyon elektron mikroskopisi, apoptozun morfolojik olarak tanınması için iyi bir yöntemdir. Apoptoz ayrıca Caspase-3 (Kaspaz-3) metodu kullanılarak da belirlenebilir

Anahtar Kelimeler

• Apoptoz, Fizyolojik Hücre Ölümü, DNA Parçalanması, TUNEL

Kaynakça

1. Aral H (1996): Apoptotik hücrelerin tanınma yöntemleri. Sendrom, 38-42.
2. Bardakçı F, Yenidünya A F (2007): Moleküler biyoloji teknikleri 1:Nükleik asit analiz teknikleri (in) Moleküler Biyoloji. A Yıldırım, F Bardakçı, M Karataş, B Tanyolaç (Editör), 519-553, Nobel Yayın, Ankara.
3. Bellamy C O, Malcomson, R D, Harrison, D J, Wyllie A H (1995): Cell death in health and disease: the biology and regulation of apoptosis. Cancer Biology, 6: 3-16.
4. Bortner C D, Oldenburg N B, Cidlowski J A (1995): The role of
5. DNA fragmentation in apoptosis. Trends Cell Biol., 5: 21–6.
6. Bossù P (2007): Qualitative analysis of dna fragmentation by agarose [http://www.cyto.purdue.edu/flowcyt/research/cytotech/ap opto/data/chap4.htm] Erişim Tarihi: 05.11.2007. Erişim:
7. Bratton D L, Fadok V A, Richter D A, Kailey J M, Guthrie L A,
8. Henson P M (1997): Appearance of phosphatidylserine on

9. apoptotic cells requires calcium-mediated nonspecific flip
10. flop and is enhanced by loss of the aminophospholipid
11. translocase. J. Biol. Chem., 272: 26159–26165.
12. Carson D A, Rbiero J M (1993): Apoptosis and disease. The Lancet, 341: 1251-1254.
13. Caulin C, Salvesen G S, Oshima R G (1997): Caspase cleavage
14. of keratin 18 and reorganization of ıntermediate filaments
15. during epithelial cell apoptosis. The Journal of Cell Biology, 138(6): 1379-1394.
16. Cohen J J (1993): Programmed cell death and apoptosis in lymphocyte development and function. American College of Physicians, CHEST, 103: 99-101. Cummings M C, Winterfold C M, Walker N I (1997): Apoptosis. Am. J. Surg. Pathol., 21: 88-101.
17. Elmore S (2007) Apoptosis: A review of programmed cell death. Toxicol. Pathol., 35(4): 495–516.
18. Gatti R, Belletti S, Orlandini G, Bussolati O, Dall’asta V, Gazzola G C (1998): Comparison of Annexin V and calcein-AM as early vital markers of apoptosis in adherent cells by confocal laser microscopy. J. Histochem. Cytochem., 46: 895–900.
19. Gavrieli Y, Sherman Y, Ben-Sasson S A (1992): Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. J. Cell Biol., 119: 493-501.
20. Grieken N C T, Meijer G A, Hausen A, Meuwissen S G M, Baak J P A, Kuipers E J (2003): Increased apoptosis in gastric mucosa adjacent to intestinal metaplasia. J. Clin. Pathol., 56: 358–362.
21. Kockx M M, Muhring J, Knaapen M W M, de Meyer G R Y (1998): RNA synthesis and splicing interferes with DNA in situ end labeling techniques used to detect apoptosis. Am. J. Pathol., 152: 885.
22. Kopman G, Reutelingsperger C P, Kuijten G A, Keehnen R M, Pals S T, van Oers N H (1994): Annexin V for flow cytometric detection of phosphatidylserine expression on B cells undergoing apoptosis. Blood., 84: 1415–1420.
23. Kressel M, Groscurth P (1994): Distinction of apoptotic and necrotic cell death by in situ labelling of fragmented DNA. Cell Tissue Res., 278: 549–56.
24. Leers M P, Kolgen W, Bjorklund V, Bergman T, Tribbick G, Persson B, Bjorklund P, Ramaekers F C, Bjorklund B, Nap M, Jornvall H, Schutte B (1999): Immunocytochemical detection and mapping of a cytokeratin 18 neo-epitope exposed during early apoptosis. J. Pathol., 187: 567–572.
25. Majno G, Joris I (1995): Apoptosis, oncosis, and necrosis: An overview of cell death. Am. J. Pathol., 146: 3-15.
26. Mak T (2003): The E. Donnall Thomas Lecture-apoptosis: "tis death that makes life live". Biol. Blood Marrow Transplant, 9: 483-8.
27. Morrone S (2007): Annexın V. Purdue Cytometry CD-ROM Series, volume 4 Erişim: [http://www.cyto.purdue.edu/ flowcyt/research/cytotech/apopto/data/chap16.htm] Erişim Tarihi: 02.12.2007.
28. Mountz J D, Zhou T (2001): Apoptosis and autoimmunity (in) A Textbook of Rheumatology: Artritis and allied conditions. WJ Kopman (Editör), Lippincott-Williams&Wilkins.
29. Overbeeke R, Steffens-Nakken H, Vermes I, Reutelingsperger C, Hanen C. (1998): Early features of apoptosis detected by four different flow cytometry assays. Apoptosis, 3: 115.
30. Öniz H (2004): Apoptoz: Ölmeye yatmak. SSK Tepecik Eğitim Hastanesi Dergisi, 14(1).
31. Palazzo I G, Lega A, Farinha G S, Berrotarán N, Ortiz S (2005): de presentacion de un caso. Congreso Virtual Hispanoamericano de Anatomia Patologica. utero:linfoma primario,
32. Pınarbaşı E (2007): Apoptozis (Programlı Hücre Ölümü) (in) Moleküler Biyoloji. A Yıldırım, F Bardakçı, M Karataş, B Tanyolaç (Editör), 423-468, Nobel Yayın, Ankara.
33. Sakahira H, Enari M, Nagata S (1998): Cleavage of CAD inhibitor in CAD activation and DNA degradation during apoptosis. Nature., 391: 96–9.
34. Salgame P, Varadhachary A S, Primiano L L, Fincke J E, Muller S, Monestier M (1997): An ELISA for detection of apoptosis. Nucleic Acids Research, 25(3): 680–681.
35. Schwartzman R A, Cidloski J A (1993): Apoptosis; the biochemistry and molecular biology of programmed cell death. Endocrine Reviews, 14: 133-144.
36. Sereno D, Holzmuller P, Mangot I, Cuny G, Ouaissi A, Lemesre J L (2001): Antimonial-mediated dna fragmentation in leishmania infantum Chemother, 45(7): 2064–2069.
37. Spector D L, Goldman R D, Leinwand L A (1998): Culture and Biochemical Analysis of Cells (in) Cells: A Laboratory Manual. DL Spector (Editör), 15.1-15.24, Cold Spring Harbor Laboratory Press, USA.
38. Squier M K, Miller A C, Malkinson A M, Cohen J J (1994): Calpain activation in apoptosis. J. Cell Physiol., 159(2): 229- 237.
39. Tesarik J, Greco E, Cohen-Bacrie P, Mendoza C (1998): Germ cell apoptosis in men with complete and incomplete spermiogenesis failure. Mol. Hum. Reprod., 4: 757.
40. Thompson C B (1995): Apoptosis in the pathogenesis and treatment of disease. Science, 267: 1456–1462. Trump B F, Berezesky I K, Chang S H, Phelps P C (1997): The pathways of cell death: oncosis, apoptosis, and necrosis. Toxicol. Pathol., 25: 82–8.
41. Ulukaya E (2003): Apoptozis Ders Notları. Erişim: [http://biyokimya.uludag.edu.tr/ Erişim tarihi: 04.05.2007. Vaux D L, Flavell R A (2000): Apoptosis genes and autoimmunity. Curr. Opin. Immunol., 12: 719–724.