Determining of Apoptosis at in vitro Conditioned DRG Neurons with TUNEL Method

Yıl 2017, Cilt: 7 Sayı: 2, 80 - 84, 26.12.2017

Elif Kaval Oğuz

https://doi.org/10.17678/beuscitech.331471

Öz

In this study, it was aimed to prove the way of in
vitro conditioned cells death. There is an increased axonal regeneration after a
peripheral nerve is injured if it exposed to an earlier prior injury. The first
injury causing the enhanced regeneration is called a conditioning lesion. In
this study, Dorsal Root Ganglion primer cell culture was performed then neurons
were conditioned in vitro with an effective and new method. It was observed that in
vitro conditioned cells were regenerated much faster than control group cells
but at the end of 48^th hour, they died. The cell death way of these
in vitro conditioned neurons was determined by an in situ Cell Detection Kit Fluorescein and visualized by
fluorescent microscopy. Finally, apoptotic cell deaths observed at in vitro conditioned neurons. This study
is important for the continuity of enhanced regeneration capacity of
conditioning lesion effect and neuronal survival and also it will provide the
basis for new studies.

Anahtar Kelimeler

Dorsal root ganglion (DRG), regeneration, conditioning lesion

Kaynakça

• Bastian, A. M., Yogesh T. L., Kuraswamy, K. L., 2013. Various Methods Available for Detection of Apoptotic Cells- A Review. Indian Journal of Cancer. 50 (3): 274-283.
• Bouchier-Hayes, L., Munoz-Pinedo, C., Connell, S., Green, D.,R., 2008. Measuring Apoptosis at the single level. Methods. (44): 222-228.
• Bredesen, D. E., Rao, R. V., Mehlen, P., 2006. Cell Death in the Nervous System. Nature. 443(7113): 796-802.
• Cafferty, W. B. J., Gardiner, N. J., Das, P., Qui, J., McMahon, S. B., Thompson, S. W. N., 2004. Conditioning ınjury-induced spinal axon regeneration fails in Interleukin-6 knock-out mice. The Journal of Neuroscience. 24, (18): 4432-4443.
• Cafferty, W. B. J., Gardiner, N. J., Gavazzi, I., Powell, J., McMahon, S. B., Heath, J. K., Munson, J., Cohen, J., Thompson, S. W. N., 2001. Leukemia Inhibitory Factor determines growth status of injured adult sensory neurons. The Journal of Neuroscience. 21, (18): 7161-7170.
• Cengiz, N., Öztürk, G., Erdoğan, E., Him, A., Kaval Oğuz, E., 2012. Consequences of Neurite Transection In Vitro. Journal of neurotrauma: (29):2465-2474. Cooper, G. M., Hausman, R. E., 2006. Hücre Moleküler Yaklaşım, 3. Baskı. İzmir Tıp Kitabevi. İzmir. 579-582.
• Dahlin, L., B., 2004. The biology of nerve injury and repair. Journal of The American Society For Surgery of The Hand. (3):143-155.
• Doron-Mandel, E., Fainzilber, M., Terenzio, M., 2015. Growth Control Mechanisms in Neuronal Regeneration. FEBS Letters. 589(14), 1669-1677.
• Edström, A., Ekström P. A., Tonge, D. 1996. Axonal Outgrowth and Neuronal Apoptosis in Cultured Adult Mouse Dorsal Root Ganglion Preparations: Effects of Neurotrophins of Inhibition of Neurotrophin Actions and of Prior Axotomy. Neurocience. 75, 1165-1174.
• Elmore, S., 2007. Apoptosis: A Review of Programmed Cell Death. Toxicol Pathol. 35(4): 495-516.
• Geçit, İ., Kavak, S., Kaval Oğuz, E., Pirincci, N., Güneş, M., Kara, M., Ceylan, K., Kaba, M., Tanık, S., 2014. Tissue Damage in Kidney, Adrenal Glands and Diaphram Following Extracorporeal Shock Wave Lithotripsy. Toxicoly and Industrial Health. 30 (9): 845-850.
• Guimaraes C. A., Linden R., 2004. Programmed Cell Death Apoptosis and Altenative Deathstyles. Eur. J. Biochem. 271: 1638-1650.
• Hu-Tsai, M., Winter, J., Emson, P. C., Woolf, C., 1994. Neurite outgrowth and GAP 43 expression in cultured adult rat dorsal root ganglion neurons: effects of NGF or prior peripheral axotomy. J. Neurosci. Res. (39): 634-645.
• Kaval Oğuz, E., 2009. Searching of Conditioning Lesion Effect and Its Mechanism In Nerve Regeneration. PhD Thesis. Yüzüncü Yıl University Van. 187.
• Koff, J. L., Ramachandiran, S., Bernal-Mizrachi, L., 2015. A time to Kill: Targeting Apoptosis in Cancer. Int. J. Mol. 16: 2942-2955.
• Krysko, D. V., Berghe, T.V., D’herde, K., Vandenabeele, P., 2008. Apoptosis and Necrosis: Detection, discrimination and phagocytosis. Methods. (44): 205-221.
• Liu., R. Y., Snider, W.D., 2001. Different signalling pathways mediate regenerative versus developmental sensory axon growth. J. Neurosci. (21): RC164
• Lüleyap, H. Ü., 2008. Moleküler Genetiğin Esasları, Nobel Kitabevi. Adana. 292-299.
• McQuarrie, I. G., Grafstein, B., 1973. Axon Outgrowth Enhanced by a previous Nerve Injury. Arch. Neurol. 29, 53-55.
• Qui, J., Cafferty, W. B., McMahon, S. B., Thompson, S. W., 2005. Conditioning injury-induced spinal axon regeneration requires signal transducer and activator of transcription 3 activation. J. Neurosci. (25): 1645-1653.
• Sachs, H. H., Rohrer, H., Zigmond, R. E., 2010. The Conditioning Lesion Effect on Sympathetic Neurite Outgrowth is Dependent on gp130 Cytokines. Experimental Neurology. 223: 516-522.
• Shoemaker, S. E., Sachs, H. H., Vaccariello, S. A., Zigmond, R. E. 2005. A Conditioning Lesion Enhances Sympathetic Neurite Outgrowth. Exp. Neurol. 194, 432-443.
• Shoemaker, S. E., Sachs, H. H., Vaccariello, S. A., Zigmond, R. E., 2006. Reduction in nerve growth factor availability leads to a conditioning lesion-like effect in sympathetic neuronsJournal of Neurobiolgy. (66): 1322-1337.
• Thornberry, N. A., Lazebnik, Y., 1998. Caspases: enemies within. Science. 281: 1312-1316.
• Ulukaya, E., Acilan, C., Ari, F., İkitimur, E., Yilmaz, Y. A., 2011. Galance at the Methods for Detection of Apoptosis Qualitatively and Quantatively. Turk. J. Biochem. 36(3); 261-269.
• Van Cruchten, S., Van Den Broeck, W., Morphological and Biochemical Aspects of Apoptosis, Oncosis an Necrosis. 2002. Anat. Histol. Embryol. 31; 214-223.
• Zhang, L., Palmer, R., McClellan, A. D., 2004. Conditioning lesion enhance axonal regeneration of descending brain neurons in spinal-cord-transected larval lamprey. The Journal of Comparative Neurology. (478): 395-404.
• Ziegler, U., Groscurth, P., 2004. Morphological Features of Cell Death. News Physol. Sci. 19: 124-128.