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. 2008 Jul 19;24(3):143–149. doi: 10.1007/s12264-008-1108-0

Autophagy is activated and might protect neurons from degeneration after traumatic brain injury

大鼠脑外伤后自噬被激活并在早期对受损神经元起保护作用

Yan-Bo Zhang 1, Sheng-Xing Li 1, Xi-Ping Chen 1, Li Yang 1, Yun-Ge Zhang 1, Ran Liu 1, Lu-Yang Tao 1,✉
PMCID: PMC5552539  PMID: 18500386

Abstract

Objective

To investigate changes of autophagy after traumatic brain injury (TBI) and its possible role.

Methods

Rat TBI model was established by controlled cortical injury system. Autophagic double membrane structure was detected by transmission electronic microscope. Microtubule-associated protein 1 light chain 3 (LC3) and Beclin 1 were also used to investigate the activation of autophagy post-TBI. Double labeling with LC3 and caspase-3, or Beclin 1 and Fluoro-Jade, to show the relationship between autophagy and apoptosis or neuron degeneration after TBI.

Results

An increase of autophagic double membrane structure was observed in early stage (1 h), and the increase lasted for at least 32 d post-TBI. LC3 and Beclin 1 proteins also began to elevate at 1 h time point post-TBI in neurons, 3 d later in astrocytes, and peaked at about 8 d post-TBI. In both cell types, LC3 and Beclin 1 maintained at a high level until 32 d post-TBI. Most LC3 and Beclin 1 positive cells were near the side (including hippocampus), but not in the core of the injury. In addition, in the periphery of the injury site, not all caspase-3 positive (+) cells merged with LC3 (+) cells post-TBI; In hippocampal area, almost all Beclin 1 (+) neurons did not merge with Fluoro-Jade (+) neurons from 1 h to 48 h post-TBI.

Conclusion

Autophagy is activated and might protect neurons from degeneration at early stage post-TBI and play a continuous role afterwards in eliminating aberrant cell components.

Keywords: autophagy, apoptosis, traumatic brain injury, LC3, Beclin1, neurodegeneration

Footnotes

The two authors contributed equally to this work.

References

  • [1].Mizushima N., Ohsumi Y., Yoshimori T. Autophagosome formation in mammalian cells. Cell Struct Funct. 2002;27:421–429. doi: 10.1247/csf.27.421. [DOI] [PubMed] [Google Scholar]
  • [2].Cuervo A.M. Autophagy: in sickness and in health. Trends Cell Biol. 2004;14:70–77. doi: 10.1016/j.tcb.2003.12.002. [DOI] [PubMed] [Google Scholar]
  • [3].Levine B., Klionsky D.J. Development by self-digestion: molecular mechanisms and biological functions of autophagy. Dev Cell. 2004;6:463–477. doi: 10.1016/S1534-5807(04)00099-1. [DOI] [PubMed] [Google Scholar]
  • [4].Kegel K.B., Kim M., Sapp E., McIntyre C., Castano J.G., Aronin N., et al. Huntingtin expression stimulates endosomal-lysosomal activity, endosome tubulation, and autophagy. J Neurosci. 2000;20:7268–7278. doi: 10.1523/JNEUROSCI.20-19-07268.2000. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [5].Zakeri Z., Bursch W., Tenniswood M., Lockshin R.A. Cell death: programmed, apoptosis, necrosis, or other? Cell Death Diff. 1995;2:87–96. [PubMed] [Google Scholar]
  • [6].Bursch W., Ellinger A., Gerner C., Fröhwein U., Schulte-Hermann R. Programmed cell death (PCD). Apoptosis, autophagic PCD, or others? Ann N Y Acad Sci. 2000;926:1–12. doi: 10.1111/j.1749-6632.2000.tb05594.x. [DOI] [PubMed] [Google Scholar]
  • [7].Rubinsztein D.C., DiFiglia M., Heintz N., Nixon R.A., Qin Z.H., Ravikumar B., et al. Autophagy and its possible roles in nervous system diseases, damage and repair. Autophagy. 2005;1:11–22. doi: 10.4161/auto.1.1.1513. [DOI] [PubMed] [Google Scholar]
  • [8].Shintani T., Klionsky D.J. Autophgy in health and disease: a doubleedged sword. Science. 2004;306:990–995. doi: 10.1126/science.1099993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [9].Diskin T., Tal-Or P., Erlich S., Mizrachy L., Alexandrovich A., Shohami E., et al. Closed head injury induces upregulation of Beclin 1 at the cortical site of injury. J Neurotrauma. 2005;22:750–762. doi: 10.1089/neu.2005.22.750. [DOI] [PubMed] [Google Scholar]
  • [10].Erlich S., Shohami E., Pinkas-Kramarski R. Neurodegeneration induces upregulation of Beclin 1. Autophagy. 2006;2:49–51. doi: 10.4161/auto.2156. [DOI] [PubMed] [Google Scholar]
  • [11].Erlich S., Alexandrovich A., Shohami E., Pinkas-Kramarski R. Rapamycin is a neuroprotective treatment for traumatic brain injury. Neurobiol Dis. 2007;26:86–93. doi: 10.1016/j.nbd.2006.12.003. [DOI] [PubMed] [Google Scholar]
  • [12].Clark R.S., Bayir H., Chu C.T., Alber S.M., Kochanek P.M., Watkins S.C. Autophagy is increased in mice after traumatic brain injury and is detectable in human brain after trauma and critical illness. Autophagy. 2008;4:88–90. doi: 10.4161/auto.5173. [DOI] [PubMed] [Google Scholar]
  • [13].Tao L.Y., Chen X.P., Ding M. The expression of caspase-3 after brain contusion in different severity in rat. J Forensic Med. 2003;19:4–7. [PubMed] [Google Scholar]
  • [14].Ashford T.P., Porter K.R. Cytoplasmic components in hepatic cell lysosomes. J Cell Biol. 1962;12:198–202. doi: 10.1083/jcb.12.1.198. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [15].Juhasz G., Neufeld T.P. Autophagy: a forty-year search for a missing membrane source. PLoS Biol. 2006;4:e36. doi: 10.1371/journal.pbio.0040036. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [16].Zhang Y.B., Chen X.P., Tao L.Y., Qin Z.H., Li S.X., Yang L., et al. Expression of cathepsin-B and-D after traumatic brain injury in rat. J Forensic Med. 2006;22:404–406. [PubMed] [Google Scholar]
  • [17].Sugawara K., Suzuki N.N., Fujioka Y., Mizushima N., Ohsumi Y., Inagaki F. The crystal structure of microtubule-associated protein light chain 3, a mammalian homologue of Saccharomyces cerevisiae Atg8. Gen Cell. 2004;9:611–618. doi: 10.1111/j.1356-9597.2004.00750.x. [DOI] [PubMed] [Google Scholar]
  • [18].Kouno T., Mizuguchi M., Tanida I., Ueno T., Kanematsu T., Mori Y., et al. Solution structure of microtubule-associated protein light chain 3 and identification of its functional subdomains. J Biol Chem. 2005;280:24610–24617. doi: 10.1074/jbc.M413565200. [DOI] [PubMed] [Google Scholar]
  • [19].Kabeya Y., Mizushima N., Yamamoto A., Oshitani-Okamoto S., Ohsumi Y., Yoshimori T. LC3, GABARAP and GATE16 localize to autophagosomal membrane depending on form-II formation. J Cell Sci. 2004;117(Pt13):2805–2812. doi: 10.1242/jcs.01131. [DOI] [PubMed] [Google Scholar]
  • [20].Asanuma K., Tanida I., Shirato I., Ueno T., Takahara H., Nishitani T., et al. MAP-LC3, a promising autophagosomal marker, is processed during the differentiation and recovery of podocytes from PAN nephrosis. FASEB J. 2003;17:1165–1167. doi: 10.1096/fj.02-0580fje. [DOI] [PubMed] [Google Scholar]
  • [21].Mizushima N. Methods for monitoring autophagy. Int J Biochem Cell Biol. 2004;36:2491–2502. doi: 10.1016/j.biocel.2004.02.005. [DOI] [PubMed] [Google Scholar]
  • [22].Tanida I., Ueno T., Kominami E. LC3 conjugation system in mammalian autophagy. Int J Biochem Cell Biol. 2004;36:2503–2518. doi: 10.1016/j.biocel.2004.05.009. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [23].Wu J.X., Dang Y.J., Su W., Liu C., Ma H.J., Shan Y.X., et al. Molecular cloning and characterization of rat LC3A and LC3B-Two novel markers of autophagosome. Biochem Biophys Res Commun. 2006;339:437–442. doi: 10.1016/j.bbrc.2005.10.211. [DOI] [PubMed] [Google Scholar]
  • [24].Kabeya Y., Mizushima N., Ueno T., Yamamoto A., Kirisako T., Noda T., et al. LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing. EMBO J. 2000;19:5720–5728. doi: 10.1093/emboj/19.21.5720. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [25].He H., Dang Y., Dai F., Guo Z., Wu J., She X., et al. Post-translational modifications of three members of the human MAP1LC3 family and detection of a novel type of modification for MAP1LC3B. J Biol Chem. 2003;278:29278–29287. doi: 10.1074/jbc.M303800200. [DOI] [PubMed] [Google Scholar]
  • [26].Tanida I., Ueno T., Kominami E. Human light chain 3/MAP1LC3B is cleaved at its carboxyl-terminal Met121 to expose Gly120 for lipidation and targeting to autophagosomal membranes. J Biol Chem. 2004;279:47704–47710. doi: 10.1074/jbc.M407016200. [DOI] [PubMed] [Google Scholar]
  • [27].Meijer A.J., Codogno P. Regulation and role of autophagy in mammalian cells. Int J Biochem Cell Biol. 2004;36:2445–2462. doi: 10.1016/j.biocel.2004.02.002. [DOI] [PubMed] [Google Scholar]
  • [28].Liang X.H., Kleeman L.K., Jiang H.H., Gordon G., Goldman J.E., Berry G., et al. Protection against fetal Sindbis virus encephalitis by Beclin, a novel Bcl-2 interacting protein. J Virol. 1998;72:8586–8596. doi: 10.1128/jvi.72.11.8586-8596.1998. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [29].Elmore S.P., Qian T., Grissom S.F., Lemasters J.J. The mitochondrial permeability transition initiates autophagy in rat hepatocytes. FASEB J. 2001;15:2286–2287. doi: 10.1096/fj.01-0206fje. [DOI] [PubMed] [Google Scholar]
  • [30].Bauvy C., Gane P., Arico S., Codogno P., Ogier-Denis E. Autophagy delays sulindac sulfide-induced apoptosis in the human intestinal colon cancer line HT-29. Exp Cell Res. 2001;268:139–149. doi: 10.1006/excr.2001.5285. [DOI] [PubMed] [Google Scholar]

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