Clinical and Experimental Pediatrics

The Korean Pediatric Society (대한소아청소년과학회)
권호 표지

Fas/FasL expression in the hippocampus of neonatal rat brains follwing hypoxic-ischemic injury

저산소성 허혈성 손상을 받은 신생 흰쥐 뇌 해마에서 Fas와 FasL 단백 발현

  • Chang, Young Pyo (Department of Pediatrics, College of Medicine, Dankook University) ;
  • Kim, Myeung Ju (Department of Anatomy, College of Medicine, Dankook University) ;
  • Lee, Young Il (Department of Anatomy, College of Medicine, Dankook University) ;
  • Im, Ik Je (Department of Pediatrics, College of Medicine, Dankook University) ;
  • Cho, Jae Ju (Department of Pediatrics, College of Medicine, Dankook University) ;
  • Kim, Jong Wan (Department of Biological Science, Dankook University) ;
  • Yeo, Sung Moon (Department of Biological Science, Dankook University)
  • 장영표 (단국대학교 의과대학 소아과학교실) ;
  • 김명주 (단국대학교 의과대학 해부학교실) ;
  • 이영일 (단국대학교 의과대학 해부학교실) ;
  • 임익제 (단국대학교 의과대학 소아과학교실) ;
  • 조재주 (단국대학교 의과대학 소아과학교실) ;
  • 김종완 (첨단과학대학 생물학과) ;
  • 여성문 (첨단과학대학 생물학과)
  • Received : 2005.09.14
  • Accepted : 2005.10.24
  • Published : 2006.02.15
Download PDF
⟨ Previous Next ⟩

Abstract

Purpose : Fas is a cell surface receptor that transduces apoptotic death signals. Interaction of extracelluar domain of Fas with Fas ligand(FasL) triggers the apoptotic process in many diseases. We investigated the expression of Fas and FasL in the hippocampus of 7-day-old newborn rat brains following hypoxia-ischemia injury. Methods : The 7-days-old newborn rats were exposed to 8 percent oxygen for two hours after the ligation of right common carotid arteries. The newborn rats were killed and their brains were removed at 12, 14 and 48 hours after hypoxic-ischemic injury. The expressions of Fas and FasL of the right hippocampus were observed by western blotting and immunofluorescent staining. Results : Fas and FasL were strongly expressed in the right hippocampus ipsilateral to the ligation of the common carotid artery by western blotting at 12 hours following hypoxic-ischemic injury, and then slowly decreased. The immunofluorescent expressions of Fas and FasL strongly increased in the CA1 area of the right hippocampus at 12 and 24 hours following hypoxic-ischemic injury. The immunofluorescent expression of Fas decreased at 48 hours, but the expression of FasL persisted strongly at 48 hours following hypoxic-ischemic injury. Conclusion : The interaction of Fas with FasL on the cell surface may be involved in neuronal injury following hypoxic-ischemic injury in the developing brain.

목 적 : Fas는 세포표면 수용체로 세포사멸 신호를 전도한다. 많은 질환에서 세포표면의 Fas가 Fas ligand(FasL)와 결합하여 세포사멸 과정을 유발하게 된다. 연구자들은 7일된 신생 흰쥐에 저산소성 허혈성 손상을 유발한 후 뇌 해마에서 Fas와 FasL의 발현을 관찰하고자 하였다. 방 법 : 7일된 신생 흰쥐를 오른쪽 총 경동맥 영구 결찰 후 8% 산소에 2시간 노출시켰다. 저산소성 허혈성 손상 후 12, 24, 48시간에 뇌를 적출 냉동 보관하였다. Western blotting 방법과 면역형광염색 방법으로 냉동 보관된 뇌의 경동맥을 결찰한 오른 쪽 해마에서 Fas와 FasL의 발현을 관찰하였다. 결 과 : Fas와 FasL의 발현은 저산소성 허혈성 손상 후 12시간에 경동맥이 결찰된 오른쪽 해마에서 크게 증가하고 이후 감소하는 것을 western blotting 방법에 의해 관찰하였다. Fas와 FasL의 면역형광발현은 오른쪽 해마의 CA1 영역에서 손상 후 12시간과 24시간에 대조군에 비해 증가하였다. Fas의 면역형광 발현은 손상 후 48시간에 감소하였으나 FasL의 면역형광발현은 손상 후 48시간에도 지속되었다. 결 론 : 세포표면에서 Fas와 FasL의 발현과 그들의 결합은 저산소성 허혈성 손상을 받은 미성숙 뇌의 신경세포 손상에 기여할 것으로 추측되었다.

Keywords

Acknowledgement

Supported by : 단국대학교

References

  1. Vannucci RC. Hypoxic-ischemic encephalopathy. Am J Perinatol 2000;17:113-20 https://doi.org/10.1055/s-2000-9293
  2. Chang YP. Cellular and biochemical mechanism of perinatal hypoxic-ischemic brain injury. J Korean Pediatr Soc 2002; 45:560-7
  3. Dell'Anna E, Chen Y, Engidawork E, Andersson K, Lubec G, Luthman J, et al. Delayed neuronal death following perinatal asphyxia in rat. Exp Brain Res 1997;115:105-15 https://doi.org/10.1007/PL00005670
  4. Yue X, Mehmet H, Penrice J, Cooper C, Cady E, Wyatt JS, et al. Apoptosis and necrosis in the newborn piglet brain following transient cerebral hypoxia-ischaemia. Neuropathol Appl Neurobiol 1997;23:16-25 https://doi.org/10.1111/j.1365-2990.1997.tb01181.x
  5. Yoon BH, Romero R, Kim CJ, Koo JN, Choe G, Syn HC, et al. High expression of tumor necrosis factor-$\alpha$ and interleukin- 6 in periventricular leukomalacia. Am J Obstet Gynecol 1997;177:406-11 https://doi.org/10.1016/S0002-9378(97)70206-0
  6. Chang YP, Shin DH, Lee E, Kim JW, Kwon BS, Jung MK, et al. Protective effect of growth hormone on neuronal apoptosis after hypoxia-ischemia in the neonatal rat brain. Neurosci Lett 2004;354:64-8 https://doi.org/10.1016/j.neulet.2003.09.070
  7. Eischen CM, Leibson PJ. The Fas pathway in apoptosis. In : Kaufmann SH, editor. Apotosis. London; Academic Press, 1997:107-32 https://doi.org/10.1016/S1054-3589(08)61056-X
  8. Park C, Sakamaki K, Tachibana O, Yamashima T, Yamashita J, Yonehara S. Expression of fas antigen in the normal mouse brain. Biochem Biophys Res Commun 1998;252: 623-8 https://doi.org/10.1006/bbrc.1998.9572
  9. Bechmann I, Mor G, Nilsen J, Eliza M, Nitsch R, Naftolin F. FasL(CD95L, Apo1L) is expressed in the normal rat and human brain : evidence for the existence of an immunological brain barrier. Glia 1999;27:62-74 https://doi.org/10.1002/(SICI)1098-1136(199907)27:1<62::AID-GLIA7>3.0.CO;2-S
  10. Choi C, Park JY, Lee J, Lim JH, Shin EC, Ahn YS, et al. Fas ligand and Fas are expressed constitutively in human astrocytes and the expression increases with IL-1, IL-6, TNF-alpha, or IFN-gamma. J Immunol 1999;162:1889-95
  11. Dowling P, Shang G, Raval S, Menonna J, Cook S, Husar W. Involvement of the CD95(APO-1/Fas) receptor/ligand system in multiple sclerosis brain. J Exp Med 1996;184: 1513-8 https://doi.org/10.1084/jem.184.4.1513
  12. Ciusani E, Frigerio S, Gelati M, Corsini E, Dufour A, Nespolo A, et al. Soluble Fas(Apo-1) levels in cerebrospinal fluid of multiple sclerosis patients. J Neuroimmunol 1998; 82:5-12 https://doi.org/10.1016/S0165-5728(97)00177-X
  13. Nishimura T, Akiyama H, Yonehara S, Kondo H, Ikeda K, Kato M, et al. Fas antigen expression in brains of patients with Alzheimer-type dementia. Brain Res 1995;695:137-45 https://doi.org/10.1016/0006-8993(95)00699-Q
  14. Guo Q, Sopher BL, Furukawa K, Pham DG, Robinson N, Martin GM, et al. Alzheimer's presenilin mutation sensitizes neural cells to apoptosis induced by trophic factor withdrawal and amyloid beta-peptide : involvement of calcium and oxyradicals. J Neurosci 1997;17:4212-22 https://doi.org/10.1523/JNEUROSCI.17-11-04212.1997
  15. Mogi M, Harada M, Kondo T, Mizuno Y, Narabayashi H, Riederer P, et al. The soluble form of Fas molecule is elevated in parkinsonian brain tissues. Neurosci Lett 1996;220: 195-8 https://doi.org/10.1016/S0304-3940(96)13257-2
  16. Vogt M, Bauer MK, Ferrari D, Schulze-Osthoff K. Oxidative stress and hypoxia/reoxygenation trigger CD95(APO- 1/Fas) ligand expression in microglial cells. FEBS Lett 1998;429:67-72 https://doi.org/10.1016/S0014-5793(98)00562-6
  17. Martin-Villalba A, Herr I, Jeremias I, Hahne M, Brandt R, Vogel J, et al. CD95 ligand(Fas-L/APO-1L) and tumor necrosis factor-related apoptosis-inducing ligand mediate ischemia- induced apoptosis in neurons. J Neurosci 1999;19: 3809-17 https://doi.org/10.1523/JNEUROSCI.19-10-03809.1999
  18. Li GL, Farooque M, Olsson Y. Changes of Fas and Fas ligand immunoreactivity after compression trauma to rat spinal cord. Acta Neuropathol(Berl) 2000;100:75-81 https://doi.org/10.1007/s004010051195
  19. Rosenbaum DM, Gupta G, D'Amore J, Singh M, Weidenheim K, Zhang H, et al. Fas(CD95/APO-1) plays a role in the pathophysiology of focal cerebral ischemia. J Neurosci Res 2000;61:686-92 https://doi.org/10.1002/1097-4547(20000915)61:6<686::AID-JNR12>3.0.CO;2-7
  20. Bechmann I, Lossau S, Steiner B, Mor G, Gimsa U, Nitsch R. Reactive astrocytes upregulate Fas(CD95) and Fas ligand(CD95L) expression but do not undergo programmed cell death during the course of anterograde degeneration. Glia 2000;32:25-41 https://doi.org/10.1002/1098-1136(200010)32:1<25::AID-GLIA30>3.0.CO;2-Y
  21. Jin K, Graham SH, Mao X, Nagayama T, Simon RP, Greenberg DA. Fas(CD95) may mediate delayed cell death in hippocampal CA1 sector after global cerebral ischemia. J Cereb Blood Flow Metab 2001;21:1411-21 https://doi.org/10.1097/00004647-200112000-00005
  22. Tan Z, Levid J, Schreiber SS. Increased expression of Fas (CD95/APO-1) in adult rat brain after kainate-induced seizures. Neuroreport 2001;12:1979-82 https://doi.org/10.1097/00001756-200107030-00040
  23. Padosch SA, Popp E, Vogel P, Bottiger BW. Altered protein expression levels of Fas/CD95 and Fas ligand in differentially vulnerable brain areas in rats after global cerebral ischemia. Neurosci Lett 2003;338:247-51 https://doi.org/10.1016/S0304-3940(02)01408-8
  24. Felderhoff-Mueser U, Taylor DL, Greenwood K, Kozma M, Stibenz D, Joashi UC, et al. Fas/CD95/APO-1 can function as a death receptor for neuronal cells in vitro and in vivo and is upregulated following cerebral hypoxic-ischemic injury to the developing rat brain. Brain Pathol 2000;10:17- 29 https://doi.org/10.1111/j.1750-3639.2000.tb00239.x
  25. Northington FJ, Ferriero DM, Flock DL, Martin LJ. Delayed neurodegeneration in neonatal rat thalamus after hypoxia-ischemia is apoptosis. J Neurosci 2001;21:1931-8 https://doi.org/10.1523/JNEUROSCI.21-06-01931.2001
  26. van Landeghem FK, Felderhoff-Mueser U, Moysich A, Stadelmann C, Obladen M, Bruck W, et al. Fas(CD95/Apo- 1)/Fas ligand expression in neonates with pontosubicular neuron necrosis. Pediatr Res 2002;51:129-35 https://doi.org/10.1203/00006450-200202000-00003