Improvement in neuronal survival after ischemic preconditioning in hippocampal slice cultures

被引:69
作者
Xu, GP [1 ]
Dave, KR [1 ]
Vivero, R [1 ]
Schmidt-Kastner, R [1 ]
Sick, TJ [1 ]
Pérez-Pinzón, MA [1 ]
机构
[1] Univ Miami, Sch Med, Dept Neurol, Miami, FL 33101 USA
关键词
anoxia; ischemia; cell culture; tolerance;
D O I
10.1016/S0006-8993(02)02988-8
中图分类号
Q189 [神经科学];
学科分类号
071006 ;
摘要
The main goals of the current study were to assess: (a) whether a sublethal ischemic insult could protect the CA1 subregion of the hippocampus in organotypic slices against a lethal ischemic insult; and (b) whether this protection is long lasting as determined with an accurate immunohistochemical neuronal marker, NeuN. Hippocampal slice cultures were grown for 12-14 days in vitro. Slices were exposed either to oxygen/glucose deprivation (OGD) for 45 min (ischemia), or OGD for 15 min (ischemic preconditioning), 48 h prior to 45 min OGD, or were untreated (sham). Cell death was estimated by propidium iodide fluorescence I day after OGD and by NeuN immunohistochemistry 7 days after OGD. Image analysis was employed to measure the relative optical density of the NeuN-signal in all groups. After ischemia, damaged neurons were shrunken or lost and NeuN immmoreactivity was reduced. Relative optical density of NeuN (ROD [NeuN]) was 0.193 +/- 0.015 in control (sham) (n = 9). In slices that underwent ischemia, ROD [NeuN] declined to 0.108 +/- 0.018 (n = 5) in CA1 (*P < 0.05 ROD [NeuN] in preconditioned slice cultures was 0.190 +/- 0.037 (76% higher than the ischemia group). Similar results were found after measuring PI fluorescence. In the CA1 sub-region, PI fluorescence was about 13, 47 and 17% in the sham, ischemic and IPC groups, respectively. We suggest that the immmohistochemical approach validates the dye uptake method used in slice cultures and yields quantitative data specific for neurons. We also conclude that the organotypic hippocampal slice model is useful for studying delayed ischemic preconditioning that is maintained for hours or days after the preconditioning event. (C) 2002 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:153 / 158
页数:6
相关论文
共 26 条
  • [11] Comparison of excitotoxic profiles of ATPA, AMPA, KA and NMDA in organotypic hippocampal slice cultures
    Kristensen, BW
    Noraberg, J
    Zimmer, J
    [J]. BRAIN RESEARCH, 2001, 917 (01) : 21 - 44
  • [12] A simple in vitro model of ischemia based on hippocampal slice cultures and propidium iodide fluorescence
    Laake, JH
    Haug, FM
    Wieloch, T
    Ottersen, OP
    [J]. BRAIN RESEARCH PROTOCOLS, 1999, 4 (02): : 173 - 184
  • [13] MULLEN RJ, 1992, DEVELOPMENT, V116, P201
  • [14] PerezPinzon MA, 1996, NEUROSCIENCE, V75, P687
  • [15] Rapid preconditioning protects rats against ischemic neuronal damage after 3 but not 7 days of reperfusion following global cerebral ischemia
    PerezPinzon, MA
    Xu, GP
    Dietrich, WD
    Rosenthal, M
    Sick, TJ
    [J]. JOURNAL OF CEREBRAL BLOOD FLOW AND METABOLISM, 1997, 17 (02) : 175 - 182
  • [16] Ischaemic pre-conditioning in organotypic hippocampal slice cultures is inversely correlated to the induction of the 72 kDa heat shock protein (HSP72)
    Pringle, AK
    Thomas, SJ
    Signorelli, F
    Iannotti, F
    [J]. BRAIN RESEARCH, 1999, 845 (02) : 152 - 164
  • [17] Pringle AK, 2000, ACT NEUR S, V76, P79
  • [18] Preconditioning of primary rat neuronal cultures against ischemic injury: Characterization of the 'time window of protection'
    Reshef, A
    Sperling, O
    ZorefShani, E
    [J]. BRAIN RESEARCH, 1996, 741 (1-2) : 252 - 257
  • [19] ADAPTATION OF ADULT BRAIN-TISSUE TO ANOXIA AND HYPOXIA INVITRO
    SCHURR, A
    REID, KH
    TSENG, MT
    WEST, C
    RIGOR, BM
    [J]. BRAIN RESEARCH, 1986, 374 (02) : 244 - 248
  • [20] THE MECHANISM OF NEURONAL RESISTANCE AND ADAPTATION TO HYPOXIA
    SCHURR, A
    RIGOR, BM
    [J]. FEBS LETTERS, 1987, 224 (01): : 4 - 8