Improvement in neuronal survival after ischemic preconditioning in hippocampal slice cultures

Xu, Guang Ping; Dave, Kunjan R.; Vivero, Richard J.; Schmidt‐Kastner, Rainald; Sick, Thomas J.; Pérez-Pinzón, Miguel A.

doi:10.1016/s0006-8993(02)02988-8

Cited by 73 publications

(73 citation statements)

References 22 publications

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“…They observed that the lesions induced by OGD were similar to those shown by animals submitted to transient cerebral ischemia. We corroborated recently that, like global cerebral ischemia, OGD promotes selective cell death in the CA1 subregion of the hippocampus (Xu et al, 2002). The slices were washed three times with glucose-free HBSS, pH 7.4, containing the following (in mM): 1.26 CaCl 2 ⅐ 2 H 2 O, 5.37 KCl, 0.44 KH 2 PO 4 , 0.49 MgCl 2 , 0.41 MgSO 4 ⅐ 7 H 2 O, 136.9 NaCl, 4.17 NaHCO 3 , 0.34 Na 2 HPO 4 ⅐ 7 H 2 O, and 15 sucrose (all from Sigma).…”

Section: Oxygen-glucose Deprivationsupporting

confidence: 83%

“…We defined the ischemia and preconditioning protocols in a previous study (Xu et al, 2002). The organotypic cultures have been used to study mechanisms underlying neuronal death induced by hypoxia-aglycemia (Pringle et al, 1997a) and excitotoxins (Sakaguchi et al, 1997).…”

Section: Oxygen-glucose Deprivationmentioning

confidence: 99%

“…We reported recently that sublethal in vitro ischemia in organotypic hippocampal slice cultures protects against neuronal cell death produced by lethal in vitro ischemia (Xu et al, 2002). The present study, using the organotypic slice cultures, investigates three issues concerning the mechanism of IPC: (1) whether the NMDA receptors are involved in the triggering phase of IPC via calcium, (2) whether the PKC isozymes are involved in induction of neuroprotection, and (3) whether ⑀PKC is involved in the signaling pathway of IPC neuroprotection as shown in the heart (Souroujon and Mochly-Rosen, 1998).…”

Section: Introductionmentioning

confidence: 99%

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εPKC Is Required for the Induction of Tolerance by Ischemic and NMDA-Mediated Preconditioning in the Organotypic Hippocampal Slice

et al. 2003

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Glutamate receptors and calcium have been implicated as triggering factors in the induction of tolerance by ischemic preconditioning (IPC) in the brain. However, little is known about the signal transduction pathway that ensues after the IPC induction pathway. The main goals of the present study were to determine whether NMDA induces preconditioning via a calcium pathway and promotes translocation of the protein kinase C ⑀ (⑀PKC) isozyme and whether this PKC isozyme is key in the IPC signal transduction pathway. We corroborate here that IPC and a sublethal dose of NMDA were neuroprotective, whereas blockade of NMDA receptors during IPC diminished IPC-induced neuroprotection. Calcium chelation blocked the protection afforded by both NMDA and ischemic preconditioning significantly, suggesting a significant role of calcium. Pharmacological preconditioning with the nonselective PKC isozyme activator phorbol myristate acetate could not emulate IPC, but blockade of PKC activation with chelerythrine during IPC blocked its neuroprotection. These results suggested that there might be a dual involvement of PKC isozymes during IPC. This was corroborated when neuroprotection was blocked when we inhibited ⑀PKC during IPC and NMDA preconditioning, and IPC neuroprotection was emulated with the activator of ⑀PKC. The possible correlation between NMDA, Ca 2ϩ , and ⑀PKC was found when we emulated IPC with the diacylglycerol analog oleoylacetyl glycerol, suggesting an indirect pathway by which Ca 2ϩ could activate the calcium-insensitive ⑀PKC isozyme. These results demonstrated that the ⑀PKC isozyme played a key role in both IPC-and NMDA-induced tolerance.

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Section: Oxygen-glucose Deprivationsupporting

confidence: 83%

Section: Oxygen-glucose Deprivationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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εPKC Is Required for the Induction of Tolerance by Ischemic and NMDA-Mediated Preconditioning in the Organotypic Hippocampal Slice

et al. 2003

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“…Animals were decapitated and the brains quickly removed. Organotypic hippocampal slice cultures were prepared as described previously (Xu et al, 2002). In brief, transverse slices (400 μm) were dissected from the hippocampi and placed in Gey's Balanced Salt Solution (all chemicals were obtained from Sigma St. Louis, MO, or specified otherwise) supplemented with 6.5 mg/ml glucose at 4°C.…”

Section: Preparation Of Organotypic Slice Culturesmentioning

confidence: 99%

ɛPKC phosphorylates the mitochondrial K+ATP channel during induction of ischemic preconditioning in the rat hippocampus

et al. 2007

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Neuroprotection against cerebral ischemia conferred by ischemic preconditioning (IPC) requires translocation of epsilon protein kinase C (εPKC). A major goal in our laboratory is to define the cellular targets by which εPKC confers protection. We tested the hypothesis that εPKC targets the mitochondrial channel ( ) after IPC. Our results demonstrated a rapid translocation of εPKC to rat hippocampal mitochondria after IPC. Because in other tissues εPKC targets channels, but its presence in brain mitochondria is controversial, we determined the presence of the channel-specific subunits (Kir6.1 and Kir6.2) in mitochondria isolated from rat hippocampus. Next, we determined whether channels play a role in the IPC induction. In hippocampal organotypic slice cultures, IPC and lethal ischemia were induced by oxygen-glucose deprivation. Subsequent cell death in the CA1 region was quantified using propidium iodide staining. Treatment with the channel openers diazoxide or pinacidil 48 h prior to lethal ischemia protected hippocampal CA1 neurons, mimicking the induction of neuroprotection conferred by either IPC or εPKC agonist-induced preconditioning. Blockade of channels using 5-hydroxydecanoic acid abolished the neuroprotection due to either IPC or εPKC preconditioning. Both ischemic andεPKC agonist-mediated preconditioning resulted in phosphorylation of the channel subunit Kir6.2. After IPC, selective inhibition of εPKC activation prevented Kir6.2 phosphorylation, consistent with Kir6.2 as a phosphorylation target of εPKC or its downstream effectors. Our results support the hypothesis that the brain channel is an important target of IPC and the signal transduction pathways initiated by εPKC.

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“…Our ischemia protocols have been described in previous studies (Xu et al, 2002;Raval et al, 2003). For OGD, slices were washed three times with aglycemic Hank's balanced salt solution (AHBSS) (pH 7.4) of the following constitution: CaCl 2 Á 2H 2 O 1.26 mmol/l, KCl 5.37 mmol/L, KH 2 PO 4 0.44 mmol/L, MgCl 2 0.49 mmol/L, MgSO 4 Á 7H 2 O 0.41 mmol/L, NaCl 136.9 mmol/L, NaHCO 3 4.17 mmol/L, Na 2 HPO 4 Á 7H 2 O 0.34 mmol/L, sucrose 15 mmol/L (all Sigma, St Louis, MO, USA).…”

Section: Induction Of Ischemia (Oxygen/glucose Deprivation)mentioning

confidence: 99%

Protein Kinase C Delta Cleavage Initiates an Aberrant Signal Transduction Pathway after Cardiac Arrest and Oxygen Glucose Deprivation

Raval

Dave

Prado

et al. 2005

J Cereb Blood Flow Metab

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Protein kinase C (PKC) isozymes have been known to mediate a variety of complex and diverse cellular functions. dPKC has been implicated in mediating apoptosis. Using two models of cerebral ischemia, cardiac arrest in rats and oxygen glucose deprivation (OGD) in organotypic hippocampal slices, we tested whether an ischemic insult promoted dPKC cleavage during the reperfusion and whether the upstream pathway involved release of cytochrome c and caspase 3 cleavage. We showed that cardiac arrest/OGD significantly enhanced dPKC translocation and increased its cleavage at 3 h of reperfusion. Since dPKC is one of the substrates for caspase 3, we next determined caspase 3 activation after cardiac arrest and OGD. The maximum decrease in levels of procaspase 3 was observed at 3 h of reperfusion after cardiac arrest and OGD. We also determined cytochrome c release, since it is upstream of caspase 3 activation. Cytochrome c in cytosol increased at 1 h of reperfusion after cardiac arrest/OGD. Inhibition of either dPKC/caspase 3 during OGD and early reperfusion resulted in neuroprotection in CA1 region of hippocampus. Our results support the deleterious role of dPKC in reperfusion injury. We propose that early cytochrome c release and caspase 3 activation promote dPKC translocation/cleavage.

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Improvement in neuronal survival after ischemic preconditioning in hippocampal slice cultures

Cited by 73 publications

References 22 publications

εPKC Is Required for the Induction of Tolerance by Ischemic and NMDA-Mediated Preconditioning in the Organotypic Hippocampal Slice

εPKC Is Required for the Induction of Tolerance by Ischemic and NMDA-Mediated Preconditioning in the Organotypic Hippocampal Slice

ɛPKC phosphorylates the mitochondrial K+ATP channel during induction of ischemic preconditioning in the rat hippocampus

Protein Kinase C Delta Cleavage Initiates an Aberrant Signal Transduction Pathway after Cardiac Arrest and Oxygen Glucose Deprivation

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