Mechanisms and Effects of Intracellular Calcium Buffering on Neuronal Survival in Organotypic Hippocampal Cultures Exposed to Anoxia/Aglycemia or to Excitotoxins

Abdel-Hamid, Khaled; Tymianski, Michael

doi:10.1523/jneurosci.17-10-03538.1997

Cited by 117 publications

(49 citation statements)

References 96 publications

(151 reference statements)

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“…Intensity of PI fluorescence has been correlated with other markers of neuronal loss and has been shown to provide a quantitative index of cell death (Abdel-Hamid and Tymianski, 1997). In order to compare different treatment groups, PI uptake is often expressed as a percentage of maximal uptake.…”

Section: Discussionmentioning

confidence: 99%

A method for objectively quantifying propidium iodide exclusion in organotypic hippocampal slice cultures

Happ

Tasker

2016

Journal of Neuroscience Methods

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Section: Discussionmentioning

confidence: 99%

A method for objectively quantifying propidium iodide exclusion in organotypic hippocampal slice cultures

Happ

Tasker

2016

Journal of Neuroscience Methods

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“…The onset of cellular death observed in this in vitro model happens earlier than death in in vivo models (cell death in vivo occurs 3 or 4 days after injury) (4), probably because the culture condition by itself stress the cells, and other signaling cascades than those acting in vivo could be activated. Despite that different characteristics, the organotypic culture of hippocampus is a well accepted model and is widely used to mimic ischemic condition in vitro (2,(8)(9)(10)(11)17,18).…”

Section: Discussionmentioning

confidence: 99%

“…Many aspects of ischemic cell death have been demonstrated in animal models (6,3), however, these animal models reveal a complex way to dissect the cellular and molecular mechanisms involved in ischemic neurodegeneration. Organotypic slice cultures of hippocampal tissue have become an attractive alternative to study neuronal death induced by treatments such as hypoxia (7), hypoglycemia (8), excitotoxins (9) and oxygen and glucose deprivation (2,10). This in vitro technique is a very useful tool for studying the physiological and pharmacological properties of neuronal circuits with the major advantage that it is a well preserved organotypic organization of the tissue (11).…”

Section: Introductionmentioning

confidence: 99%

Cellular Death in Hippocampus in Response to PI3K Pathway Inhibition and Oxygen and Glucose Deprivation

et al. 2005

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We investigated the importance of the phosphoinositide3-kinase (PI3K) pathway in CA1 and dentate gyrus (DG) areas of hippocampus by exposing organotypic cultures to LY294002, a PI3K inhibitor, or to oxygen and glucose deprivation (OGD) for up to 21 hours. LY294002 induced increased propidium iodide (PI) uptake and caspase 3/7 activity in both regions, with a faster onset in DG. In contrast, cultures exposed to 60 min of OGD showed a PI uptake only in the CA1 area, beginning 13 h after the insult and increasing until 21 h. We did not observe any significant changes in AKT phosphorylation and immunocontent in CA1 or DG areas of organotypic cultures exposed to OGD, suggesting that the phosphorylation of this protein at Ser-473 is unrelated to the cellular damage induced by ischemia. Our results suggest that the inhibition of the PI3K pathway does not mimic the cell death profile observed with an ischemic model.

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“…These conditions, which are required for the activation of PT, develop in neuronal cell cultures subjected to oxidative stress, excitotoxicity, and oxygen-glucose deprivation. In these settings, the sequestration of high levels of calcium by mitochondria is suggested to be the underlying cause of cell death (21,22). In vivo parameters required for the induction of PT are mimicked during ischemia͞reperfusion in the brain.…”

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confidence: 99%

Cyclophilin D is a component of mitochondrial permeability transition and mediates neuronal cell death after focal cerebral ischemia

Schinzel

Takeuchi²,

Huang³

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

756

650

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Mitochondrial permeability transition (PT) is a phenomenon induced by high levels of matrix calcium and is characterized by the opening of the PT pore (PTP). Activation of the PTP results in loss of mitochondrial membrane potential, expansion of the matrix, and rupture of the mitochondrial outer membrane. Consequently, PT has been implicated in both apoptotic and necrotic cell death. Cyclophilin D (CypD) appears to be a critical component of the PTP. To investigate the role of CypD in cell death, we created a CypD-deficient mouse. In vitro, CypD-deficient mitochondria showed an increased capacity to retain calcium and were no longer susceptible to PT induced by the addition of calcium. CypD-deficient primary mouse embryonic fibroblasts (MEFs) were as susceptible to classical apoptotic stimuli as the WT, suggesting that CypD is not a central component of cell death in response to these specific death stimuli. However, CypD-deficient MEFs were significantly less susceptible than their WT counterparts to cell death induced by hydrogen peroxide, implicating CypD in oxidative stress-induced cell death. Importantly, CypD-deficient mice displayed a dramatic reduction in brain infarct size after acute middle cerebral artery occlusion and reperfusion, strongly supporting an essential role for CypD in an ischemic injury model in which calcium overload and oxidative stress have been implicated. mitochondria ͉ oxidative stress ͉ calcium homeostasis N umerous toxic stimuli convert mitochondria from a lifesustaining organelle to an inducer of cell death (1-3). In response to cell death stimuli, prodeath proteins sequestered in the inner mitochondrial membrane space are released into the cytosol upon disruption of the mitochondrial outer membrane (MOM) (4-6). The proapoptotic BCL-2 family members BAX and BAK constitute a ''gateway'' to the apoptotic program by regulating events leading to disruption of the MOM (7). BCL-2 family members not only function at the MOM but also are located at the endoplasmic reticulum (ER), where they regulate calcium fluxes. By controlling steady-state calcium levels in the ER, BCL-2 family members regulate the amount of calcium released from the ER to transmit a death signal mediated by mitochondrial calcium uptake (8-11).Sequestration of high levels of calcium by mitochondria can also lead to disruption of the MOM, which, mechanistically, is thought to occur by means of the phenomenon of permeability transition (PT). PT is described as an abrupt increase of inner membrane permeability to solutes with molecular masses of Ͻ1,500 Da. These events are caused by the opening of a highly regulated channel [PT pore (PTP)], which leads to dissipation of the mitochondrial transmembrane potential and an influx of solutes, causing expansion of the matrix (reviewed in ref. 12). The latter event may result in sufficient swelling to rupture the MOM and cause cytochrome c release and subsequent caspase activation, resulting in apoptosis. However, dissipation of the membrane potential can also lead to a sudden d...

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Mechanisms and Effects of Intracellular Calcium Buffering on Neuronal Survival in Organotypic Hippocampal Cultures Exposed to Anoxia/Aglycemia or to Excitotoxins

Cited by 117 publications

References 96 publications

A method for objectively quantifying propidium iodide exclusion in organotypic hippocampal slice cultures

A method for objectively quantifying propidium iodide exclusion in organotypic hippocampal slice cultures

Cellular Death in Hippocampus in Response to PI3K Pathway Inhibition and Oxygen and Glucose Deprivation

Cyclophilin D is a component of mitochondrial permeability transition and mediates neuronal cell death after focal cerebral ischemia

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