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Cognitive impairments accompanying rodent mild traumatic brain injury involve p53-dependent neuronal cell death and are ameliorated by the tetrahydrobenzothiazole PFT-α.

Rachmany L, Tweedie D, Rubovitch V, Yu QS, Li Y, Wang JY, Pick CG, Greig NH - PLoS ONE (2013)

Bottom Line: With parallels to concussive mild traumatic brain injury (mTBI) occurring in humans, anesthetized mice subjected to a single 30 g weight drop mTBI event to the right parietal cortex exhibited significant diffuse neuronal degeneration that was accompanied by delayed impairments in recognition and spatial memory.Cellular studies on human SH-SY5Y cells and rat primary neurons challenged with glutamate excitotoxicity and H2O2 induced oxidative stress, confirmed the ability of PFT-α and a close analog to protect against these TBI associated mechanisms mediating neuronal loss.These studies suggest that p53-dependent apoptotic mechanisms underpin the neuronal and cognitive losses accompanying mTBI, and that these are potentially reversible by p53 inactivation.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy and Anthropology, Sackler School of Medicine and Sagol School of Neuroscience, Tel Aviv University, Tel Aviv, Israel.

ABSTRACT
With parallels to concussive mild traumatic brain injury (mTBI) occurring in humans, anesthetized mice subjected to a single 30 g weight drop mTBI event to the right parietal cortex exhibited significant diffuse neuronal degeneration that was accompanied by delayed impairments in recognition and spatial memory. To elucidate the involvement of reversible p53-dependent apoptosis in this neuronal loss and associated cognitive deficits, mice were subjected to experimental mTBI followed by the systemic administration of the tetrahydrobenzothiazole p53 inactivator, PFT-α, or vehicle. Neuronal loss was quantified immunohistochemically at 72 hr. post-injury by the use of fluoro-Jade B and NeuN within the dentate gyrus on both sides of the brain, and recognition and spatial memory were assessed by novel object recognition and Y-maze paradigms at 7 and 30 days post injury. Systemic administration of a single dose of PFT-α 1 hr. post-injury significantly ameliorated both neuronal cell death and cognitive impairments, which were no different from sham control animals. Cellular studies on human SH-SY5Y cells and rat primary neurons challenged with glutamate excitotoxicity and H2O2 induced oxidative stress, confirmed the ability of PFT-α and a close analog to protect against these TBI associated mechanisms mediating neuronal loss. These studies suggest that p53-dependent apoptotic mechanisms underpin the neuronal and cognitive losses accompanying mTBI, and that these are potentially reversible by p53 inactivation.

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p53 inhibition by PFT-α/analog inhibits glutamate-induced excitotoxicity and oxidative stress mediated loss of cell viability in neuronal cultures.Human SH-SY5Y cells were subjected to p53 inactivation (PFT-α analog 1 to 10 μM) and then challenged with (A) glutamate (100 mM) excitotoxicity or (B) oxidative stress (H2O2: 500 μM). These insults alone significantly reduced cellular viability (* p<0.05 vs. control, Dunnetts t-test), which was mitigated by p53 inactivation (# p<0.05 vs. glutamate alone, Dunnetts t-test). (C) Rat primary cortical neuron cultures undergo time-dependent degeneration [44] that was mitigated by the addition of PFT-α (2 nM to 1 μM; * p<0.05, ** p<0.01, *** p<0.001 vs. untreated controls that are expressed as 100% (Dunnett’s t-test). A 10 μM PFT-α concentration proved to be toxic to primary neurons (*** p<0.001 vs. untreated controls; Dunnett’s t-test). (D) In an alike manner to SH-SY5Y cells, exposure of primary cortical neurons to glutamate (100 μM) resulted in reduced survival (* p<0.05 vs. control, Dunnetts t-test),) and pre-treatment with 2 to 100 nM PFT-α ameliorated this (NS not significantly different from untreated controls, Dunnetts t-test). Analysis of viable neurons was undertaken by MTS assay at 24 hr.
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pone-0079837-g002: p53 inhibition by PFT-α/analog inhibits glutamate-induced excitotoxicity and oxidative stress mediated loss of cell viability in neuronal cultures.Human SH-SY5Y cells were subjected to p53 inactivation (PFT-α analog 1 to 10 μM) and then challenged with (A) glutamate (100 mM) excitotoxicity or (B) oxidative stress (H2O2: 500 μM). These insults alone significantly reduced cellular viability (* p<0.05 vs. control, Dunnetts t-test), which was mitigated by p53 inactivation (# p<0.05 vs. glutamate alone, Dunnetts t-test). (C) Rat primary cortical neuron cultures undergo time-dependent degeneration [44] that was mitigated by the addition of PFT-α (2 nM to 1 μM; * p<0.05, ** p<0.01, *** p<0.001 vs. untreated controls that are expressed as 100% (Dunnett’s t-test). A 10 μM PFT-α concentration proved to be toxic to primary neurons (*** p<0.001 vs. untreated controls; Dunnett’s t-test). (D) In an alike manner to SH-SY5Y cells, exposure of primary cortical neurons to glutamate (100 μM) resulted in reduced survival (* p<0.05 vs. control, Dunnetts t-test),) and pre-treatment with 2 to 100 nM PFT-α ameliorated this (NS not significantly different from untreated controls, Dunnetts t-test). Analysis of viable neurons was undertaken by MTS assay at 24 hr.

Mentions: As glutamate-induced excitotoxicity and oxidative stress together with natural degeneration [44], are considered to underpin, in part, the cellular loss in brain after a TBI insult [19,23,29], neuronal cultures were pretreated with vehicle or PFT-α/analog (primary cortical cells: 0, 2 nM to 10 μM PFT-α; human SH-SY5Y cells 1 to 10 μM PFT-α analog Y-6-159) and, 1 hr. later were exposed to glutamate (SH-SY5Y cells: 100 mM; primary cells: 100 μM, for 24 hr.) or oxidative stress (H2O2: 500 μM, 24 hr.) or no insult for naturally degenerating primary cortical cultures. The percent of neuronal survivals at 24 hr. are shown in Figure 2.


Cognitive impairments accompanying rodent mild traumatic brain injury involve p53-dependent neuronal cell death and are ameliorated by the tetrahydrobenzothiazole PFT-α.

Rachmany L, Tweedie D, Rubovitch V, Yu QS, Li Y, Wang JY, Pick CG, Greig NH - PLoS ONE (2013)

p53 inhibition by PFT-α/analog inhibits glutamate-induced excitotoxicity and oxidative stress mediated loss of cell viability in neuronal cultures.Human SH-SY5Y cells were subjected to p53 inactivation (PFT-α analog 1 to 10 μM) and then challenged with (A) glutamate (100 mM) excitotoxicity or (B) oxidative stress (H2O2: 500 μM). These insults alone significantly reduced cellular viability (* p<0.05 vs. control, Dunnetts t-test), which was mitigated by p53 inactivation (# p<0.05 vs. glutamate alone, Dunnetts t-test). (C) Rat primary cortical neuron cultures undergo time-dependent degeneration [44] that was mitigated by the addition of PFT-α (2 nM to 1 μM; * p<0.05, ** p<0.01, *** p<0.001 vs. untreated controls that are expressed as 100% (Dunnett’s t-test). A 10 μM PFT-α concentration proved to be toxic to primary neurons (*** p<0.001 vs. untreated controls; Dunnett’s t-test). (D) In an alike manner to SH-SY5Y cells, exposure of primary cortical neurons to glutamate (100 μM) resulted in reduced survival (* p<0.05 vs. control, Dunnetts t-test),) and pre-treatment with 2 to 100 nM PFT-α ameliorated this (NS not significantly different from untreated controls, Dunnetts t-test). Analysis of viable neurons was undertaken by MTS assay at 24 hr.
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pone-0079837-g002: p53 inhibition by PFT-α/analog inhibits glutamate-induced excitotoxicity and oxidative stress mediated loss of cell viability in neuronal cultures.Human SH-SY5Y cells were subjected to p53 inactivation (PFT-α analog 1 to 10 μM) and then challenged with (A) glutamate (100 mM) excitotoxicity or (B) oxidative stress (H2O2: 500 μM). These insults alone significantly reduced cellular viability (* p<0.05 vs. control, Dunnetts t-test), which was mitigated by p53 inactivation (# p<0.05 vs. glutamate alone, Dunnetts t-test). (C) Rat primary cortical neuron cultures undergo time-dependent degeneration [44] that was mitigated by the addition of PFT-α (2 nM to 1 μM; * p<0.05, ** p<0.01, *** p<0.001 vs. untreated controls that are expressed as 100% (Dunnett’s t-test). A 10 μM PFT-α concentration proved to be toxic to primary neurons (*** p<0.001 vs. untreated controls; Dunnett’s t-test). (D) In an alike manner to SH-SY5Y cells, exposure of primary cortical neurons to glutamate (100 μM) resulted in reduced survival (* p<0.05 vs. control, Dunnetts t-test),) and pre-treatment with 2 to 100 nM PFT-α ameliorated this (NS not significantly different from untreated controls, Dunnetts t-test). Analysis of viable neurons was undertaken by MTS assay at 24 hr.
Mentions: As glutamate-induced excitotoxicity and oxidative stress together with natural degeneration [44], are considered to underpin, in part, the cellular loss in brain after a TBI insult [19,23,29], neuronal cultures were pretreated with vehicle or PFT-α/analog (primary cortical cells: 0, 2 nM to 10 μM PFT-α; human SH-SY5Y cells 1 to 10 μM PFT-α analog Y-6-159) and, 1 hr. later were exposed to glutamate (SH-SY5Y cells: 100 mM; primary cells: 100 μM, for 24 hr.) or oxidative stress (H2O2: 500 μM, 24 hr.) or no insult for naturally degenerating primary cortical cultures. The percent of neuronal survivals at 24 hr. are shown in Figure 2.

Bottom Line: With parallels to concussive mild traumatic brain injury (mTBI) occurring in humans, anesthetized mice subjected to a single 30 g weight drop mTBI event to the right parietal cortex exhibited significant diffuse neuronal degeneration that was accompanied by delayed impairments in recognition and spatial memory.Cellular studies on human SH-SY5Y cells and rat primary neurons challenged with glutamate excitotoxicity and H2O2 induced oxidative stress, confirmed the ability of PFT-α and a close analog to protect against these TBI associated mechanisms mediating neuronal loss.These studies suggest that p53-dependent apoptotic mechanisms underpin the neuronal and cognitive losses accompanying mTBI, and that these are potentially reversible by p53 inactivation.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy and Anthropology, Sackler School of Medicine and Sagol School of Neuroscience, Tel Aviv University, Tel Aviv, Israel.

ABSTRACT
With parallels to concussive mild traumatic brain injury (mTBI) occurring in humans, anesthetized mice subjected to a single 30 g weight drop mTBI event to the right parietal cortex exhibited significant diffuse neuronal degeneration that was accompanied by delayed impairments in recognition and spatial memory. To elucidate the involvement of reversible p53-dependent apoptosis in this neuronal loss and associated cognitive deficits, mice were subjected to experimental mTBI followed by the systemic administration of the tetrahydrobenzothiazole p53 inactivator, PFT-α, or vehicle. Neuronal loss was quantified immunohistochemically at 72 hr. post-injury by the use of fluoro-Jade B and NeuN within the dentate gyrus on both sides of the brain, and recognition and spatial memory were assessed by novel object recognition and Y-maze paradigms at 7 and 30 days post injury. Systemic administration of a single dose of PFT-α 1 hr. post-injury significantly ameliorated both neuronal cell death and cognitive impairments, which were no different from sham control animals. Cellular studies on human SH-SY5Y cells and rat primary neurons challenged with glutamate excitotoxicity and H2O2 induced oxidative stress, confirmed the ability of PFT-α and a close analog to protect against these TBI associated mechanisms mediating neuronal loss. These studies suggest that p53-dependent apoptotic mechanisms underpin the neuronal and cognitive losses accompanying mTBI, and that these are potentially reversible by p53 inactivation.

Show MeSH
Related in: MedlinePlus