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Tau and caspase 3 as targets for neuroprotection.

Idan-Feldman A, Ostritsky R, Gozes I - Int J Alzheimers Dis (2012)

Bottom Line: The peptide drug candidate NAP (davunetide) has demonstrated protective effects in various in vivo and in vitro models of neurodegeneration.NAP was shown to reduce tau hyperphosphorylation as well as to prevent caspase-3 activation and cytochrome-3 release from mitochondria, both characteristic of apoptotic cell death.The purpose of this study was to evaluate the effect of NAP on tau hyperphosphorylation and caspase activity in the same biological system.

View Article: PubMed Central - PubMed

Affiliation: The Adams Super Center for Brain Studies, The Lily and Avraham Gildor Chair for The Investigation of Growth Factors, The Elton Laboratory for Molecular Neuroendocrinology, and Department of Human Molecular Genetics and Biochemistry, Sagol School of Neuroscience, Sackler Faculty of Medicine, Tel Aviv University, 69978 Tel Aviv, Israel.

ABSTRACT
The peptide drug candidate NAP (davunetide) has demonstrated protective effects in various in vivo and in vitro models of neurodegeneration. NAP was shown to reduce tau hyperphosphorylation as well as to prevent caspase-3 activation and cytochrome-3 release from mitochondria, both characteristic of apoptotic cell death. Recent studies suggest that caspases may play a role in tau pathology. The purpose of this study was to evaluate the effect of NAP on tau hyperphosphorylation and caspase activity in the same biological system. Our experimental setup used primary neuronal cultures subjected to oxygen-glucose deprivation (OGD), with and without NAP or caspase inhibitor. Cell viability was assessed by measuring mitochondrial activity (MTS assay), and immunoblots were used for analyzing protein level. It was shown that apoptosis was responsible for all cell death occurring following ischemia, and NAP treatment showed a concentration-dependent protection from cell death. Ischemia caused an increase in the levels of active caspase-3 and hyperphosphorylated tau, both of which were prevented by either NAP or caspase-inhibitor treatment. Our data suggest that, in this model system, caspase activation may be an upstream event to tau hyperphosphorylation, although additional studies will be required to fully elucidate the cascade of events.

No MeSH data available.


Related in: MedlinePlus

Culture purity: at 5-6 DIV, cells were fixed and immunofluorescence staining was performed using the astrocyte marker GFAP (green), the neuronal marker NeuN (red), and DAPI stain (blue) for nuclei. Quantification of neuronal cells was done using 10 random fields from each of 3 experiments (×20 magnification). 95.9 ± 0.96% of the total cells (DAPI stain) were recognized as neurons (NeuN positive, GFAP negative).
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fig1: Culture purity: at 5-6 DIV, cells were fixed and immunofluorescence staining was performed using the astrocyte marker GFAP (green), the neuronal marker NeuN (red), and DAPI stain (blue) for nuclei. Quantification of neuronal cells was done using 10 random fields from each of 3 experiments (×20 magnification). 95.9 ± 0.96% of the total cells (DAPI stain) were recognized as neurons (NeuN positive, GFAP negative).

Mentions: Culture purity was analyzed using immunofluorescent staining with cell-specific markers. After 5-6 DIV (days in vitro), 95.9 ± 0.96% (SEM) of the cells (counted using nuclear DAPI stain) were positive for the neuronal NeuN marker and negative for the astrocytic marker, GFAP. A representative immunefluorescence stain in Figure 1 shows neuronal cells stained with NeuN in red, and astrocytes stained with GFAP in green.


Tau and caspase 3 as targets for neuroprotection.

Idan-Feldman A, Ostritsky R, Gozes I - Int J Alzheimers Dis (2012)

Culture purity: at 5-6 DIV, cells were fixed and immunofluorescence staining was performed using the astrocyte marker GFAP (green), the neuronal marker NeuN (red), and DAPI stain (blue) for nuclei. Quantification of neuronal cells was done using 10 random fields from each of 3 experiments (×20 magnification). 95.9 ± 0.96% of the total cells (DAPI stain) were recognized as neurons (NeuN positive, GFAP negative).
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3369463&req=5

fig1: Culture purity: at 5-6 DIV, cells were fixed and immunofluorescence staining was performed using the astrocyte marker GFAP (green), the neuronal marker NeuN (red), and DAPI stain (blue) for nuclei. Quantification of neuronal cells was done using 10 random fields from each of 3 experiments (×20 magnification). 95.9 ± 0.96% of the total cells (DAPI stain) were recognized as neurons (NeuN positive, GFAP negative).
Mentions: Culture purity was analyzed using immunofluorescent staining with cell-specific markers. After 5-6 DIV (days in vitro), 95.9 ± 0.96% (SEM) of the cells (counted using nuclear DAPI stain) were positive for the neuronal NeuN marker and negative for the astrocytic marker, GFAP. A representative immunefluorescence stain in Figure 1 shows neuronal cells stained with NeuN in red, and astrocytes stained with GFAP in green.

Bottom Line: The peptide drug candidate NAP (davunetide) has demonstrated protective effects in various in vivo and in vitro models of neurodegeneration.NAP was shown to reduce tau hyperphosphorylation as well as to prevent caspase-3 activation and cytochrome-3 release from mitochondria, both characteristic of apoptotic cell death.The purpose of this study was to evaluate the effect of NAP on tau hyperphosphorylation and caspase activity in the same biological system.

View Article: PubMed Central - PubMed

Affiliation: The Adams Super Center for Brain Studies, The Lily and Avraham Gildor Chair for The Investigation of Growth Factors, The Elton Laboratory for Molecular Neuroendocrinology, and Department of Human Molecular Genetics and Biochemistry, Sagol School of Neuroscience, Sackler Faculty of Medicine, Tel Aviv University, 69978 Tel Aviv, Israel.

ABSTRACT
The peptide drug candidate NAP (davunetide) has demonstrated protective effects in various in vivo and in vitro models of neurodegeneration. NAP was shown to reduce tau hyperphosphorylation as well as to prevent caspase-3 activation and cytochrome-3 release from mitochondria, both characteristic of apoptotic cell death. Recent studies suggest that caspases may play a role in tau pathology. The purpose of this study was to evaluate the effect of NAP on tau hyperphosphorylation and caspase activity in the same biological system. Our experimental setup used primary neuronal cultures subjected to oxygen-glucose deprivation (OGD), with and without NAP or caspase inhibitor. Cell viability was assessed by measuring mitochondrial activity (MTS assay), and immunoblots were used for analyzing protein level. It was shown that apoptosis was responsible for all cell death occurring following ischemia, and NAP treatment showed a concentration-dependent protection from cell death. Ischemia caused an increase in the levels of active caspase-3 and hyperphosphorylated tau, both of which were prevented by either NAP or caspase-inhibitor treatment. Our data suggest that, in this model system, caspase activation may be an upstream event to tau hyperphosphorylation, although additional studies will be required to fully elucidate the cascade of events.

No MeSH data available.


Related in: MedlinePlus