Protein kinase C δ mediates cerebral reperfusion injury In Vivo

Rachel Bright, Ami Raval, Jeffrey M. Dembner, Miguel Perez-Pinzon, Gary K. Steinberg, Midori A. Yenari, Daria Mochly-Rosen

Research output: Contribution to journalArticle

156 Citations (Scopus)

Abstract

Protein kinase C (PKC) has been implicated in mediating ischemic and reperfusion damage in multiple organs. However, conflicting reports exist on the role of individual PKC isozymes in cerebral ischemic injury. Using a peptide inhibitor selective for δPKC, δV1-1, we found that δPKC inhibition reduced cellular injury in a rat hippocampal slice model of cerebral ischemia [oxygen-glucose deprivation (OGD)] when present both during OGD and for the first 3 hr of reperfusion. We next demonstrated peptide delivery to the brain parenchyma after in vivo delivery by detecting biotin-conjugated δV1-1 and by measuring inhibition of intracellular δPKC translocation, an indicator of δPKC activity. Delivery of δV1-1 decreased infarct size in an in vivo rat stroke model of transient middle cerebral artery occlusion. Importantly, δV1-1 had no effect when delivered immediately before ischemia. However, delivery at the onset, at 1 hr, or at 6 hr of reperfusion reduced injury by 68, 47, and 58%, respectively. Previous work has implicated δPKC in mediating apoptotic processes. We therefore determined whether δPKC inhibition altered apoptotic cell death or cell survival pathways in our models. We found that δV1-1 reduced numbers of terminal deoxynucleotidyl transferase-mediated biotinylated UTP nick end labeling-positive cells, indicating decreased apoptosis, increased levels of phospho-Akt, a kinase involved in cell survival pathways, and inhibited BAD (Bcl-2-associated death protein) protein translocation from the cell cytosol to the membrane, indicating inhibition of proapoptotic signaling. These data support a deleterious role for δPKC during reperfusion and suggest that δV1-1 delivery, even hours after commencement of reperfusion, may provide a therapeutic advantage after cerebral ischemia.

Original languageEnglish
Pages (from-to)6880-6888
Number of pages9
JournalJournal of Neuroscience
Volume24
Issue number31
DOIs
StatePublished - Aug 4 2004

Fingerprint

Reperfusion Injury
Protein Kinase C
Reperfusion
Brain Ischemia
Cell Survival
bcl-Associated Death Protein
Oxygen
Glucose
Peptides
Uridine Triphosphate
DNA Nucleotidylexotransferase
Middle Cerebral Artery Infarction
Wounds and Injuries
Protein Transport
Biotin
Cytosol
Isoenzymes
Cell Death
Phosphotransferases
Ischemia

Keywords

  • Hippocampus
  • Ischemia
  • Neuroprotection
  • OGD
  • Peptide inhibitor
  • PKC

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Bright, R., Raval, A., Dembner, J. M., Perez-Pinzon, M., Steinberg, G. K., Yenari, M. A., & Mochly-Rosen, D. (2004). Protein kinase C δ mediates cerebral reperfusion injury In Vivo. Journal of Neuroscience, 24(31), 6880-6888. https://doi.org/10.1523/JNEUROSCI.4474-03.2004

Protein kinase C δ mediates cerebral reperfusion injury In Vivo. / Bright, Rachel; Raval, Ami; Dembner, Jeffrey M.; Perez-Pinzon, Miguel; Steinberg, Gary K.; Yenari, Midori A.; Mochly-Rosen, Daria.

In: Journal of Neuroscience, Vol. 24, No. 31, 04.08.2004, p. 6880-6888.

Research output: Contribution to journalArticle

Bright, R, Raval, A, Dembner, JM, Perez-Pinzon, M, Steinberg, GK, Yenari, MA & Mochly-Rosen, D 2004, 'Protein kinase C δ mediates cerebral reperfusion injury In Vivo', Journal of Neuroscience, vol. 24, no. 31, pp. 6880-6888. https://doi.org/10.1523/JNEUROSCI.4474-03.2004
Bright, Rachel ; Raval, Ami ; Dembner, Jeffrey M. ; Perez-Pinzon, Miguel ; Steinberg, Gary K. ; Yenari, Midori A. ; Mochly-Rosen, Daria. / Protein kinase C δ mediates cerebral reperfusion injury In Vivo. In: Journal of Neuroscience. 2004 ; Vol. 24, No. 31. pp. 6880-6888.
@article{92a2ffff80944a7884852e14e5b79a6b,
title = "Protein kinase C δ mediates cerebral reperfusion injury In Vivo",
abstract = "Protein kinase C (PKC) has been implicated in mediating ischemic and reperfusion damage in multiple organs. However, conflicting reports exist on the role of individual PKC isozymes in cerebral ischemic injury. Using a peptide inhibitor selective for δPKC, δV1-1, we found that δPKC inhibition reduced cellular injury in a rat hippocampal slice model of cerebral ischemia [oxygen-glucose deprivation (OGD)] when present both during OGD and for the first 3 hr of reperfusion. We next demonstrated peptide delivery to the brain parenchyma after in vivo delivery by detecting biotin-conjugated δV1-1 and by measuring inhibition of intracellular δPKC translocation, an indicator of δPKC activity. Delivery of δV1-1 decreased infarct size in an in vivo rat stroke model of transient middle cerebral artery occlusion. Importantly, δV1-1 had no effect when delivered immediately before ischemia. However, delivery at the onset, at 1 hr, or at 6 hr of reperfusion reduced injury by 68, 47, and 58{\%}, respectively. Previous work has implicated δPKC in mediating apoptotic processes. We therefore determined whether δPKC inhibition altered apoptotic cell death or cell survival pathways in our models. We found that δV1-1 reduced numbers of terminal deoxynucleotidyl transferase-mediated biotinylated UTP nick end labeling-positive cells, indicating decreased apoptosis, increased levels of phospho-Akt, a kinase involved in cell survival pathways, and inhibited BAD (Bcl-2-associated death protein) protein translocation from the cell cytosol to the membrane, indicating inhibition of proapoptotic signaling. These data support a deleterious role for δPKC during reperfusion and suggest that δV1-1 delivery, even hours after commencement of reperfusion, may provide a therapeutic advantage after cerebral ischemia.",
keywords = "Hippocampus, Ischemia, Neuroprotection, OGD, Peptide inhibitor, PKC",
author = "Rachel Bright and Ami Raval and Dembner, {Jeffrey M.} and Miguel Perez-Pinzon and Steinberg, {Gary K.} and Yenari, {Midori A.} and Daria Mochly-Rosen",
year = "2004",
month = "8",
day = "4",
doi = "10.1523/JNEUROSCI.4474-03.2004",
language = "English",
volume = "24",
pages = "6880--6888",
journal = "Journal of Neuroscience",
issn = "0270-6474",
publisher = "Society for Neuroscience",
number = "31",

}

TY - JOUR

T1 - Protein kinase C δ mediates cerebral reperfusion injury In Vivo

AU - Bright, Rachel

AU - Raval, Ami

AU - Dembner, Jeffrey M.

AU - Perez-Pinzon, Miguel

AU - Steinberg, Gary K.

AU - Yenari, Midori A.

AU - Mochly-Rosen, Daria

PY - 2004/8/4

Y1 - 2004/8/4

N2 - Protein kinase C (PKC) has been implicated in mediating ischemic and reperfusion damage in multiple organs. However, conflicting reports exist on the role of individual PKC isozymes in cerebral ischemic injury. Using a peptide inhibitor selective for δPKC, δV1-1, we found that δPKC inhibition reduced cellular injury in a rat hippocampal slice model of cerebral ischemia [oxygen-glucose deprivation (OGD)] when present both during OGD and for the first 3 hr of reperfusion. We next demonstrated peptide delivery to the brain parenchyma after in vivo delivery by detecting biotin-conjugated δV1-1 and by measuring inhibition of intracellular δPKC translocation, an indicator of δPKC activity. Delivery of δV1-1 decreased infarct size in an in vivo rat stroke model of transient middle cerebral artery occlusion. Importantly, δV1-1 had no effect when delivered immediately before ischemia. However, delivery at the onset, at 1 hr, or at 6 hr of reperfusion reduced injury by 68, 47, and 58%, respectively. Previous work has implicated δPKC in mediating apoptotic processes. We therefore determined whether δPKC inhibition altered apoptotic cell death or cell survival pathways in our models. We found that δV1-1 reduced numbers of terminal deoxynucleotidyl transferase-mediated biotinylated UTP nick end labeling-positive cells, indicating decreased apoptosis, increased levels of phospho-Akt, a kinase involved in cell survival pathways, and inhibited BAD (Bcl-2-associated death protein) protein translocation from the cell cytosol to the membrane, indicating inhibition of proapoptotic signaling. These data support a deleterious role for δPKC during reperfusion and suggest that δV1-1 delivery, even hours after commencement of reperfusion, may provide a therapeutic advantage after cerebral ischemia.

AB - Protein kinase C (PKC) has been implicated in mediating ischemic and reperfusion damage in multiple organs. However, conflicting reports exist on the role of individual PKC isozymes in cerebral ischemic injury. Using a peptide inhibitor selective for δPKC, δV1-1, we found that δPKC inhibition reduced cellular injury in a rat hippocampal slice model of cerebral ischemia [oxygen-glucose deprivation (OGD)] when present both during OGD and for the first 3 hr of reperfusion. We next demonstrated peptide delivery to the brain parenchyma after in vivo delivery by detecting biotin-conjugated δV1-1 and by measuring inhibition of intracellular δPKC translocation, an indicator of δPKC activity. Delivery of δV1-1 decreased infarct size in an in vivo rat stroke model of transient middle cerebral artery occlusion. Importantly, δV1-1 had no effect when delivered immediately before ischemia. However, delivery at the onset, at 1 hr, or at 6 hr of reperfusion reduced injury by 68, 47, and 58%, respectively. Previous work has implicated δPKC in mediating apoptotic processes. We therefore determined whether δPKC inhibition altered apoptotic cell death or cell survival pathways in our models. We found that δV1-1 reduced numbers of terminal deoxynucleotidyl transferase-mediated biotinylated UTP nick end labeling-positive cells, indicating decreased apoptosis, increased levels of phospho-Akt, a kinase involved in cell survival pathways, and inhibited BAD (Bcl-2-associated death protein) protein translocation from the cell cytosol to the membrane, indicating inhibition of proapoptotic signaling. These data support a deleterious role for δPKC during reperfusion and suggest that δV1-1 delivery, even hours after commencement of reperfusion, may provide a therapeutic advantage after cerebral ischemia.

KW - Hippocampus

KW - Ischemia

KW - Neuroprotection

KW - OGD

KW - Peptide inhibitor

KW - PKC

UR - http://www.scopus.com/inward/record.url?scp=3843146318&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=3843146318&partnerID=8YFLogxK

U2 - 10.1523/JNEUROSCI.4474-03.2004

DO - 10.1523/JNEUROSCI.4474-03.2004

M3 - Article

C2 - 15295022

AN - SCOPUS:3843146318

VL - 24

SP - 6880

EP - 6888

JO - Journal of Neuroscience

JF - Journal of Neuroscience

SN - 0270-6474

IS - 31

ER -