Abstract
Excitotoxicity via N-methyl-D-aspartate receptors (NMDA-r) is a key mechanism of neurodegeneration following ischemia and other brain pathologies. We here describe the use of a fast, reliable and easily reproducible in vitro model of excitotoxicity induced with NMDA stimulation of cortical neurons. Such a protocol can be used to study the intracellular pathways involved in NMDA-induced death. Here wefocused on JNK signaling, a pathway that plays a determinant role in excitotoxic death of cortical neurons. Our proposed model can additionally be used to test different drugs designed to prevent excitotoxic neuronal death. As an example we describe the use of the cell permeable specific JNK inhibitor peptide as a neuroprotective agent. This chapter will give methodological suggestions to help researchers prepare cortical neurons, to develop an in vitro model of neuronal death by stimulating neurons with NMDA, to quantify neuronal death in vitro using LDH assay and propidium staining, to analyze the intracellular pathways involved (JNK pathway) in neuronal death, and to perform protocols of neuroprotection.
| Original language | English |
|---|---|
| Title of host publication | Neuromethods |
| Publisher | Humana Press Inc. |
| Pages | 189-202 |
| Number of pages | 14 |
| Volume | 68 |
| DOIs | |
| Publication status | Published - 2012 |
Publication series
| Name | Neuromethods |
|---|---|
| Volume | 68 |
| ISSN (Print) | 08932336 |
| ISSN (Electronic) | 19406045 |
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Keywords
- Cell permeable JNK inhibitor peptide
- D-JNKI1
- JNK
- Neuronal death
- Neuroprotection
- NMDA-toxicity
- Signaling pathway
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Pharmacology, Toxicology and Pharmaceutics(all)
- Neuroscience(all)
- Psychiatry and Mental health
Cite this
Cortical neurons culture to study C-Jun N-Terminal kinase signaling pathway. / Sclip, Alessandra; Antoniou, Xanthi; Borsello, Tiziana.
Neuromethods. Vol. 68 Humana Press Inc., 2012. p. 189-202 (Neuromethods; Vol. 68).Research output: Chapter in Book/Report/Conference proceeding › Chapter
}
TY - CHAP
T1 - Cortical neurons culture to study C-Jun N-Terminal kinase signaling pathway
AU - Sclip, Alessandra
AU - Antoniou, Xanthi
AU - Borsello, Tiziana
PY - 2012
Y1 - 2012
N2 - Excitotoxicity via N-methyl-D-aspartate receptors (NMDA-r) is a key mechanism of neurodegeneration following ischemia and other brain pathologies. We here describe the use of a fast, reliable and easily reproducible in vitro model of excitotoxicity induced with NMDA stimulation of cortical neurons. Such a protocol can be used to study the intracellular pathways involved in NMDA-induced death. Here wefocused on JNK signaling, a pathway that plays a determinant role in excitotoxic death of cortical neurons. Our proposed model can additionally be used to test different drugs designed to prevent excitotoxic neuronal death. As an example we describe the use of the cell permeable specific JNK inhibitor peptide as a neuroprotective agent. This chapter will give methodological suggestions to help researchers prepare cortical neurons, to develop an in vitro model of neuronal death by stimulating neurons with NMDA, to quantify neuronal death in vitro using LDH assay and propidium staining, to analyze the intracellular pathways involved (JNK pathway) in neuronal death, and to perform protocols of neuroprotection.
AB - Excitotoxicity via N-methyl-D-aspartate receptors (NMDA-r) is a key mechanism of neurodegeneration following ischemia and other brain pathologies. We here describe the use of a fast, reliable and easily reproducible in vitro model of excitotoxicity induced with NMDA stimulation of cortical neurons. Such a protocol can be used to study the intracellular pathways involved in NMDA-induced death. Here wefocused on JNK signaling, a pathway that plays a determinant role in excitotoxic death of cortical neurons. Our proposed model can additionally be used to test different drugs designed to prevent excitotoxic neuronal death. As an example we describe the use of the cell permeable specific JNK inhibitor peptide as a neuroprotective agent. This chapter will give methodological suggestions to help researchers prepare cortical neurons, to develop an in vitro model of neuronal death by stimulating neurons with NMDA, to quantify neuronal death in vitro using LDH assay and propidium staining, to analyze the intracellular pathways involved (JNK pathway) in neuronal death, and to perform protocols of neuroprotection.
KW - Cell permeable JNK inhibitor peptide
KW - D-JNKI1
KW - JNK
KW - Neuronal death
KW - Neuroprotection
KW - NMDA-toxicity
KW - Signaling pathway
UR - http://www.scopus.com/inward/record.url?scp=84979284931&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84979284931&partnerID=8YFLogxK
U2 - 10.1007/978-1-61779-824-5_10
DO - 10.1007/978-1-61779-824-5_10
M3 - Chapter
AN - SCOPUS:84979284931
VL - 68
T3 - Neuromethods
SP - 189
EP - 202
BT - Neuromethods
PB - Humana Press Inc.
ER -