Increases in expression of 14-3-3 eta and 14-3-3 zeta transcripts during neuroprotection induced by delta9-tetrahydrocannabinol in AF5 cells.
The molecular mechanisms involved in N-methyl-D-aspartate (NMDA)-induced cell death and Delta9-tetrahydrocannabinol (THC)-induced neuroprotection were investigated in vitro with an AF5 neural progenitor cell line model. By microarray analysis, Ywhah, CK1, Hsp60, Pdcd 4, and Pdcd 7 were identified as being strongly regulated by both NMDA toxicity and THC neuroprotection. The 14-3-3 eta (14-3-3eta; gene symbol Ywhah) and 14-3-3 zeta (14-3-3zeta; gene symbol Ywhaz) transcripts were deceased by NMDA treatment and increased by THC treatment prior to NMDA, as measured by cDNA microarray analysis and quantitative real-time RT-PCR. Other 14-3-3 isoforms were unchanged. Whereas up-regulation of 14-3-3zeta expression was observed 30 min after treatment with THC plus NMDA, down-regulation by NMDA alone was not seen until 16 hr after treatment. By Western blotting, THC increased 14-3-3 protein only in cells that were also treated with NMDA. Overexpression of 14-3-3eta or 14-3-3zeta by transient plasmid transfection increased 14-3-3 protein levels and decreased NMDA-induced cell death. These data suggest that increases in 14-3-3 proteins mediate THC-induced neuroprotection under conditions of NMDA-induced cellular stress.
Chen
Jia
J
Development and Plasticity Section, Cellular Neurobiology Research Branch, Intramural Research Program, National Institute on Drug Abuse, National Institutes of Health, Department of Health and Human Services, Baltimore, MD 21224, USA. jichen@mail.nih.gov
Lee
Chun-Ting
CT
Errico
Stacie L
SL
Becker
Kevin G
KG
Freed
William J
WJ
D048948:14-3-3 Proteins / Q000096:biosynthesis* / Q000235:genetics; D000818:Animals; D002460:Cell Line; D013759:Dronabinol / Q000494:pharmacology*; D008636:Mesencephalon / Q000166:cytology; D016202:N-Methylaspartate / Q000037:antagonists & inhibitors / Q000502:physiology* / Q000633:toxicity; D018696:Neuroprotective Agents / Q000494:pharmacology*; D020411:Oligonucleotide Array Sequence Analysis; D010957:Plasmids; D051381:Rats; D014162:Transfection
D048948:14-3-3 Proteins; D018696:Neuroprotective Agents; D016202:N-Methylaspartate; D013759:Dronabinol