Journal article
Disruption of cell cycle kinetics and cyclin-dependent kinase system by ethanol in cultured cerebellar granule progenitors
Brain research. Developmental brain research, Vol.132(1), pp.47-58
2001
DOI: 10.1016/S0165-3806(01)00294-2
PMID: 11744106
Abstract
An in vitro model of neuronal precursors, primary culture of cerebellar granule progenitors (CGPs), was used to investigate the mechanisms underlying ethanol-induced cell cycle damage. The CGP cultures were generated from 3-day-old rats. Ethanol significantly inhibited the proliferation of the CGPs in culture. Analysis of cell cycle kinetics by a cumulative 5-bromo-2′-deoxyuridine (BrdU) labeling technique demonstrated that ethanol exposure increased the duration of the cell cycle and decreased the growth fraction (the cycling population). The duration of the S-phase and total cell cycle was significantly prolonged by ethanol exposure by 220% and 135%, respectively, while the growth fraction was decreased from 44% in the control groups to 22% in the ethanol-exposed cultures. Cyclin-dependent kinase 2 (Cdk2) is a key protein that regulates both the passage from G1 into S, and the S phase progression. The results from in vitro phosphorylation assay and Western blot demonstrated that ethanol dramatically down-regulated both the activity and the expression of Cdk2. In addition, ethanol significantly decreased the expression of Cyclin A and Cyclin D
2. Further studies using in situ TUNEL assay and DNA fragmentation ELISA showed that ethanol caused a delayed apoptosis, i.e. the ethanol-induced apoptosis was evident only after chronic exposure. On the other hand, ethanol did not affect the necrotic index. In conclusion, ethanol decreases the cycling pool of CGPs by inducing cell cycle delay and promoting apoptosis. Ethanol-mediated disturbance of the cyclin-dependent kinase system may be an important mechanism to account for cell cycle arrest in neuronal precursor cells.
Details
- Title: Subtitle
- Disruption of cell cycle kinetics and cyclin-dependent kinase system by ethanol in cultured cerebellar granule progenitors
- Creators
- Zheng Li - West Virginia UniversityHong Lin - West Virginia UniversityYunfeng Zhu - West Virginia UniversityMei Wang - West Virginia UniversityJia Luo - West Virginia University
- Resource Type
- Journal article
- Publication Details
- Brain research. Developmental brain research, Vol.132(1), pp.47-58
- Publisher
- Elsevier B.V
- DOI
- 10.1016/S0165-3806(01)00294-2
- PMID
- 11744106
- ISSN
- 0165-3806
- Language
- English
- Date published
- 2001
- Academic Unit
- Pathology
- Record Identifier
- 9984186546702771
Metrics
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