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Functional expression of dihydropyridine-insensitive calcium channels during PC12 cell differentiation by nerve
D M Rausch1, D L Lewis, J L Barker
1Unit on Molecular and Cellular Neurobiology, National Institute of Mental Health, NIH, Bethesda, Maryland 20892.
Abstract:
1. Recombinant retroviruses were used to introduce a temperature-sensitive v-src gene and oncogenic c-Ha-ras into PC12 cells, and stable cell lines expressing these genes were established. 2. As previously reported, expression of v-src (Alema et al., 1985) or c-Ha-ras (Noda et al., 1985) in PC12 cells results in neurite outgrowth resembling that induced by NGF. We report here that v-src but not oncogenic c-Ha-ras induces a stable morphologic neuronal differentiation similar to treatment with NGF. Oncogenic c-Ha-ras-induced neurite outgrowth is not stable with long-term culture, rather the cells revert to an undifferentiated morphology with altered cell cycle kinetics. 3. The stable neuronal phenotype induced by v-src and NGF is characterized by the functional expression of dihydropyridine-insensitive calcium currents.
Insights
The v-src gene induces stable neuronal differentiation in PC12 cells, unlike oncogenic c-Ha-ras, which causes temporary neurite outgrowth. This stable differentiation involves functional calcium currents.
Area of Science:
- Cell biology
- Neuroscience
- Molecular biology
Background:
- PC12 cells are a model system for neuronal differentiation.
- Nerve growth factor (NGF) is a known inducer of neuronal differentiation in PC12 cells.
- Viral oncogenes and mutated cellular oncogenes can influence cell differentiation.
Purpose of the Study:
- To investigate the effects of temperature-sensitive v-src and oncogenic c-Ha-ras on PC12 cell differentiation.
- To compare the stability and characteristics of neuronal differentiation induced by v-src, c-Ha-ras, and NGF.
Main Methods:
- Stable PC12 cell lines expressing temperature-sensitive v-src or oncogenic c-Ha-ras were generated using recombinant retroviruses.
- Morphological changes, including neurite outgrowth, were assessed.
- Cell cycle kinetics were analyzed.
- Functional expression of calcium currents was measured.
Main Results:
- Both v-src and c-Ha-ras induced neurite outgrowth in PC12 cells, mimicking NGF.
- v-src induced stable neuronal differentiation, whereas c-Ha-ras induced transient neurite outgrowth followed by dedifferentiation and altered cell cycle kinetics.
- The stable neuronal phenotype from v-src and NGF was associated with dihydropyridine-insensitive calcium currents.
Conclusions:
- v-src promotes stable neuronal differentiation in PC12 cells, offering a distinct mechanism from NGF.
- Oncogenic c-Ha-ras induces a less stable differentiation response with significant cell cycle alterations.
- The study highlights differential roles of v-src and c-Ha-ras in neuronal differentiation and identifies calcium current expression as a marker for stable neuronal phenotype.