Bcl-2 overexpression disrupts the morphology of PC12 cells through reduced ERK activation
Department of Pediatrics, University of Pittsburgh, Pittsburgh, Pennsylvania, United States Brain Research
(Impact Factor: 2.84).
10/2006; 1112(1):46-55. DOI: 10.1016/j.brainres.2006.07.017
Bcl-2 has been hypothesized to regulate many cellular functions in addition to its well-characterized role in the prevention of programmed cell death. To understand the role of Bcl-2 in regulating cell morphology and to explore the mechanism of this effect, we examined the effects of Bcl-2 overexpression on the morphology of PC12 cells in culture. We demonstrate that the overexpression of Bcl-2 in PC12 cells results in altered cell morphology and reduced actin expression. Analysis of extracellular signal-regulated kinase (ERK) 1/2 phosphorylation reveals that the morphological changes seen after bcl-2 transfection are associated with reduced ERK activation. Treatment of control (mock-transfected) PC12 cells with the mitogen-activated ERK-activating kinase (MEK) inhibitor PD98059 converts their flat, process-bearing morphology into the rounded, process-free morphology of bcl-2-transfected cells, further confirming the association of ERK activation with altered cell shape. In conclusion, the present study describes a novel function of Bcl-2 in regulating cell shape through reduced ERK activation.
Available from: tu-dortmund.de
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ABSTRACT: The mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) pathway is important for both long-term survival and timing of the progression of oligodendrocyte differentiation. Oligodendroglial cells treated with MEK inhibitor were distinguished by using stage specific markers: NG2 proteoglycan, A2B5, 2'3'nucleotide-cyclic 3'phosphodiesterase (CNPase) and myelin basic protein (MBP), and classified according to their morphology into different developmental stages. Treatment significantly increased the number of cells with more immature morphologies and decreased the number of mature cells. Furthermore, it increased the number of rounded cells that could not be classified into any of the oligodendroglial developmental stages. The strongest effects were usually observed shortly after treatment. Rounded cells were CNPase/MBP positive and they were not stained by anti-NG2 or A2B5, indicating that they were mature cells unable either to extend and/or to maintain their processes. These data showed an effect of the MAPK/ERK pathway on oligodendroglial branching, with possible consequences for the formation of the myelin sheath.
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ABSTRACT: A variety of cell lines are often used to simulate the studies associated with the cell products, genetics, cell–cell interaction, environmental interaction, intracellular flux and intracellular activity. Among these cell lines, the pheochromocytoma (PC12 for short) cell line is the one that is widely applied in in vitro study for investigating the neuronal apoptosis, neurochemistry, and neuronal differentiation. The development of an appropriate cell cultivation platform as well as the real-time image acquisition and analysis system becomes an important issue. The purpose of this paper is to develop a long-term and low-cost incubation system, including microscopic image analysis system for morphological analysis. The proposed incubation system can acquire cell image for analysis during cell cultivation. From the experimental results, the temperature of incubation system is found to be in the range of 37.3 ± 1.3°C, and the lower levels of the relative humidity vary within 89.7 ± 1.8%. The CO2 concentration is controlled at 5.29 ± 0.71%. The cell number is increased from 1.00 × 105 to 4.25 × 105 cells/ml after 48 h of culture. The cell viability test shows that this developed incubation system works well for PC12 cells cultivation; it can also acquire and segment the cell image during the cell cultivation. Besides, in the image analysis software design, the region-growing and modified blob labeling techniques are also employed to segment the PC12 cell image under varied culture conditions for later morphological and feature quantification analysis.
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