AI Article Synopsis
- The study aimed to investigate how 5-lipoxygenase (5-LOX) moves within PC12 cells after injuries, using a fluorescent tagging method with green fluorescence protein (GFP)/5-LOX.
- PC12 cells were genetically modified to express either GFP/5-LOX or GFP alone, and their reactions to various treatments (like oxygen-glucose deprivation and H2O2) were examined under a fluorescence microscope.
- Results showed that GFP/5-LOX mostly stayed in the nucleus and moved to the nuclear membrane after certain treatments, whereas the control cells and other treatment conditions did not show this translocation, indicating different responses of 5-LOX.
Article Abstract
Objective: To evaluate the translocation of 5-lipoxygenase (5-LOX)) after injuries by transfection with green fluorescence protein (GFP)/5-LOX in PC12 cells.
Methods: PC12 cells were stably transfected with pEGFP-C2/5-LOX (GFP/5-LOX) or pEGFP-C2 vectors (control). After treatment with oxygen-glucose deprivation (OGD), H(2)O(2) or NMDA, GFP/5-LOX localization in the cells was observed under a fluorescence microscope. Wild-type 5-LOX was determined by immunostaining after the treatment.
Result: In the GFP/5-LOX-transfected cells, GFP/5-LOX was primarily localized in the nucleus; while in the GFP-transfected cells, GFP was localized in both the cytoplasm and nucleus. After OGD and H(2)O(2) treatments, GFP/5-LOX was translocated to the nuclear membrane in 50.6 % and 57.7% cells respectively. However, after NMDA treatment or in GFP-transfected cells, no translocation was observed. Wild-type 5-LOX was distributed in the nuclei and cytoplasm, and all the 3 treatments induced 5-LOX translocation to the nuclear membrane.
Conclusion: In the PC12 cells stably transfected with GFP/5-LOX, GFP/5-LOX is primarily distributed in the nuclei; the OGD-, H(2)O(2)- and NMDA-induced 5-LOX translocation exhibits different properties.
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| http://dx.doi.org/10.3785/j.issn.1008-9292.2007.02.002 | DOI Listing |
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