The Effects of LPS With or Without Forskolin on RT4-D6P2T Cell Morphology
Schwann cells can be used as a model to study different signaling pathways involved during injury and inflammation in the peripheral nervous system. For this study, the immortalized rat RT4-D6P2T Schwannoma cell line from ATCC (#CRL-2768) was used. The cells were treated with different doses of lipopolysaccharide (LPS) with or without 2 uM of forskolin for 3 hours to determine the effects of LPS with or without forskolin on Schwann cell morphology. LPS is a cell wall immunostimulatory component found in Gram-negative bacteria. It activates the nuclear factor kappa B (NF-KB) pathway, which plays a major role in inflammation through the production of inflammatory cytokines, such as tumor necrosis factor alpha (TNF-alpha). Forskolin is a root extract from the Indian plant Coleus forskohlii that activates the cAMP pathway, which plays a major role in cell proliferation, axonal regeneration, inflammation, and more. Although it is well-known that both the NF-KB and cAMP pathways are involved in inflammation, not much is known regarding the effects of NF-KB activation, with or without cAMP activation, on Schwann cell morphology. It was hypothesized that cells treated with LPS and forskolin would have significant morphological differences compared to cells treated with LPS only. Before and after a 3-hour incubation with the different treatment combinations, images of the cell cultures were taken at a magnification of 10X using a Zeiss Primovert iLED microscope equipped with Zeiss Axiocam 208 color microscope camera. The provided figure shows representative images of the cells from three independent experiments. Unexpectedly, the cells did not appear to have any obvious morphological changes after a 3-hour incubation with each of the treatments. However, there were large gaps on the surface of the culture flasks where the cells used to be. There is not enough information to determine what exactly caused these gaps. Some possibilities include apoptosis/cell death or cell retraction/migration. Further studies need to be done before conclusions can be made.