Induction of Neurite Outgrowth in PC12 Cells

Like all organs in the body, the brain is subject to injury. However, unlike in other organs, brain injury is not something that a little rest, some medication or a minor surgery can help to remedy. Being far more complex in structure and organization, efforts to repair the brain are subsequently more complicated themselves. One of the more novel methods for repairing neurons is to use rat PC12 pheochromocytoma cells, which have been found to differentiate into neuron-like cells with elongated neurites. Treatment of PC12 cells with various soluble factors, such as Nerve Growth Factor (NGF) and bone morphogenetic proteins (BMPs), has been shown to stimulate PC12 cells into becoming neuron-like cells. Specifically, when PC12 cells differentiate following exposure to NGF or NGF-like compounds, they stop proliferating, show increased acetylcholinesterase activity, and become electrically excitable. Kudo et al. from the Tohoku University Graduate School of Dentistry in Japan set out to study this and the effect of temperature-controlled repeated thermal stimulation (TRTS) in generating neural regrowth.

One of the reasons for looking at the role of TRTS in neurogenesis comes from the success of the method as an alternative and safe cancer therapy. However, no researcher had looked at the role of TRTS in neuron differentiation and proliferation. To understand this, Kudo and his team needed to measure the AChE activity -a biochemical marker for neural differentiation. To do this, they used the Amplite^® Colorimetric Acetylcholinesterase Assay Kit. One of the reasons for using this kit is due to the pinpoint accuracy the tool provides. It uses DTNB to measure thiocholine produced from the hydrolysis of acetylthiocholine by AChE in blood, in cell extracts and other solutions. Additionally, its chlorometric one-step assay makes it possible to detect as little as .1 mU AChE in a 100 µL assay volume. This helps researchers get clear indicators of even the smallest amounts of AChE, which allows for more accurate readings and more robust results.

At the conclusion of their study, Kudo and his colleagues found a non-cytotoxic TRTS treatment can induce neuritogenesis without supplementation of other neurotrophic factors, and requires participation of MEK-ERK1/2 and p38 MAPK signaling, similar to BMP4. This is significant as it indicates the prospective role TRTS can play in helping to stimulate neural proliferation and differentiation. Crucial to this study was the ability to measure AChE, as a marker of neural differentiation. Without tools such as the Amplite^® Colorimetric Acetylcholinesterase Assay Kit, studies such as these would not be possible, nor would they be as reliable. Being able to closely examine AChE activity allows for researchers to have greater confidence in their results, producing less controversy and allowing for the medical community to keep moving towards the development of novel and effective treatments for the most damaging brain injuries.

References

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1. Kudo, Tada-aki, et al. "Induction of Neurite Outgrowth in PC12 Cells Treated with Temperature-Controlled Repeated Thermal Stimulation." PloS one 10.4 (2015): e0124024.