title: Induction of neurite outgrowth in PC12 cells treated with temperature-controlled repeated thermal stimulation.
abstract: To promote the functional restoration of the nervous system following injury, it is necessary to provide optimal extracellular signals that can induce neuronal regenerative activities, particularly neurite formation. This study aimed to examine the regulation of neuritogenesis by temperature-controlled repeated thermal stimulation (TRTS) in rat PC12 pheochromocytoma cells, which can be induced by neurotrophic factors to differentiate into neuron-like cells with elongated neurites. A heating plate was used to apply thermal stimulation, and the correlation of culture medium temperature with varying surface temperature of the heating plate was monitored. Plated PC12 cells were exposed to TRTS at two different temperatures via heating plate (preset surface temperature of the heating plate, 39.5°C or 42°C) in growth or differentiating medium for up to 18 h per day. We then measured the extent of growth, neuritogenesis, or acetylcholine esterase (AChE) activity (a neuronal marker). To analyze the mechanisms underlying the effects of TRTS on these cells, we examined changes in intracellular signaling using the following: tropomyosin-related kinase A inhibitor GW441756; p38 mitogen-activated protein kinase (MAPK) inhibitor SB203580; and MAPK/extracellular signal-regulated kinase (ERK) kinase (MEK) inhibitor U0126 with its inactive analog, U0124, as a control. While a TRTS of 39.5°C did not decrease the growth rate of cells in the cell growth assay, it did increase the number of neurite-bearing PC12 cells and AChE activity without the addition of other neuritogenesis inducers. Furthermore, U0126, and SB203580, but not U0124 and GW441756, considerably inhibited TRTS-induced neuritogenesis. These results suggest that TRTS can induce neuritogenesis and that participation of both the ERK1/2 and p38 MAPK signaling pathways is required for TRTS-dependent neuritogenesis in PC12 cells. Thus, TRTS may be an effective technique for regenerative neuromedicine.
authors: Division of Oral Physiology, Tohoku University Graduate School of Dentistry, Sendai city, Miyagi, Japan.
authors: Tada-aki
authors: TA
authors: Kudo
authors: Liaison Center for Innovative Dentistry, Tohoku University Graduate School of Dentistry, Sendai city, Miyagi, Japan; Tohoku University Graduate School of Biomedical Engineering, Sendai city, Miyagi, Japan.
authors: Hiroyasu
authors: H
authors: Kanetaka
authors: Cell Resource Center for Biomedical Research, Institute of Development, Aging and Cancer, Tohoku University, Sendai city, Miyagi, Japan.
authors: Kentaro
authors: K
authors: Mochizuki
authors: Tohoku University Graduate School of Biomedical Engineering, Sendai city, Miyagi, Japan.
authors: Kanako
authors: K
authors: Tominami
authors: Division of Oral Dysfunction Science, Tohoku University Graduate School of Dentistry, Sendai city, Miyagi, Japan.
authors: Shoko
authors: S
authors: Nunome
authors: Department of Physical Medicine and Rehabilitation, Tohoku University Graduate School of Medicine, Sendai city, Miyagi, Japan.
authors: Genji
authors: G
authors: Abe
authors: Institute of Fluid Science, Tohoku University, Sendai city, Miyagi, Japan.
authors: Hiroyuki
authors: H
authors: Kosukegawa
authors: IFG Co., Ltd, Sendai city, Miyagi, Japan.
authors: Toshihiko
authors: T
authors: Abe
authors: IFG Co., Ltd, Sendai city, Miyagi, Japan.
authors: Hitoshi
authors: H
authors: Mori
authors: IFG Co., Ltd, Sendai city, Miyagi, Japan.
authors: Kazumi
authors: K
authors: Mori
authors: Institute of Fluid Science, Tohoku University, Sendai city, Miyagi, Japan.
authors: Toshiyuki
authors: T
authors: Takagi
authors: Tohoku University Graduate School of Biomedical Engineering, Sendai city, Miyagi, Japan; Department of Physical Medicine and Rehabilitation, Tohoku University Graduate School of Medicine, Sendai city, Miyagi, Japan.
authors: Shin-ichi
authors: S
authors: Izumi
mesh_terms: D000110:Acetylcholinesterase / Q000378:metabolism; D000818:Animals; D002070:Butadienes / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects; D019439:Chromaffin Cells / Q000166:cytology / Q000378:metabolism; D007093:Imidazoles / Q000494:pharmacology; D007211:Indoles / Q000494:pharmacology*; D016501:Neurites / Q000378:metabolism*; D055495:Neurogenesis / Q000187:drug effects; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D009570:Nitriles / Q000494:pharmacology; D016716:PC12 Cells; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D011725:Pyridines / Q000494:pharmacology; D051381:Rats; D020917:Receptor, trkA / Q000037:antagonists & inhibitors; D015398:Signal Transduction / Q000187:drug effects; D013696:Temperature
chemical_list: D002070:Butadienes; C000606649:GW 441756; D007093:Imidazoles; D007211:Indoles; D009570:Nitriles; D047428:Protein Kinase Inhibitors; D011725:Pyridines; C113580:U 0126; D020917:Receptor, trkA; D000110:Acetylcholinesterase; C093642:SB 203580