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Induction of Neurite Outgrowth in PC12 Cells Treated with Temperature-Controlled Repeated Thermal Stimulation

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-control...

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Autores principales: Kudo, Tada-aki, Kanetaka, Hiroyasu, Mochizuki, Kentaro, Tominami, Kanako, Nunome, Shoko, Abe, Genji, Kosukegawa, Hiroyuki, Abe, Toshihiko, Mori, Hitoshi, Mori, Kazumi, Takagi, Toshiyuki, Izumi, Shin-ichi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4399938/
https://www.ncbi.nlm.nih.gov/pubmed/25879210
http://dx.doi.org/10.1371/journal.pone.0124024
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author Kudo, Tada-aki
Kanetaka, Hiroyasu
Mochizuki, Kentaro
Tominami, Kanako
Nunome, Shoko
Abe, Genji
Kosukegawa, Hiroyuki
Abe, Toshihiko
Mori, Hitoshi
Mori, Kazumi
Takagi, Toshiyuki
Izumi, Shin-ichi
author_facet Kudo, Tada-aki
Kanetaka, Hiroyasu
Mochizuki, Kentaro
Tominami, Kanako
Nunome, Shoko
Abe, Genji
Kosukegawa, Hiroyuki
Abe, Toshihiko
Mori, Hitoshi
Mori, Kazumi
Takagi, Toshiyuki
Izumi, Shin-ichi
author_sort Kudo, Tada-aki
collection PubMed
description 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.
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spelling pubmed-43999382015-04-21 Induction of Neurite Outgrowth in PC12 Cells Treated with Temperature-Controlled Repeated Thermal Stimulation Kudo, Tada-aki Kanetaka, Hiroyasu Mochizuki, Kentaro Tominami, Kanako Nunome, Shoko Abe, Genji Kosukegawa, Hiroyuki Abe, Toshihiko Mori, Hitoshi Mori, Kazumi Takagi, Toshiyuki Izumi, Shin-ichi PLoS One Research Article 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. Public Library of Science 2015-04-16 /pmc/articles/PMC4399938/ /pubmed/25879210 http://dx.doi.org/10.1371/journal.pone.0124024 Text en © 2015 Kudo et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Kudo, Tada-aki
Kanetaka, Hiroyasu
Mochizuki, Kentaro
Tominami, Kanako
Nunome, Shoko
Abe, Genji
Kosukegawa, Hiroyuki
Abe, Toshihiko
Mori, Hitoshi
Mori, Kazumi
Takagi, Toshiyuki
Izumi, Shin-ichi
Induction of Neurite Outgrowth in PC12 Cells Treated with Temperature-Controlled Repeated Thermal Stimulation
title Induction of Neurite Outgrowth in PC12 Cells Treated with Temperature-Controlled Repeated Thermal Stimulation
title_full Induction of Neurite Outgrowth in PC12 Cells Treated with Temperature-Controlled Repeated Thermal Stimulation
title_fullStr Induction of Neurite Outgrowth in PC12 Cells Treated with Temperature-Controlled Repeated Thermal Stimulation
title_full_unstemmed Induction of Neurite Outgrowth in PC12 Cells Treated with Temperature-Controlled Repeated Thermal Stimulation
title_short Induction of Neurite Outgrowth in PC12 Cells Treated with Temperature-Controlled Repeated Thermal Stimulation
title_sort induction of neurite outgrowth in pc12 cells treated with temperature-controlled repeated thermal stimulation
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4399938/
https://www.ncbi.nlm.nih.gov/pubmed/25879210
http://dx.doi.org/10.1371/journal.pone.0124024
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