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Differentiation dependent expression of TRPA1 and TRPM8 channels in IMR-32 human neuroblastoma cells.
J Cell Physiol. 2009 Oct; 221(1):67-74.JC

Abstract

TRPA1 and TRPM8 are transient receptor potential (TRP) channels involved in sensory perception. TRPA1 is a non-selective calcium permeable channel activated by irritants and proalgesic agents. TRPM8 reacts to chemical cooling agents such as menthol. The human neuroblastoma cell line IMR-32 undergoes a remarkable differentiation in response to treatment with 5-bromo-2-deoxyuridine. The cells acquire a neuronal morphology with increased expression of N-type voltage gated calcium channels and neurotransmitters. Here we show using RT-PCR, that mRNA for TRPA1 and TRPM8 are strongly upregulated in differentiating IMR-32 cells. Using whole cell patch clamp recordings, we demonstrate that activators of these channels, wasabi, allyl-isothiocyanate (AITC) and menthol activate membrane currents in differentiated cells. Calcium imaging experiments demonstrated that AITC mediated elevation of intracellular calcium levels were attenuated by ruthenium red, spermine, and HC-030031 as well as by siRNA directed against the channel. This indicates that the detected mRNA level correlate with the presence of functional channels of both types in the membrane of differentiated cells. Although the differentiated IMR-32 cells responded to cooling many of the cells showing this response did not respond to TRPA1/TRPM8 channel activators (60% and 90% for AITC and menthol respectively). Conversely many of the cells responding to these activators did not respond to cooling (30%). This suggests that these channels have also other functions than cold perception in these cells. Furthermore, our results suggest that IMR-32 cells have sensory characteristics and can be used to study native TRPA1 and TRPM8 channel function as well as developmental expression.

Authors+Show Affiliations

Biomedicum Helsinki, Institute of Biomedicine/Physiology, University of Helsinki, Helsinki, Finland.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

19507192

Citation

Louhivuori, Lauri M., et al. "Differentiation Dependent Expression of TRPA1 and TRPM8 Channels in IMR-32 Human Neuroblastoma Cells." Journal of Cellular Physiology, vol. 221, no. 1, 2009, pp. 67-74.
Louhivuori LM, Bart G, Larsson KP, et al. Differentiation dependent expression of TRPA1 and TRPM8 channels in IMR-32 human neuroblastoma cells. J Cell Physiol. 2009;221(1):67-74.
Louhivuori, L. M., Bart, G., Larsson, K. P., Louhivuori, V., Näsman, J., Nordström, T., Koivisto, A. P., & Akerman, K. E. (2009). Differentiation dependent expression of TRPA1 and TRPM8 channels in IMR-32 human neuroblastoma cells. Journal of Cellular Physiology, 221(1), 67-74. https://doi.org/10.1002/jcp.21828
Louhivuori LM, et al. Differentiation Dependent Expression of TRPA1 and TRPM8 Channels in IMR-32 Human Neuroblastoma Cells. J Cell Physiol. 2009;221(1):67-74. PubMed PMID: 19507192.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Differentiation dependent expression of TRPA1 and TRPM8 channels in IMR-32 human neuroblastoma cells. AU - Louhivuori,Lauri M, AU - Bart,Genevieve, AU - Larsson,Kim P, AU - Louhivuori,Verna, AU - Näsman,Johnny, AU - Nordström,Tommy, AU - Koivisto,Ari-Pekka, AU - Akerman,Karl E O, PY - 2009/6/10/entrez PY - 2009/6/10/pubmed PY - 2009/8/29/medline SP - 67 EP - 74 JF - Journal of cellular physiology JO - J Cell Physiol VL - 221 IS - 1 N2 - TRPA1 and TRPM8 are transient receptor potential (TRP) channels involved in sensory perception. TRPA1 is a non-selective calcium permeable channel activated by irritants and proalgesic agents. TRPM8 reacts to chemical cooling agents such as menthol. The human neuroblastoma cell line IMR-32 undergoes a remarkable differentiation in response to treatment with 5-bromo-2-deoxyuridine. The cells acquire a neuronal morphology with increased expression of N-type voltage gated calcium channels and neurotransmitters. Here we show using RT-PCR, that mRNA for TRPA1 and TRPM8 are strongly upregulated in differentiating IMR-32 cells. Using whole cell patch clamp recordings, we demonstrate that activators of these channels, wasabi, allyl-isothiocyanate (AITC) and menthol activate membrane currents in differentiated cells. Calcium imaging experiments demonstrated that AITC mediated elevation of intracellular calcium levels were attenuated by ruthenium red, spermine, and HC-030031 as well as by siRNA directed against the channel. This indicates that the detected mRNA level correlate with the presence of functional channels of both types in the membrane of differentiated cells. Although the differentiated IMR-32 cells responded to cooling many of the cells showing this response did not respond to TRPA1/TRPM8 channel activators (60% and 90% for AITC and menthol respectively). Conversely many of the cells responding to these activators did not respond to cooling (30%). This suggests that these channels have also other functions than cold perception in these cells. Furthermore, our results suggest that IMR-32 cells have sensory characteristics and can be used to study native TRPA1 and TRPM8 channel function as well as developmental expression. SN - 1097-4652 UR - https://www.unboundmedicine.com/medline/citation/19507192/Differentiation_dependent_expression_of_TRPA1_and_TRPM8_channels_in_IMR_32_human_neuroblastoma_cells_ L2 - https://doi.org/10.1002/jcp.21828 DB - PRIME DP - Unbound Medicine ER -