Cargando…

Functional Expression of TRPV1 Ion Channel in the Canine Peripheral Blood Mononuclear Cells

TRPV1, known as a capsaicin receptor, is the best-described transient receptor potential (TRP) ion channel. Recently, it was shown to be expressed by non-excitable cells such as lymphocytes. However, the data regarding the functional expression of the TRPV1 channel in the immune cells are often cont...

Descripción completa

Detalles Bibliográficos
Autores principales: Bujak, Joanna K., Kosmala, Daria, Majchrzak-Kuligowska, Kinga, Bednarczyk, Piotr
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8003907/
https://www.ncbi.nlm.nih.gov/pubmed/33804707
http://dx.doi.org/10.3390/ijms22063177
_version_ 1783671799963189248
author Bujak, Joanna K.
Kosmala, Daria
Majchrzak-Kuligowska, Kinga
Bednarczyk, Piotr
author_facet Bujak, Joanna K.
Kosmala, Daria
Majchrzak-Kuligowska, Kinga
Bednarczyk, Piotr
author_sort Bujak, Joanna K.
collection PubMed
description TRPV1, known as a capsaicin receptor, is the best-described transient receptor potential (TRP) ion channel. Recently, it was shown to be expressed by non-excitable cells such as lymphocytes. However, the data regarding the functional expression of the TRPV1 channel in the immune cells are often contradictory. In the present study, we performed a phylogenetical analysis of the canine TRP ion channels, we assessed the expression of TRPV1 in the canine peripheral blood mononuclear cells (PBMC) by qPCR and Western blot, and we determined the functionality of TRPV1 by whole-cell patch-clamp recordings and calcium assay. We found high expression of TRPV2, -M2, and -M7 in the canine PBMCs, while expression of TRPV1, -V4 and, -M5 was relatively low. We confirmed that TRPV1 is expressed on the protein level in the PBMC and it localizes in the plasma membrane. The whole-cell patch-clamp recording revealed that capsaicin application caused a significant increase in the current density. Similarly, the results from the calcium assay show a dose-dependent increase in intracellular calcium level in the presence of capsaicin that was partially abolished by capsazepine. Our study confirms the expression of TRPV1 ion channel on both mRNA and protein levels in the canine PBMC and indicates that the ion channel is functional.
format Online
Article
Text
id pubmed-8003907
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-80039072021-03-28 Functional Expression of TRPV1 Ion Channel in the Canine Peripheral Blood Mononuclear Cells Bujak, Joanna K. Kosmala, Daria Majchrzak-Kuligowska, Kinga Bednarczyk, Piotr Int J Mol Sci Article TRPV1, known as a capsaicin receptor, is the best-described transient receptor potential (TRP) ion channel. Recently, it was shown to be expressed by non-excitable cells such as lymphocytes. However, the data regarding the functional expression of the TRPV1 channel in the immune cells are often contradictory. In the present study, we performed a phylogenetical analysis of the canine TRP ion channels, we assessed the expression of TRPV1 in the canine peripheral blood mononuclear cells (PBMC) by qPCR and Western blot, and we determined the functionality of TRPV1 by whole-cell patch-clamp recordings and calcium assay. We found high expression of TRPV2, -M2, and -M7 in the canine PBMCs, while expression of TRPV1, -V4 and, -M5 was relatively low. We confirmed that TRPV1 is expressed on the protein level in the PBMC and it localizes in the plasma membrane. The whole-cell patch-clamp recording revealed that capsaicin application caused a significant increase in the current density. Similarly, the results from the calcium assay show a dose-dependent increase in intracellular calcium level in the presence of capsaicin that was partially abolished by capsazepine. Our study confirms the expression of TRPV1 ion channel on both mRNA and protein levels in the canine PBMC and indicates that the ion channel is functional. MDPI 2021-03-20 /pmc/articles/PMC8003907/ /pubmed/33804707 http://dx.doi.org/10.3390/ijms22063177 Text en © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Bujak, Joanna K.
Kosmala, Daria
Majchrzak-Kuligowska, Kinga
Bednarczyk, Piotr
Functional Expression of TRPV1 Ion Channel in the Canine Peripheral Blood Mononuclear Cells
title Functional Expression of TRPV1 Ion Channel in the Canine Peripheral Blood Mononuclear Cells
title_full Functional Expression of TRPV1 Ion Channel in the Canine Peripheral Blood Mononuclear Cells
title_fullStr Functional Expression of TRPV1 Ion Channel in the Canine Peripheral Blood Mononuclear Cells
title_full_unstemmed Functional Expression of TRPV1 Ion Channel in the Canine Peripheral Blood Mononuclear Cells
title_short Functional Expression of TRPV1 Ion Channel in the Canine Peripheral Blood Mononuclear Cells
title_sort functional expression of trpv1 ion channel in the canine peripheral blood mononuclear cells
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8003907/
https://www.ncbi.nlm.nih.gov/pubmed/33804707
http://dx.doi.org/10.3390/ijms22063177
work_keys_str_mv AT bujakjoannak functionalexpressionoftrpv1ionchannelinthecanineperipheralbloodmononuclearcells
AT kosmaladaria functionalexpressionoftrpv1ionchannelinthecanineperipheralbloodmononuclearcells
AT majchrzakkuligowskakinga functionalexpressionoftrpv1ionchannelinthecanineperipheralbloodmononuclearcells
AT bednarczykpiotr functionalexpressionoftrpv1ionchannelinthecanineperipheralbloodmononuclearcells