Substance P Receptor in the Rat Heart and Regulation of Its Expression in Long-Term Diabetes

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Abstract

Substance P (SP) is a neuropeptide engaged in the signal transmission of neural C fibers afferents in the myocardium. The actions of SP in the heart are extensive and they are mediated by the neurokinin 1 receptor (NK1R), a member of the tachykinin subfamily of G-protein coupled receptors. The receptors have been found in the heart, but to our knowledge, their exact localization in the heart has not been described yet. Here, we investigated the presence of NK1R protein in separate rat heart compartments by means of western blot and its tissue distribution by means of immunofluorescence. Specificity of NK1R immunolabeling was controlled by preabsorption of the antiserum with its corresponding peptide. Additionally, we investigated abundance of gene for NK1R in separated heart chambers by means of quantitative real-time PCR (RT-PCR). Relative abundance of NK1R mRNA was expressed as a ratio of target gene Cq value to Cq value of control gene – beta-actin. Finally, we studied abundance of NK1R mRNA in different cell types of heart isolated by laser capture microdissection. Immunofluorescence showed NK1R immunoreactivity on the surface of some intracardiac neurons and smooth muscle cells of coronary vessels. The results of quantitative RT-PCR indicate abundance of mRNA for NK1R in all heart chambers with highest level in the left atrium. The presence of NK1R mRNA was detected in some samples of dissected intracardiac neurons, but not in cardiomyocytes or smooth muscle cells of coronary vessels. In the course of long-term diabetes, a significant downregulation of the NK1R mRNA was seen in the right atrium and upregulation in the right ventricle 53 weeks after the induction of diabetes. Our results indicate localization of NK1R in some intracardiac neurons and smooth muscle cells. Impaired transcription of the NK1R gene in the diabetic heart may be induced by unidentified genes or factors involved in the development of diabetic cardiomyopathy.

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PKC-1 regulates secretion of neuropeptides.

Jon Madison, Joshua Kaplan, Derek Sieburth (2006)

The secretion of neurotransmitters and neuropeptides is mediated by distinct organelles-synaptic vesicles (SVs) and dense-core vesicles (DCVs), respectively. Relatively little is known about the factors that differentially regulate SV and DCV secretion. Here we show that protein kinase C-1 (PKC-1), which is most similar to the vertebrate PKC eta and epsilon isoforms, regulates exocytosis of DCVs in Caenorhabditis elegans motor neurons. Mutants lacking PCK-1 activity had delayed paralysis induced by the acetylcholinesterase inhibitor aldicarb, whereas mutants with increased PKC-1 activity had more rapid aldicarb-induced paralysis. Imaging and electrophysiological assays indicated that SV release occurred normally in pkc-1 mutants. By contrast, genetic analysis of aldicarb responses and imaging of fluorescently tagged neuropeptides indicated that mutants lacking PKC-1 had reduced neuropeptide secretion. Similar neuropeptide secretion defects were found in mutants lacking unc-31 (encoding the protein CAPS) or unc-13 (encoding Munc13). These results suggest that PKC-1 selectively regulates DCV release from neurons.

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Human monocytes and macrophages express substance P and neurokinin-1 receptor.

W Z Ho, J P Lai, X H Zhu … (1997)

We present data demonstrating the gene expression of substance P and its receptor in human peripheral blood-isolated monocytes and macrophages. Using the RT-PCR assay, preprotachykinin-A (substance P) mRNA is detected in human peripheral blood-isolated monocytes and macrophages. Among the alpha, beta, and gamma transcripts of the substance P gene, only the beta and gamma transcripts are detectable in these cells. By Southern blot assay these RT-PCR-amplified transcripts are recognized using a specific substance P probe. Sequence analysis of the RT-PCR products from both monocytes and macrophages also confirmed the structure of these transcripts, which are identical to those found in human neuronal cells. At the protein level, both human monocytes and macrophages produced endogenous substance P as determined by an enzyme immunoassay. Capsaicin, a vanillyl fatty acid amide (ingredient of hot pepper), released substance P from both human monocytes and macrophages. In addition, using nested RT-PCR analysis, we identified the presence of mRNA for neurokinin-1 receptor (the receptor for substance P) in human peripheral blood-isolated monocytes and macrophages, which was confirmed by DNA sequencing analysis. The demonstration that human monocytes and macrophages express substance P and its receptor support the notion that substance P is biologically involved in regulating the functions of these cells in an autocrine fashion.

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Human lymphocytes express substance P and its receptor.

Jian-Ping Lai, Steven Douglas, Wen-Zhe Ho (1998)

We present data demonstrating the gene expression of substance P (SP) and its receptor in human peripheral blood-isolated lymphocytes. Using reverse transcribed polymerase chain reaction (RT-PCR) assay, preprotachykinin-A (substance-P) mRNA is detected in human peripheral blood-isolated lymphocytes. Among the alpha, beta, and gamma transcripts of the SP gene, only the beta and gamma transcripts are detectable in these cells. These RT-PCR amplified transcripts are recognized by Southern blot assay using a specific SP probe. Direct DNA sequence analysis of the RT-PCR products from lymphocytes also confirmed the structure of these transcripts which are identical to those found in human neuronal cells. At the protein level, human lymphocytes produced endogenous SP as determined by an enzyme immunoassay. Capsaicin, a vanillyl fatty acid amide (ingredient of hot pepper), released preformed SP from lymphocytes. In addition, using RT/nested-PCR analysis, we identified the presence of mRNA for neurokinin-1 receptor (the receptor for SP) in human peripheral blood-isolated lymphocytes, which was confirmed by Southern blot and DNA sequencing analysis. The demonstration that human lymphocytes express SP and its receptor support the notion that SP is biologically involved in regulating the functions of these cells in an autocrine fashion.

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Author and article information

Contributors

Magdalena Chottova Dvorakova: URI : http://loop.frontiersin.org/people/507182/overview

Renate Paddenberg: URI : http://loop.frontiersin.org/people/584022/overview

Wolfgang Kummer: URI : http://loop.frontiersin.org/people/109002/overview

Journal

Journal ID (nlm-ta): Front Physiol

Journal ID (iso-abbrev): Front Physiol

Journal ID (publisher-id): Front. Physiol.

Title: Frontiers in Physiology

Publisher: Frontiers Media S.A.

ISSN (Electronic): 1664-042X

Publication date (Electronic): 13 July 2018

Publication date Collection: 2018

Volume: 9

Electronic Location Identifier: 918

Affiliations

[1] ¹Biomedical Centre, Faculty of Medicine in Pilsen, Charles University , Pilsen, Czechia

[2] ²Department of Physiology, Faculty of Medicine in Pilsen, Charles University , Pilsen, Czechia

[3] ³Institute for Anatomy and Cell Biology, Justus-Liebig-University Giessen , Giessen, Germany

Author notes

Edited by: Scott Levick, The University of Sydney, Australia

Reviewed by: Steven Daniel Douglas, Children’s Hospital of Philadelphia, United States; Richard Barrett-Jolley, University of Liverpool, United Kingdom

*Correspondence: Magdalena Chottova Dvorakova, Magdalena.dvorakova@ 123456lfp.cuni.cz

This article was submitted to Integrative Physiology, a section of the journal Frontiers in Physiology

Article

DOI: 10.3389/fphys.2018.00918

PMC ID: 6053525

SO-VID: e01d3c04-8b87-49d1-b6da-a9e20ebc0b91

License:

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

History

Date received : 27 April 2018

Date accepted : 25 June 2018

Page count

Figures: 9, Tables: 1, Equations: 0, References: 54, Pages: 12, Words: 0

Funding

Funded by: Univerzita Karlova v Praze 10.13039/100007397

Award ID: Progres Q39

Funded by: Ministerstvo Školství, Mládeže a Tělovýchovy 10.13039/501100001823

Substance P Receptor in the Rat Heart and Regulation of Its Expression in Long-Term Diabetes

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Abstract

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In vitro alveolar region models

Most cited references 50

PKC-1 regulates secretion of neuropeptides.

Human monocytes and macrophages express substance P and neurokinin-1 receptor.

Human lymphocytes express substance P and its receptor.

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