Skip to main content
Advertisement

Main menu

  • Home
  • Articles
    • Current Issue
    • Fast Forward
    • Latest Articles
    • Special Sections
    • Archive
  • Information
    • Instructions to Authors
    • Submit a Manuscript
    • FAQs
    • For Subscribers
    • Terms & Conditions of Use
    • Permissions
  • Editorial Board
  • Alerts
    • Alerts
    • RSS Feeds
  • Virtual Issues
  • Feedback
  • Submit
  • Other Publications
    • Drug Metabolism and Disposition
    • Journal of Pharmacology and Experimental Therapeutics
    • Molecular Pharmacology
    • Pharmacological Reviews
    • Pharmacology Research & Perspectives
    • ASPET

User menu

  • My alerts
  • Log in
  • My Cart

Search

  • Advanced search
Journal of Pharmacology and Experimental Therapeutics
  • Other Publications
    • Drug Metabolism and Disposition
    • Journal of Pharmacology and Experimental Therapeutics
    • Molecular Pharmacology
    • Pharmacological Reviews
    • Pharmacology Research & Perspectives
    • ASPET
  • My alerts
  • Log in
  • My Cart
Journal of Pharmacology and Experimental Therapeutics

Advanced Search

  • Home
  • Articles
    • Current Issue
    • Fast Forward
    • Latest Articles
    • Special Sections
    • Archive
  • Information
    • Instructions to Authors
    • Submit a Manuscript
    • FAQs
    • For Subscribers
    • Terms & Conditions of Use
    • Permissions
  • Editorial Board
  • Alerts
    • Alerts
    • RSS Feeds
  • Virtual Issues
  • Feedback
  • Submit
  • Visit jpet on Facebook
  • Follow jpet on Twitter
  • Follow jpet on LinkedIn
Research ArticleGastrointestinal, Hepatic, Pulmonary, and Renal
Open Access

Gender-Related Differences of Tachykinin NK2 Receptor Expression and Activity in Human Colonic Smooth Muscle

Stelina Drimousis, Irit Markus, Tim V. Murphy, D. Shevy Perera, Kim-Chi Phan-Thien, Li Zhang and Lu Liu
Journal of Pharmacology and Experimental Therapeutics October 2020, 375 (1) 28-39; DOI: https://doi.org/10.1124/jpet.120.265967
Stelina Drimousis
Department of Pharmacology (S.D., I.M., L.L.), Department of Physiology (T.V.M.), School of Medical Sciences, UNSW Sydney, New South Wales, Australia; Sydney Colorectal Associates, Hurstville, New South Wales, Australia (D.S.P., K.-C.P.-T.); and School of Biotechnology and Biomolecular Sciences, UNSW Sydney, New South Wales, Australia (L.Z.)
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Irit Markus
Department of Pharmacology (S.D., I.M., L.L.), Department of Physiology (T.V.M.), School of Medical Sciences, UNSW Sydney, New South Wales, Australia; Sydney Colorectal Associates, Hurstville, New South Wales, Australia (D.S.P., K.-C.P.-T.); and School of Biotechnology and Biomolecular Sciences, UNSW Sydney, New South Wales, Australia (L.Z.)
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Tim V. Murphy
Department of Pharmacology (S.D., I.M., L.L.), Department of Physiology (T.V.M.), School of Medical Sciences, UNSW Sydney, New South Wales, Australia; Sydney Colorectal Associates, Hurstville, New South Wales, Australia (D.S.P., K.-C.P.-T.); and School of Biotechnology and Biomolecular Sciences, UNSW Sydney, New South Wales, Australia (L.Z.)
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
D. Shevy Perera
Department of Pharmacology (S.D., I.M., L.L.), Department of Physiology (T.V.M.), School of Medical Sciences, UNSW Sydney, New South Wales, Australia; Sydney Colorectal Associates, Hurstville, New South Wales, Australia (D.S.P., K.-C.P.-T.); and School of Biotechnology and Biomolecular Sciences, UNSW Sydney, New South Wales, Australia (L.Z.)
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Kim-Chi Phan-Thien
Department of Pharmacology (S.D., I.M., L.L.), Department of Physiology (T.V.M.), School of Medical Sciences, UNSW Sydney, New South Wales, Australia; Sydney Colorectal Associates, Hurstville, New South Wales, Australia (D.S.P., K.-C.P.-T.); and School of Biotechnology and Biomolecular Sciences, UNSW Sydney, New South Wales, Australia (L.Z.)
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Li Zhang
Department of Pharmacology (S.D., I.M., L.L.), Department of Physiology (T.V.M.), School of Medical Sciences, UNSW Sydney, New South Wales, Australia; Sydney Colorectal Associates, Hurstville, New South Wales, Australia (D.S.P., K.-C.P.-T.); and School of Biotechnology and Biomolecular Sciences, UNSW Sydney, New South Wales, Australia (L.Z.)
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Lu Liu
Department of Pharmacology (S.D., I.M., L.L.), Department of Physiology (T.V.M.), School of Medical Sciences, UNSW Sydney, New South Wales, Australia; Sydney Colorectal Associates, Hurstville, New South Wales, Australia (D.S.P., K.-C.P.-T.); and School of Biotechnology and Biomolecular Sciences, UNSW Sydney, New South Wales, Australia (L.Z.)
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • Article
  • Figures & Data
  • Info & Metrics
  • eLetters
  • PDF + SI
  • PDF
Loading

Article Figures & Data

Figures

  • Tables
  • Additional Files
  • Fig. 1.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 1.

    Gender-related differences in NK2 receptor–mediated contractility. Contractile responses of female (n = 11) and male (n = 10 to 11) human colonic circular smooth muscle strips to (A) the endogenous NK2 receptor ligand NKA and (B) the selective NK2 receptor agonist LMN-NKA. Data are expressed as mean tension (grams) ± S.E.M. Significance by two-way ANOVA is indicated.

  • Fig. 2.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 2.

    Gender-specific distinctions in the potency of the selective NK2 receptor antagonist ibodutant. The effect of different concentrations of ibodutant on contractile responses induced by the endogenous NK2 receptor ligand NKA (left panel) and the selective NK2 receptor agonist LMN-NKA (right panel) in female (A and B) and male (C and D) human colonic circular smooth muscle strips. Data are expressed as a tension (grams), and curves are presented as a percentage of the mean maximum response for each concentration of ibodutant. n = 9–11 for each curve, and closed circles indicate means ± S.E.M.

  • Fig. 3.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 3.

    The Schild plots generated to calculate pA2 values for ibodutant against the endogenous NK2 receptor ligand NKA (left panel) and the selective NK2 receptor agonist LMN-NKA (right panel) in female (A and B) and male (C and D) human colonic circular muscle strips (n = 11).

  • Fig. 4.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 4.

    Gender-related differences in the neuronal release of tachykinin peptides. (A and B) Basal release of NKA and SP from female (n = 14 and 10, respectively) and male (n = 15 and nine, respectively) human colonic smooth muscle strips. Horizontal lines denote medians. Significance by Mann-Whitney test is indicated, where *P = 0.0279. (C and D) The release of NKA and SP in response to increasing frequencies of EFS. Data are expressed as a percentage of the basal amount released and are presented as means ± S.E.M. One-sample t test analysis of EFS-induced tachykinin release compared with basal level is indicated by *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.

  • Fig. 5.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 5.

    Western blot and densitometry analysis of NK2 receptor protein expression in human colonic sigmoid smooth muscle. Results are expressed as a percentage of the loading control GAPDH. (A) NK2 receptor and GAPDH protein blots with specific bands corresponding to the expected molecular weight of ∼45 and ∼36 kDa, respectively. (B) NK2 receptor protein expression in female (n = 25) and male (n = 15) samples. Horizontal lines denote medians. (C) NK2 receptor protein expression in samples from patients younger (closed circles) or older (open circles) than 50 years of age. (D) Spearman correlation analysis of NK2 receptor expression and age in females (r = 0.5023, *P = 0.0105) and males (r = 0.127, P = 0.6499).

  • Fig. 6.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 6.

    Fluorescent immunoreactivity of the NK2 receptor (left panel) in the smooth muscle layers of the human sigmoid colon. The cell markers used (middle panel) are α-SMA (B, n = 2), the interstitial cells of Cajal marker c-kit (E and H, n = 2), and the neuronal marker β-tubulin (K, n = 6). The merged images (right panel) contain the nuclear marker DAPI (blue). Staining is present on myenteric ganglia (mg), circular muscle (cm), and longitudinal (lm) muscle. Scale bar, 20 µm. Representative images were taken from female specimens aged 62 years old (A–C and J–L) and 46 years old (G–I) and a male specimen aged 49 years old (D–F).

  • Fig. 7.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 7.

    Fluorescent staining of the NK2 receptor (left panel) within the myenteric ganglia of the human sigmoid colon. The cell markers used (middle panel) are the nerve cell body marker HuC/D (B, n = 2), the neuronal marker β-tubulin (E, n = 6), and the glial cell markers S100 (H, n = 6) and GFAP (K, n = 2). The merged images (right panel) contain the nuclear marker DAPI (blue). Staining is present on myenteric ganglia (mg), circular muscle (cm), and longitudinal (lm) muscle. Scale bar, 20 µm. Representative images were taken from a female specimen aged 62 years old (A–F) and a male specimen aged 49 years old (G–L).

  • Fig. 8.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 8.

    Gender-related variations in NK2 receptor–mediated second messenger signaling pathways. Contractile responses of female (red) and male (blue) human colonic circular smooth muscle strips to NKA (left panel) and LMN-NKA (right panel) at 0.3 μM, either alone (dashed lines) or in the presence of the PLC inhibitor U73122 (A and B), the inositol IP3 receptor inhibitor 2-APB (C and D), the PKC inhibitor GF109203X (E and F), or the Rho kinase inhibitor Y-27632 dihydrochloride (G and H) at 3, 30, 100, and 300 μM. Data are expressed as a percentage of the contractile response before pharmacological inhibition and are presented as means ± S.E.M. (n = 5–11 for each curve). Significance by two-way ANOVA is indicated by a single symbol for P < 0.05, two for P < 0.01, three for P < 0.001, and four for P < 0.0001 (* for before vs. after inhibition within each gender; † for females vs. males).

  • Fig. 9.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 9.

    Gene expression of the Gαq family of G proteins (A–C) and the PLCβ isoenzymes (D–G) in female (n = 26 to 27) and male (n = 25–27) human colonic smooth muscle. Results are normalized to GAPDH expression and presented as fold change relative to the inter-run calibrator. Horizontal lines denote medians.

Tables

  • Figures
  • Additional Files
    • View popup
    TABLE 1

    Summary of Schild plot results for ibodutant against NKA and the selective NK2 receptor agonist LMN-NKA in human colonic circular smooth muscle

    pA2 values and slopes were generated from Schild plot and expressed as means ± S.E.M. All slopes were not significantly different from 1, and there was no difference between the slopes. Values in parentheses are the mean apparent pKB of individual ibodutant concentrations calculated using the equation pKB = log (dose ratio − 1) − log [B], where [B] is the antagonist concentration (Arunlakshana and Schild, 1959; Kenakin, 2009).

    ParametersIbodutant against NKAIbodutant against LMN-NKA
    FemaleMaleFemaleMale
    pA2 ± S.E.M. (apparent pKB)8.36 ± 0.38* (8.22 ± 0.17)**7.26 ± 0.21 (7.10 ± 0.14)8.14 ± 0.36 (7.99 ± 0.19)8.33 ± 0.22* (8.11 ± 0.08)**
    Schild slope ± S.E.M.0.76 ± 0.140.81 ± 0.0550.72 ± 0.170.90 ± 0.12
    Slope different from 1NoNoNoNo
    P = 0.24P = 0.077P = 0.24P = 0.51
    • ↵* P < 0.05; **P < 0.01, compared with male against NKA (one-way ANOVA with Bonferroni’s test).

Additional Files

  • Figures
  • Tables
  • Data Supplement

    • Supplemental Data -

      Supplementary Table 1 - Forward (FP) and reverse (RP) primers of human target and housekeeping genes used in real-time PCR studies. Primer melting temperatures and amplification efficiency scores are shown.

      Supplementary Figure 1 - Contractile responses of human colonic circular muscle to A) the endogenous NK2 receptor ligand neurokinin A (NKA), B) the selective NK2 receptor agonist [Lys5,MeLeu9, Nle10]NKA(4-10) (LMN-NKA) and C) the selective NK1 receptor agonist [Pro9] substance P ([Pro9] SP). 

      Supplementary Figure 2 - The effect of atropine on the contractile responses of human colonic circular muscle to A) the endogenous NK2 receptor ligand neurokinin A (NKA) and B) the selective NK2 receptor agonist [Lys5
      ,MeLeu9, Nle10]NKA(4-10) (LMN-NKA).

PreviousNext
Back to top

In this issue

Journal of Pharmacology and Experimental Therapeutics: 375 (1)
Journal of Pharmacology and Experimental Therapeutics
Vol. 375, Issue 1
1 Oct 2020
  • Table of Contents
  • Table of Contents (PDF)
  • About the Cover
  • Index by author
  • Editorial Board (PDF)
  • Front Matter (PDF)
Download PDF
Article Alerts
Sign In to Email Alerts with your Email Address
Email Article

Thank you for sharing this Journal of Pharmacology and Experimental Therapeutics article.

NOTE: We request your email address only to inform the recipient that it was you who recommended this article, and that it is not junk mail. We do not retain these email addresses.

Enter multiple addresses on separate lines or separate them with commas.
Gender-Related Differences of Tachykinin NK2 Receptor Expression and Activity in Human Colonic Smooth Muscle
(Your Name) has forwarded a page to you from Journal of Pharmacology and Experimental Therapeutics
(Your Name) thought you would be interested in this article in Journal of Pharmacology and Experimental Therapeutics.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Citation Tools
Research ArticleGastrointestinal, Hepatic, Pulmonary, and Renal

Tachykinin NK2 Receptors in Human Colonic Smooth Muscle

Stelina Drimousis, Irit Markus, Tim V. Murphy, D. Shevy Perera, Kim-Chi Phan-Thien, Li Zhang and Lu Liu
Journal of Pharmacology and Experimental Therapeutics October 1, 2020, 375 (1) 28-39; DOI: https://doi.org/10.1124/jpet.120.265967

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero

Share
Research ArticleGastrointestinal, Hepatic, Pulmonary, and Renal

Tachykinin NK2 Receptors in Human Colonic Smooth Muscle

Stelina Drimousis, Irit Markus, Tim V. Murphy, D. Shevy Perera, Kim-Chi Phan-Thien, Li Zhang and Lu Liu
Journal of Pharmacology and Experimental Therapeutics October 1, 2020, 375 (1) 28-39; DOI: https://doi.org/10.1124/jpet.120.265967
del.icio.us logo Digg logo Reddit logo Twitter logo Facebook logo Google logo Mendeley logo
  • Tweet Widget
  • Facebook Like
  • Google Plus One

Jump to section

  • Article
    • Abstract
    • Introduction
    • Materials and Methods
    • Results
    • Discussion
    • Acknowledgments
    • Authorship Contributions
    • Footnotes
    • Abbreviations
    • References
  • Figures & Data
  • Info & Metrics
  • eLetters
  • PDF + SI
  • PDF

Related Articles

Cited By...

More in this TOC Section

  • A Novel Long-Acting GLP-2, HM15912, for Short Bowel Syndrome
  • H2S Overproduction and Colonic Hypomotility in DM
  • MIP3a in Progressive Renal Injury Associated With Obesity
Show more Gastrointestinal, Hepatic, Pulmonary, and Renal

Similar Articles

Advertisement
  • Home
  • Alerts
Facebook   Twitter   LinkedIn   RSS

Navigate

  • Current Issue
  • Fast Forward by date
  • Fast Forward by section
  • Latest Articles
  • Archive
  • Search for Articles
  • Feedback
  • ASPET

More Information

  • About JPET
  • Editorial Board
  • Instructions to Authors
  • Submit a Manuscript
  • Customized Alerts
  • RSS Feeds
  • Subscriptions
  • Permissions
  • Terms & Conditions of Use

ASPET's Other Journals

  • Drug Metabolism and Disposition
  • Molecular Pharmacology
  • Pharmacological Reviews
  • Pharmacology Research & Perspectives
ISSN 1521-0103 (Online)

Copyright © 2023 by the American Society for Pharmacology and Experimental Therapeutics