Go to JCI Insight
  • About
  • Editors
  • Consulting Editors
  • For authors
  • Publication ethics
  • Publication alerts by email
  • Advertising
  • Job board
  • Contact
  • Clinical Research and Public Health
  • Current issue
  • Past issues
  • By specialty
    • COVID-19
    • Cardiology
    • Gastroenterology
    • Immunology
    • Metabolism
    • Nephrology
    • Neuroscience
    • Oncology
    • Pulmonology
    • Vascular biology
    • All ...
  • Videos
    • Conversations with Giants in Medicine
    • Video Abstracts
  • Reviews
    • View all reviews ...
    • Complement Biology and Therapeutics (May 2025)
    • Evolving insights into MASLD and MASH pathogenesis and treatment (Apr 2025)
    • Microbiome in Health and Disease (Feb 2025)
    • Substance Use Disorders (Oct 2024)
    • Clonal Hematopoiesis (Oct 2024)
    • Sex Differences in Medicine (Sep 2024)
    • Vascular Malformations (Apr 2024)
    • View all review series ...
  • Viewpoint
  • Collections
    • In-Press Preview
    • Clinical Research and Public Health
    • Research Letters
    • Letters to the Editor
    • Editorials
    • Commentaries
    • Editor's notes
    • Reviews
    • Viewpoints
    • 100th anniversary
    • Top read articles

  • Current issue
  • Past issues
  • Specialties
  • Reviews
  • Review series
  • Conversations with Giants in Medicine
  • Video Abstracts
  • In-Press Preview
  • Clinical Research and Public Health
  • Research Letters
  • Letters to the Editor
  • Editorials
  • Commentaries
  • Editor's notes
  • Reviews
  • Viewpoints
  • 100th anniversary
  • Top read articles
  • About
  • Editors
  • Consulting Editors
  • For authors
  • Publication ethics
  • Publication alerts by email
  • Advertising
  • Job board
  • Contact
Selective antagonism of muscarinic receptors is neuroprotective in peripheral neuropathy
Nigel A. Calcutt, … , Corinne G. Jolivalt, Paul Fernyhough
Nigel A. Calcutt, … , Corinne G. Jolivalt, Paul Fernyhough
Published January 17, 2017
Citation Information: J Clin Invest. 2017;127(2):608-622. https://doi.org/10.1172/JCI88321.
View: Text | PDF
Research Article Neuroscience Article has an altmetric score of 133

Selective antagonism of muscarinic receptors is neuroprotective in peripheral neuropathy

  • Text
  • PDF
Abstract

Sensory neurons have the capacity to produce, release, and respond to acetylcholine (ACh), but the functional role of cholinergic systems in adult mammalian peripheral sensory nerves has not been established. Here, we have reported that neurite outgrowth from adult sensory neurons that were maintained under subsaturating neurotrophic factor conditions operates under cholinergic constraint that is mediated by muscarinic receptor–dependent regulation of mitochondrial function via AMPK. Sensory neurons from mice lacking the muscarinic ACh type 1 receptor (M1R) exhibited enhanced neurite outgrowth, confirming the role of M1R in tonic suppression of axonal plasticity. M1R-deficient mice made diabetic with streptozotocin were protected from physiological and structural indices of sensory neuropathy. Pharmacological blockade of M1R using specific or selective antagonists, pirenzepine, VU0255035, or muscarinic toxin 7 (MT7) activated AMPK and overcame diabetes-induced mitochondrial dysfunction in vitro and in vivo. These antimuscarinic drugs prevented or reversed indices of peripheral neuropathy, such as depletion of sensory nerve terminals, thermal hypoalgesia, and nerve conduction slowing in diverse rodent models of diabetes. Pirenzepine and MT7 also prevented peripheral neuropathy induced by the chemotherapeutic agents dichloroacetate and paclitaxel or HIV envelope protein gp120. As a variety of antimuscarinic drugs are approved for clinical use against other conditions, prompt translation of this therapeutic approach to clinical trials is feasible.

Authors

Nigel A. Calcutt, Darrell R. Smith, Katie Frizzi, Mohammad Golam Sabbir, Subir K. Roy Chowdhury, Teresa Mixcoatl-Zecuatl, Ali Saleh, Nabeel Muttalib, Randy Van der Ploeg, Joseline Ochoa, Allison Gopaul, Lori Tessler, Jürgen Wess, Corinne G. Jolivalt, Paul Fernyhough

×

Figure 2

Overexpression of GFP-M1R fusion protein inhibits neurite outgrowth.

Options: View larger image (or click on image) Download as PowerPoint
Overexpression of GFP-M1R fusion protein inhibits neurite outgrowth.
(A)...
(A) Immunoblot showing expression of GFP-tagged M1R protein in DRG neurons. M1R cDNA was cloned in PEGFP-C1 vector and used for transient transfection of DRG neurons using Amaxa Nucleofection reagent. Cells were harvested 24 hours after transfection, and proteins were separated by SDS-PAGE, followed by immunoblotting using anti-GFP and anti-M1R antibodies. (B) Bright-field (BF) and fluorescence images showing expression of GFP and GFP-M1R in neurons. Note extensive growth in GFP- vs. GFP-M1R–overexpressing neuron. (C) Fluorescent and immunostained images showing expression of GFP-tagged M1R in DRG neurons. Neurons were immunostained for β-tubulin III. Neurons with coexpression of GFP-M1R and β-tubulin III exhibited reduced neurite outgrowth compared with neurons expressing GFP alone. Scale bars: 100 μm. Colocalization of GFP (green) and β-tubulin III (red) indicated by yellow. (D) Neurons were transfected with GFP or GFP-M1R plasmids and maintained in vitro for 48 hours and immunostained for β-tubulin III. Total neurite outgrowth is shown as mean + SEM of n = 51 neurons; open circles indicate individual data points. ****P < 0.0001, Student’s unpaired t test. (E) Cultures overexpressing GFP-M1R were treated with 100 nM MT7 or 1 μM pirenzepine for 48 hours. Total neurite outgrowth is shown as group median with n = 100 neurons. ***P < 0.001; ****P < 0.0001 vs. control by 1-way ANOVA with Dunnett’s post-hoc test. Box and whisker plot where upper and lower limits of box indicate 75th and 25th percentiles, respectively. The middle lines show median, and error bars show maximum and minimum values. (F) WT (+/+) or M1R knockout (M1R-KO) mouse cultures were maintained for 48 hours in the presence of a low- (LD), medium- (MD), or high-dose (HD) neurotrophic factor cocktail. Total neurite outgrowth is shown as mean + SEM of n = 8 replicate cultures. *P < 0.05; **P < 0.01, Student’s t test.

Copyright © 2025 American Society for Clinical Investigation
ISSN: 0021-9738 (print), 1558-8238 (online)

Sign up for email alerts

Picked up by 15 news outlets
Blogged by 2
Posted by 24 X users
Referenced in 4 patents
On 1 Facebook pages
87 readers on Mendeley
See more details