Coordinated cadherin functions sculpt respiratory motor circuit connectivity

  1. Alicia N Vagnozzi
  2. Matthew T Moore
  3. Minshan Lin
  4. Elyse M Brozost
  5. Ritesh KC
  6. Aambar Agarwal
  7. Lindsay A Schwarz
  8. Xin Duan
  9. Niccolò Zampieri
  10. Lynn T Landmesser
  11. Polyxeni Philippidou  Is a corresponding author
  1. Case Western Reserve University, United States
  2. St. Jude Children's Research Hospital, United States
  3. University of California, San Francisco, United States
  4. Max Delbrück Center for Molecular Medicine, Germany

Abstract

Breathing, and the motor circuits that control it, are essential for life. At the core of respiratory circuits are Dbx1-derived interneurons, which generate the rhythm and pattern of breathing, and phrenic motor neurons (MNs), which provide the final motor output that drives diaphragm muscle contractions during inspiration. Despite their critical function, the principles that dictate how respiratory circuits assemble are unknown. Here we show that coordinated activity of a type I cadherin (N-cadherin) and type II cadherins (Cadherin-6, -9, and -10) is required in both MNs and Dbx1-derived neurons to generate robust respiratory motor output. Both MN- and Dbx1-specific cadherin inactivation in mice during a critical developmental window results in perinatal lethality due to respiratory failure and a striking reduction in phrenic MN bursting activity. This combinatorial cadherin code is required to establish phrenic MN cell body and dendritic topography; surprisingly, however, cell body position appears to be dispensable for the targeting of phrenic MNs by descending respiratory inputs. Our findings demonstrate that type I and type II cadherins function cooperatively throughout the respiratory circuit to generate a robust breathing output and reveal novel strategies that drive the assembly of motor circuits.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting files; source data files have been provided for Figures 1,2,3,4 and 6.

Article and author information

Author details

  1. Alicia N Vagnozzi

    Department of Neurosciences, Case Western Reserve University, Cleveland, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-6152-8728
  2. Matthew T Moore

    Department of Neurosciences, Case Western Reserve University, Cleveland, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Minshan Lin

    Department of Neurosciences, Case Western Reserve University, Cleveland, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Elyse M Brozost

    Department of Neurosciences, Case Western Reserve University, Cleveland, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Ritesh KC

    Department of Neurosciences, Case Western Reserve University, Cleveland, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Aambar Agarwal

    Department of Neurosciences, Case Western Reserve University, Cleveland, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Lindsay A Schwarz

    Department of Developmental Neurobiology, St. Jude Children's Research Hospital, Memphis, United States
    Competing interests
    The authors declare that no competing interests exist.
  8. Xin Duan

    Department of Ophthalmology, University of California, San Francisco, San Francisco, United States
    Competing interests
    The authors declare that no competing interests exist.
  9. Niccolò Zampieri

    Max Delbrück Center for Molecular Medicine, Berlin, Germany
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-2228-9453
  10. Lynn T Landmesser

    Department of Neurosciences, Case Western Reserve University, Cleveland, United States
    Competing interests
    The authors declare that no competing interests exist.
  11. Polyxeni Philippidou

    Department of Neurosciences, Case Western Reserve University, Cleveland, United States
    For correspondence
    pxp282@case.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-0733-3591

Funding

National Institute of Neurological Disorders and Stroke (R01NS114510)

  • Polyxeni Philippidou

National Eye Institute (R01EY030138)

  • Xin Duan

Eunice Kennedy Shriver National Institute of Child Health and Human Development (F30HD096788)

  • Alicia N Vagnozzi

National Institute of Neurological Disorders and Stroke (F31NS124240)

  • Matthew T Moore

National Institute of Neurological Disorders and Stroke (F31NS120699)

  • Ritesh KC

St. Jude Children's Research Hospital

  • Lindsay A Schwarz

NIHGM (T32GM007250)

  • Alicia N Vagnozzi

NIHGM (T32GM008056)

  • Ritesh KC

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Anne E West, Duke University, United States

Ethics

Animal experimentation: Mouse colony maintenance and handling was performed in compliance with protocols approved by the Institutional Animal Care Use Committee of Case Western Reserve University (assurance number: A-3145-01, protocol number: 2015-0180). Mice were housed in a 12-hour light/dark cycle in cages containing no more than five animals at a time.

Version history

  1. Received: July 23, 2022
  2. Preprint posted: August 4, 2022 (view preprint)
  3. Accepted: December 29, 2022
  4. Accepted Manuscript published: December 30, 2022 (version 1)
  5. Version of Record published: February 9, 2023 (version 2)

Copyright

© 2022, Vagnozzi et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 669
    views
  • 129
    downloads
  • 8
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Alicia N Vagnozzi
  2. Matthew T Moore
  3. Minshan Lin
  4. Elyse M Brozost
  5. Ritesh KC
  6. Aambar Agarwal
  7. Lindsay A Schwarz
  8. Xin Duan
  9. Niccolò Zampieri
  10. Lynn T Landmesser
  11. Polyxeni Philippidou
(2022)
Coordinated cadherin functions sculpt respiratory motor circuit connectivity
eLife 11:e82116.
https://doi.org/10.7554/eLife.82116

Share this article

https://doi.org/10.7554/eLife.82116

Further reading

    1. Developmental Biology
    Siyuan Cheng, Ivan Fan Xia ... Stefania Nicoli
    Research Article

    Vascular smooth muscle cells (VSMCs) envelop vertebrate brain arteries and play a crucial role in regulating cerebral blood flow and neurovascular coupling. The dedifferentiation of VSMCs is implicated in cerebrovascular disease and neurodegeneration. Despite its importance, the process of VSMC differentiation on brain arteries during development remains inadequately characterized. Understanding this process could aid in reprogramming and regenerating dedifferentiated VSMCs in cerebrovascular diseases. In this study, we investigated VSMC differentiation on zebrafish circle of Willis (CoW), comprising major arteries that supply blood to the vertebrate brain. We observed that arterial specification of CoW endothelial cells (ECs) occurs after their migration from cranial venous plexus to form CoW arteries. Subsequently, acta2+ VSMCs differentiate from pdgfrb+ mural cell progenitors after they were recruited to CoW arteries. The progression of VSMC differentiation exhibits a spatiotemporal pattern, advancing from anterior to posterior CoW arteries. Analysis of blood flow suggests that earlier VSMC differentiation in anterior CoW arteries correlates with higher red blood cell velocity and wall shear stress. Furthermore, pulsatile flow induces differentiation of human brain PDGFRB+ mural cells into VSMCs, and blood flow is required for VSMC differentiation on zebrafish CoW arteries. Consistently, flow-responsive transcription factor klf2a is activated in ECs of CoW arteries prior to VSMC differentiation, and klf2a knockdown delays VSMC differentiation on anterior CoW arteries. In summary, our findings highlight blood flow activation of endothelial klf2a as a mechanism regulating initial VSMC differentiation on vertebrate brain arteries.

    1. Developmental Biology
    Zhimin Xu, Zhao Wang ... Yingchuan B Qi
    Research Article

    Precise developmental timing control is essential for organism formation and function, but its mechanisms are unclear. In C. elegans, the microRNA lin-4 critically regulates developmental timing by post-transcriptionally downregulating the larval-stage-fate controller LIN-14. However, the mechanisms triggering the activation of lin-4 expression toward the end of the first larval stage remain unknown. We demonstrate that the transmembrane transcription factor MYRF-1 is necessary for lin-4 activation. MYRF-1 is initially localized on the cell membrane, and its increased cleavage and nuclear accumulation coincide with lin-4 expression timing. MYRF-1 regulates lin-4 expression cell-autonomously and hyperactive MYRF-1 can prematurely drive lin-4 expression in embryos and young first-stage larvae. The tandem lin-4 promoter DNA recruits MYRF-1GFP to form visible loci in the nucleus, suggesting that MYRF-1 directly binds to the lin-4 promoter. Our findings identify a crucial link in understanding developmental timing regulation and establish MYRF-1 as a key regulator of lin-4 expression.