Identification of lung innervating sensory neurons and their target specificity in mouse (2)

Jamie Verheyden, Ph.D.
Xin Sun, Ph.D.

Immunostaining and imaging of Cre reporter lines in mouse lung lobes

Updated on October 4, 2023 (Version 1)

Corresponding Contributor:

Jamie Verheyden
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Dataset Overview

Study Purpose: To map the innervation pattern of the mouse lung and projections from the nodose ganglia.

Data Collection: This dataset contains Zeiss LightSheet immunofluorescence imaging of neuronal markers in mouse lung tissue.

Conclusions: During our SPARC project period we interrogated the cell body location, molecular signature, and projection pattern of lung innervating sensory neurons in the mouse. Retrograde tracing from the lung coupled with whole tissue clearing highlighted neurons primarily in the vagal ganglia. Centrally, they project specifically to the nucleus of the solitary tract in the brainstem. Peripherally, they enter the lung alongside branching airways. Labeling of nociceptor Trpv1+ versus peptidergic Tac1+ vagal neurons showed shared and distinct terminal morphology and targeting to airway smooth muscles, vasculature including lymphatics, and alveoli. Notably, a small population of Calb1+ neurons preferentially innervate pulmonary neuroendocrine cells, a demonstrated airway sensor population. This atlas of lung innervating neurons serves as a foundation for understanding their function in lungs.

Curator's Notes

Experimental Design: Mice were euthanized by CO2 inhalation then transcardially perfused with PBS followed by perfusion fixation with ice-cold 4% paraformaldehyde(PFA). The lungs were then inflated with 4% PFA and harvested. All the tissues were fixed overnight at 4C in 4% PFA. After fixing, tissues were washed three times in PBS. Tissues were then cleared by incubation in Clear, Unobstructed Brain/Body Imaging Cocktails and Computational analysis protocol (CUBIC) R1 solution on a rotating shaker (50 rpm) at 4C for 2–3 wk until visually cleared. For sectional analysis of fluorescent signals, tissues were washed in PBS to remove residual PFA and then cryoembedded in OCT compound following standard procedures. Blocks were sectioned at 40μm in rostral to caudal sequence and screened for fluorescence using a confocal microscope.

Completeness: This dataset is part of a larger study: "Foundational mapping of the neural circuits that control intrinsic lung function."

Subjects & Samples: Adult (n=51) transgenic mice between 8-12 weeks old were used in the study.

Primary vs derivative data: Primary data is organized by the subject ID and contains LightSheet fluorescence imaging of neuronal markers in mouse lung tissue. Derivative folder contains image data (JPEG2000 and OME-TIFF) that was derived from primary images (.CZI). .CZI images were converted with 20:1 compression to JPEG2000 (.jpx) by MBF Bioscience for web streaming and visualization on the SPARC Data Portal. .CZI images were also converted with lossless compression to OME-TIFF (.tif) by MBF Bioscience.


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Publishing history

October 4, 2023
Originally Published
October 4, 2023 (Version 1)
Last Updated

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