Identification of lung innervating sensory neurons and their target specificity in mouse (3)

Study Purpose: Known as the gas exchange organ, the lung is also critical for responding to the aerosol environment in part through interaction with the nervous system. The diversity and specificity of lung innervating neurons remain poorly understood. Our goal wasto 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, 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 lung.

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=257) transgenic mice of unknown age 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. There is no derivative data folder.

Important notes: The dataset is undergoing image conversion to open format standards (JPEG2000 and OME-TIFF) and will be updated once this process is completed.