ABSTRACT
The lungs and the immune and nervous systems functionally interact to respond to respiratory environmental exposures and infections. The lungs are innervated by vagal sensory neurons of the jugular and nodose ganglia, fused together in smaller mammals as the jugular-nodose complex (JNC). Whereas the JNC shares properties with the other sensory ganglia, the trigeminal (TG) and dorsal root ganglia (DRG), these sensory structures express differential sets of genes that reflect their unique functionalities. Here, we used RNA sequencing (RNA-seq) in mice to identify the differential transcriptomes of the three sensory ganglia types. Using a fluorescent retrograde tracer and fluorescence-activated cell sorting, we isolated a defined population of airway-innervating JNC neurons and determined their differential transcriptional map after pulmonary exposure to lipopolysaccharide (LPS), a major mediator of acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) after infection with gram-negative bacteria or inhalation of organic dust. JNC neurons activated an injury response program, leading to increased expression of gene products such as the G protein-coupled receptor Cckbr, inducing functional changes in neuronal sensitivity to peptides, and Gpr151, also rapidly induced upon neuropathic nerve injury in pain models. Unique JNC-specific transcripts, present at only minimal levels in TG, DRG, and other organs, were identified. These included TMC3, encoding for a putative mechanosensor, and urotensin 2B, a hypertensive peptide. These findings highlight the unique properties of the JNC and reveal that ALI/ARDS rapidly induces a nerve injury-related state, changing vagal excitability.
Subject(s)
Nodose Ganglion/drug effects , Pneumonia/genetics , Receptor, Cholecystokinin B/genetics , Sensory Receptor Cells/drug effects , Transcriptome , Vagus Nerve Injuries/genetics , Animals , Ganglia, Spinal/drug effects , Ganglia, Spinal/immunology , Ganglia, Spinal/pathology , Gene Expression Profiling , Intracellular Signaling Peptides and Proteins/genetics , Intracellular Signaling Peptides and Proteins/immunology , Lipopolysaccharides/pharmacology , Lung/drug effects , Lung/immunology , Lung/pathology , Membrane Proteins/genetics , Membrane Proteins/immunology , Mice , Mice, Inbred C57BL , Nodose Ganglion/immunology , Nodose Ganglion/pathology , Peptide Hormones/genetics , Peptide Hormones/immunology , Pneumonia/chemically induced , Pneumonia/immunology , Pneumonia/pathology , Receptor, Cholecystokinin B/immunology , Receptors, G-Protein-Coupled/genetics , Receptors, G-Protein-Coupled/immunology , Sensory Receptor Cells/immunology , Sensory Receptor Cells/pathology , Sequence Analysis, RNA , Trigeminal Ganglion/drug effects , Trigeminal Ganglion/immunology , Trigeminal Ganglion/pathology , Vagus Nerve Injuries/chemically induced , Vagus Nerve Injuries/immunology , Vagus Nerve Injuries/pathologyABSTRACT
Laryngeal palsy often occurs as a result of recurrent laryngeal or vagal nerve injury during oncological surgery of the head and neck, affecting quality of life and increasing economic burden. Reinnervation following recurrent laryngeal nerve (RLN) injury is difficult despite development of techniques, such as neural anastomosis, nerve grafting and creation of a laryngeal muscle pedicle. In the present study, due to the limited availability of human nerve tissue for research, a rat model was used to investigate neurotrophin expression and laryngeal muscle pathophysiology in RLN injury. Twenty-five male Sprague-Dawley rats underwent right RLN transection with the excision of a 5-mm segment. Vocal fold movements, vocalization, histology and immunostaining were evaluated at different time-points (3, 6, 10 and 16 weeks). Although vocalization was restored, movement of the vocal fold failed to return to normal levels following RLN injury. The expression of brainderived neurotrophic factor and glial cell line-derived neurotrophic factor differed in the thyroarytenoid (TA) and posterior cricoarytenoid muscles. The number of axons did not increase to baseline levels over time. Furthermore, normal muscle function was unlikely with spontaneous reinnervation. During regeneration following RLN injury, differences in the expression levels of neurotrophic factors may have resulted in preferential reinnervation of the TA muscles. Data from the present study indicated that neurotrophic factors may be applied for restoring the function of the laryngeal nerve following recurrent injury.
Subject(s)
Laryngeal Nerve Injuries/physiopathology , Nerve Growth Factors/metabolism , Nerve Regeneration , Recurrent Laryngeal Nerve Injuries/drug therapy , Vagus Nerve Injuries/physiopathology , Animals , Axons/metabolism , Axons/pathology , Brain-Derived Neurotrophic Factor/metabolism , Glial Cell Line-Derived Neurotrophic Factor/metabolism , Humans , Laryngeal Muscles/drug effects , Laryngeal Muscles/metabolism , Laryngeal Muscles/physiopathology , Laryngeal Muscles/surgery , Laryngeal Nerve Injuries/drug therapy , Laryngeal Nerve Injuries/genetics , Laryngeal Nerve Injuries/surgery , Male , Neuroglia/drug effects , Neuroglia/pathology , Rats , Recurrent Laryngeal Nerve Injuries/genetics , Recurrent Laryngeal Nerve Injuries/physiopathology , Vagus Nerve Injuries/genetics , Vagus Nerve Injuries/surgery , Vocalization, Animal/drug effectsABSTRACT
OBJECTIVES: Recurrent laryngeal nerve (RLN) and vagus nerve (VN) injuries characteristically are followed by differing degrees of spontaneous reinnervation, yet laryngeal muscle neurotrophic factor (NF) expression profiles after RLN and VN injuries have not been well elucidated. This study's objective was to determine the relative changes in gene expression of 5 well-characterized NFs from laryngeal muscle after RLN or VN injuries in a time-dependent fashion, and demonstrate how these changes correspond with electromyography-assessed innervation status. METHODS: Thirty-six male rats underwent left RLN transection (12 rats), left VN transection (12 rats), or a sham procedure (12 rats). The primary outcomes included electromyographic assessment and laryngeal muscle NF expression quantification with reverse transcription polymerase chain reaction at 3 days and at 1 month. RESULTS: Electromyography at 3 days demonstrated electrical silence in the VN injury group, normal activity in the sham group, and nascent units with decreased recruitment in the RLN injury group. Reverse transcription polymerase chain reaction demonstrated that changes in NF gene expression from laryngeal muscles varied depending on the type of nerve injury (RLN or VN) and the specific laryngeal muscle (posterior cricoarytenoid or adductor) assessed. CONCLUSIONS: Laryngeal muscle NF expression profiles after cranial nerve X injury depend both upon the level of nerve injury and upon the muscles involved.