Thyroarytenoid Muscle Gene Expression in a Rat Model of Early‐Onset Parkinson's Disease

Abstract

Objectives/Hypothesis: Voice disorders in Parkinson's disease (PD) are early-onset, manifest in the preclinical stages of the disease, and negatively impact quality of life. The complete loss of function in the PTEN-induced kinase 1 gene (Pink1) causes a genetic form of early-onset, autosomal recessive PD. Modeled after the human inherited mutation, the Pink1À/À rat demonstrates significant cranial sensorimotor dysfunction including declines in ultrasonic vocalizations. However, the underlying genetics of the vocal fold thyroarytenoid (TA) muscle that may contribute to vocal deficits has not been studied. The aim of this study was to identify differentially expressed genes in the TA muscle of 8-month-old male Pink1À/À rats compared to wildtype controls.Study Design: Animal experiment with control. Methods: High throughput RNA sequencing was used to examine TA muscle gene expression in adult male Pink1À/À rats and wildtype controls. Weighted Gene Co-expression Network Analysis was used to construct co-expression modules to identify biological networks, including where Pink1 was a central node. The ENRICHR tool was used to compare this gene set to existing human gene databases.Results: We identified 134 annotated differentially expressed genes (P < .05 cutoff) and observed enrichment in the following biological pathways: Parkinson's disease (Casp7, Pink1); Parkin-Ubiquitin proteasome degradation (Psmd12, Psmd7); MAPK signaling (Casp7, Ppm1b, Ppp3r1); and inflammatory TNF-α, Nf-κB Signaling (Casp7, Psmd12, Psmd7, Cdc34, Bcl7a, Peg3).Conclusions: Genes and pathways identified here may be useful for evaluating the specific mechanisms of peripheral dysfunction including within the laryngeal muscle and have potential to be used as experimental biomarkers for treatment development.

PCBP-1 Regulates the Transcription and Alternative Splicing of Inflammation and Ubiquitination-Related Genes in PC12 Cell

“…Bcl7a has been described as B-cell CLL/lymphoma7 protein family member A, and it is involved in PD through the TNF-α and NF-κB signaling axis ( Wischhof et al, 2017 ). The genetic deletion of Bcl7a in postmitotic neurons elicits motor abnormalities ( Lechner et al, 2021 ). These findings provide novel proof for neuroinflammation and neuroimmune as the crucial modulators for PD pathophysiology.…”

An optimized method for high-quality RNA extraction from distinctive intrinsic laryngeal muscles in the rat model

“…A limiting factor of the preferred kit was its inability to consistently provide sufficient total RNA without pooling muscle fibers ( pt = 3). Compelling evidence in other rat model studies has demonstrated the feasibility of successful RNA-Seq analysis using the Aurum Total RNA Fatty and Fibrous Tissue Kit 25, 34 . Due to the greater column capacity (100 mg) of the aforementioned kit, its application can be assessed in future studies.…”

“…Rather than isolated ILM alone, they studied all the soft tissue associated with the VFs, including VF mucosa and respiratory epithelium. A recent Parkinson's study demonstrated RNA-Seq expression analysis from the rat TA; however, their sequencing strategy enabled the input of low-throughput RNA extracts 25 . This method, commonly known as TruSeq, requires a small sample size and has unique RNA specifications unamenable to other bulk RNA-Seq technologies 26 .…”

An optimized method for high-quality RNA extraction from distinctive intrinsic laryngeal muscles in the rat model

“…A limiting factor of the preferred kit was its inability to consistently provide su cient total RNA without pooling muscle bers (pt = 3). Compelling evidence in other rat model studies has demonstrated the feasibility of successful RNA-Seq analysis using the Aurum Total RNA Fatty and Fibrous Tissue Kit 25,34 . Due to the greater column capacity (100 mg) of the aforementioned kit, its application can be assessed in future studies.…”

“…Rather than isolated ILM alone, they studied all the soft tissue associated with the VFs, including VF mucosa and respiratory epithelium. A recent Parkinson's study demonstrated RNA-Seq expression analysis from the rat TA; however, their sequencing strategy enabled the input of low-throughput RNA extracts 25 . This method, commonly known as TruSeq, requires a small sample size and has unique RNA speci cations unamenable to other bulk RNA-Seq technologies 26 .…”