title: Molecular Assessment of Neuroregenerative Response in the Pudendal Nerve: A Useful Tool in Regenerative Urology.
abstract: OBJECTIVE
  Assessing pudendal nerve neuroregenerative response provides valuable insight into injuries and regenerative treatments related to urinary incontinence. This project developed and validated a cost-effective, expedient, and adoptable method of assessing pudendal nerve neuroregenerative response.
  
  
  METHODS
  Sprague Dawley rats underwent unilateral pudendal nerve crush prior to spinal cord harvest and laser microdissection for separate collection of the injured and uninjured Onuf's nuclei (pudendal motor neuron cell bodies). Commercially available kits were used to extract and isolate RNA, as well as reverse transcribe and amplify cDNA from cells. Utilizing standard quantitative polymerase chain reaction (Q-PCR), expression of βII-Tubulin, a cytoskeletal protein indicative of nerve growth and neuroregenerative response, was determined in the injured side relative to the uninjured side 1 week after injury.
  
  
  RESULTS
  Injury upregulated βII-Tubulin 2.36±0.46 times via Q-PCR, which was not significantly (p=0.508) different from the 2.49±0.38 times increase noted with in-situ hybridization previously. Starting with tissue collection, results are available within 1 day using PCR, while in-situ hybridization requires 4-weeks.
  
  
  CONCLUSIONS
  An easily adoptable PCR-based method of assessing the neuroregenerative response of the pudendal nerve successfully reproduced results obtained with a previous radioisotope-based in-situ hybridization technique.
authors.affiliation: Department of Urology, Glickman Urological and Kidney Institute, Cleveland Clinic, Cleveland, OH; Cleveland Clinic Lerner College of Medicine, Education Institute, Cleveland Clinic, Cleveland, OH; Department of Biomedical Engineering, Lerner Research Institute, Cleveland Clinic, Cleveland, OH.
authors.forename: Bradley C
authors.initials: BC
authors.lastname: Gill
authors.affiliation: Department of Biomedical Engineering, Lerner Research Institute, Cleveland Clinic, Cleveland, OH; Advanced Platform Technology Center, Louis Stokes Cleveland Veterans Affairs Medical Center, Cleveland, OH.
authors.forename: Dan Li
authors.initials: DL
authors.lastname: Lin
authors.affiliation: Department of Biomedical Engineering, Lerner Research Institute, Cleveland Clinic, Cleveland, OH; Advanced Platform Technology Center, Louis Stokes Cleveland Veterans Affairs Medical Center, Cleveland, OH.
authors.forename: Brian M
authors.initials: BM
authors.lastname: Balog
authors.affiliation: Department of Urology, Glickman Urological and Kidney Institute, Cleveland Clinic, Cleveland, OH; Department of Biomedical Engineering, Lerner Research Institute, Cleveland Clinic, Cleveland, OH.
authors.forename: Charuspong
authors.initials: C
authors.lastname: Dissaranan
authors.affiliation: Department of Urology, Glickman Urological and Kidney Institute, Cleveland Clinic, Cleveland, OH; Department of Biomedical Engineering, Lerner Research Institute, Cleveland Clinic, Cleveland, OH.
authors.forename: Hai-Hong
authors.initials: HH
authors.lastname: Jiang
authors.affiliation: Department of Urology, Glickman Urological and Kidney Institute, Cleveland Clinic, Cleveland, OH; Department of Biomedical Engineering, Lerner Research Institute, Cleveland Clinic, Cleveland, OH; Advanced Platform Technology Center, Louis Stokes Cleveland Veterans Affairs Medical Center, Cleveland, OH.
authors.forename: Margot S
authors.initials: MS
authors.lastname: Damaser
keywords: Beta-Tubulin; Nerve Regeneration; Neurogenic; Onuf’s Nucleus; Pudendal Nerve