title: Proteomic analysis of urethral protein expression in an estrogen receptor α-deficient murine model of stress urinary incontinence.
abstract: OBJECTIVE
  The roles of estrogen receptor α (ERα) in stress urinary incontinence (SUI) remain elusive. This study was conducted to understand the molecular mechanism of ERα against SUI.
  
  
  METHODS
  Wild-type (ERα(+/+)) and ACTB-cre ERα knockout (ERα(-/-)) female mice were generated. Urethral function and protein expression were measured. Leak point pressures (LPP) and maximum urethral closure pressure (MUCP) were assessed in mice under urethane anesthesia. After the measurements, the urethras were removed for proteomic analysis using the two-dimensional differential gel electrophoresis and liquid chromatography-mass spectrometry technology. Interaction between these ERα pathway proteins was further analyzed by using MetaCore. Lastly, Western blot and immunochemistry (IHC) were used to confirm the candidate protein expression levels and locations, respectively.
  
  
  RESULTS
  Compared with the ERα(+/+) group, the LPP and MUCP values of the ERα(-/-) group were significantly decreased. Additionally, we identified 11 differentially expressed proteins in the urethra of ERα(-/-) female mice; five proteins were down-regulated and six were up-regulated. The majority of the ERα knockout-modified proteins were involved in muscle development, contraction, and regulation, as well as immune response (amphoterin signaling and phagocytosis), proteolysis, and cell adhesion (platelet aggregation and integrin-mediated cell-matrix adhesion). IHC and Western blot confirmed the down-regulation of tropomyosin and up-regulation of myosin in urethra.
  
  
  CONCLUSIONS
  This is the first study to estimate protein expression changes in urethras from ERα(-/-) female mice. These changes could be related to the molecular mechanism of ERα in SUI.
authors.affiliation: Graduate Institute of Integrated Medicine, School of Chinese Medicine, College of Chinese Medicine, School of Pharmacy, College of Pharmacy, Department of Pharmacology, Department of Physical Therapy, Graduate Institute of Rehabilitation Science, Research Center for Chinese Medicine and Acupuncture, China Medical University, Taichung, 40402, Taiwan. d87a03@ym.edu.tw.|Departments of Medical Research, Urology, and Obstetrics and Gynecology, Proteomics Core Laboratory, Sex Hormone Research Center, Chinese Medicine Research and Development Center, Center for Molecular Medicine, China Medical University Hospital, Taichung, 40402, Taiwan. d87a03@ym.edu.tw.|Department of Psychology, College of Medical and Health Science, Asia University, Taichung, 41354, Taiwan. d87a03@ym.edu.tw.
authors.forename: Yung-Hsiang
authors.initials: YH
authors.lastname: Chen
authors.affiliation: Graduate Institute of Integrated Medicine, School of Chinese Medicine, College of Chinese Medicine, School of Pharmacy, College of Pharmacy, Department of Pharmacology, Department of Physical Therapy, Graduate Institute of Rehabilitation Science, Research Center for Chinese Medicine and Acupuncture, China Medical University, Taichung, 40402, Taiwan.|Departments of Medical Research, Urology, and Obstetrics and Gynecology, Proteomics Core Laboratory, Sex Hormone Research Center, Chinese Medicine Research and Development Center, Center for Molecular Medicine, China Medical University Hospital, Taichung, 40402, Taiwan.
authors.forename: Chao-Jung
authors.initials: CJ
authors.lastname: Chen
authors.affiliation: Graduate Institute of Integrated Medicine, School of Chinese Medicine, College of Chinese Medicine, School of Pharmacy, College of Pharmacy, Department of Pharmacology, Department of Physical Therapy, Graduate Institute of Rehabilitation Science, Research Center for Chinese Medicine and Acupuncture, China Medical University, Taichung, 40402, Taiwan.
authors.forename: Yu-Ning
authors.initials: YN
authors.lastname: Lin
authors.affiliation: Graduate Institute of Natural Products, Kaohsiung Medical University, Kaohsiung, 80708, Taiwan.
authors.forename: Yang-Chang
authors.initials: YC
authors.lastname: Wu
authors.affiliation: Graduate Institute of Integrated Medicine, School of Chinese Medicine, College of Chinese Medicine, School of Pharmacy, College of Pharmacy, Department of Pharmacology, Department of Physical Therapy, Graduate Institute of Rehabilitation Science, Research Center for Chinese Medicine and Acupuncture, China Medical University, Taichung, 40402, Taiwan.
authors.forename: Wen-Tsong
authors.initials: WT
authors.lastname: Hsieh
authors.affiliation: Graduate Institute of Integrated Medicine, School of Chinese Medicine, College of Chinese Medicine, School of Pharmacy, College of Pharmacy, Department of Pharmacology, Department of Physical Therapy, Graduate Institute of Rehabilitation Science, Research Center for Chinese Medicine and Acupuncture, China Medical University, Taichung, 40402, Taiwan.
authors.forename: Bor-Tsang
authors.initials: BT
authors.lastname: Wu
authors.affiliation: Departments of Medical Research, Urology, and Obstetrics and Gynecology, Proteomics Core Laboratory, Sex Hormone Research Center, Chinese Medicine Research and Development Center, Center for Molecular Medicine, China Medical University Hospital, Taichung, 40402, Taiwan.
authors.forename: Wen-Lung
authors.initials: WL
authors.lastname: Ma
authors.affiliation: Graduate Institute of Integrated Medicine, School of Chinese Medicine, College of Chinese Medicine, School of Pharmacy, College of Pharmacy, Department of Pharmacology, Department of Physical Therapy, Graduate Institute of Rehabilitation Science, Research Center for Chinese Medicine and Acupuncture, China Medical University, Taichung, 40402, Taiwan.|Departments of Medical Research, Urology, and Obstetrics and Gynecology, Proteomics Core Laboratory, Sex Hormone Research Center, Chinese Medicine Research and Development Center, Center for Molecular Medicine, China Medical University Hospital, Taichung, 40402, Taiwan.
authors.forename: Wen-Chi
authors.initials: WC
authors.lastname: Chen
authors.affiliation: Graduate Institute of Integrated Medicine, School of Chinese Medicine, College of Chinese Medicine, School of Pharmacy, College of Pharmacy, Department of Pharmacology, Department of Physical Therapy, Graduate Institute of Rehabilitation Science, Research Center for Chinese Medicine and Acupuncture, China Medical University, Taichung, 40402, Taiwan.
authors.forename: Kao-Sung
authors.initials: KS
authors.lastname: Tsai
authors.affiliation: Graduate Institute of Integrated Medicine, School of Chinese Medicine, College of Chinese Medicine, School of Pharmacy, College of Pharmacy, Department of Pharmacology, Department of Physical Therapy, Graduate Institute of Rehabilitation Science, Research Center for Chinese Medicine and Acupuncture, China Medical University, Taichung, 40402, Taiwan.|Graduate Institute of Natural Products, Kaohsiung Medical University, Kaohsiung, 80708, Taiwan.
authors.forename: San-Yuan
authors.initials: SY
authors.lastname: Wu
authors.affiliation: Departments of Medical Research, Urology, and Obstetrics and Gynecology, Proteomics Core Laboratory, Sex Hormone Research Center, Chinese Medicine Research and Development Center, Center for Molecular Medicine, China Medical University Hospital, Taichung, 40402, Taiwan.|Departments of Urology and Pathology, George H Whipple Laboratory for Cancer Research, Wilmot Cancer Center, University of Rochester Medical Center, Rochester, NY, 14642, USA.
authors.forename: Chawnshang
authors.initials: C
authors.lastname: Chang
authors.affiliation: Graduate Institute of Integrated Medicine, School of Chinese Medicine, College of Chinese Medicine, School of Pharmacy, College of Pharmacy, Department of Pharmacology, Department of Physical Therapy, Graduate Institute of Rehabilitation Science, Research Center for Chinese Medicine and Acupuncture, China Medical University, Taichung, 40402, Taiwan. d888208@ms45.hinet.net.|Departments of Medical Research, Urology, and Obstetrics and Gynecology, Proteomics Core Laboratory, Sex Hormone Research Center, Chinese Medicine Research and Development Center, Center for Molecular Medicine, China Medical University Hospital, Taichung, 40402, Taiwan. d888208@ms45.hinet.net.
authors.forename: Huey-Yi
authors.initials: HY
authors.lastname: Chen
authors.affiliation: Departments of Urology and Pathology, George H Whipple Laboratory for Cancer Research, Wilmot Cancer Center, University of Rochester Medical Center, Rochester, NY, 14642, USA. shuyuan_yeh@urmc.rochester.edu.
authors.forename: Shuyuan
authors.initials: S
authors.lastname: Yeh
mesh_terms: D000818:Animals; D015153:Blotting, Western; D004195:Disease Models, Animal; D004586:Electrophoresis; D047628:Estrogen Receptor alpha / Q000096:biosynthesis / Q000172:deficiency / Q000235:genetics*; D005260:Female; D005786:Gene Expression Regulation; D005838:Genotype; D007150:Immunohistochemistry; D051379:Mice; D018345:Mice, Knockout; D016133:Polymerase Chain Reaction; D040901:Proteomics / Q000379:methods*; D012313:RNA / Q000235:genetics*; D014521:Urethra / Q000378:metabolism* / Q000473:pathology; D014550:Urinary Incontinence, Stress / Q000201:enzymology / Q000235:genetics*
chemical_list: D047628:Estrogen Receptor alpha; D012313:RNA
keywords: Estrogen receptor α; Leak point pressures; Maximum urethral closure pressure; Proteomics; Stress urinary incontinence