Yung-Hsiang Chen1,2,3, Chao-Jung Chen4,5, Yu-Ning Lin4, Yang-Chang Wu6, Wen-Tsong Hsieh4, Bor-Tsang Wu4, Wen-Lung Ma5, Wen-Chi Chen4,5, Kao-Sung Tsai4, San-Yuan Wu4,6, Chawnshang Chang5,7, Huey-Yi Chen8,9, Shuyuan Yeh10. 1. 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. 2. 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. 3. Department of Psychology, College of Medical and Health Science, Asia University, Taichung, 41354, Taiwan. d87a03@ym.edu.tw. 4. 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. 5. 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. 6. Graduate Institute of Natural Products, Kaohsiung Medical University, Kaohsiung, 80708, Taiwan. 7. Departments of Urology and Pathology, George H Whipple Laboratory for Cancer Research, Wilmot Cancer Center, University of Rochester Medical Center, Rochester, NY, 14642, USA. 8. 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. 9. 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. 10. 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.
Abstract
PURPOSE: 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.
PURPOSE: 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.
Entities:
Keywords:
Estrogen receptor α; Leak point pressures; Maximum urethral closure pressure; Proteomics; Stress urinary incontinence
Authors: Charuspong Dissaranan; Michelle A Cruz; Matthew J Kiedrowski; Brian M Balog; Bradley C Gill; Marc S Penn; Howard B Goldman; Margot S Damaser Journal: Cell Transplant Date: 2013-07-17 Impact factor: 4.064
Authors: Wilke M Post; Joanna Widomska; Hilde Grens; Marieke J H Coenen; Frank M J Martens; Dick A W Janssen; Joanna IntHout; Geert Poelmans; Egbert Oosterwijk; Kirsten B Kluivers Journal: Int J Mol Sci Date: 2022-03-21 Impact factor: 5.923