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    Korean J Urol. 2009 May;50(5):498-504. Korean.
    Published online May 26, 2009.
    https://doi.org/10.4111/kju.2009.50.5.498
    Copyright © The Korean Urological Association, 2009
    Original Article

    Proteomic Analysis of Penile Protein Alterations in a Rat Model of Cavernous Nerve Injury

    Hong Chung, Chang-Kwon Lee,1 Bokyung Kim,1 Hong Sup Kim, Tong-Wook Kim, Sung Hyun Paick, Hyun Soo Jeon,2 and Sang-Kuk Yang
      • Department of Urology, College of Medicine, Konkuk University, Chungju, Korea.
      • 1Department of Physiology, College of Medicine, Konkuk University, Chungju, Korea.
      • 2Department of Obstetrics and Gynecology, College of Medicine, Konkuk University, Chungju, Korea.
    • Correspondence to: Sang-Kuk Yang. Department of Urology, College of Medicine, Konkuk University, 620-5, Kyohyun 2-dong, Chungju 380-704, Korea. TEL: 043-840-8270, FAX: 043-848-4722, Email: yskurol@kku.ac.kr
    Received March 05, 2009; Accepted April 22, 2009.

    Abstract

    Purpose

    Cavernous nerve resection (CNR) in rats is a standard model of animal experiments on erectile dysfunction (ED) that occurs after radical prostatectomy (RP). Injured cavernous nerves after surgery can cause fibrosis and apoptosis that lead to penile structural changes that may be accompanied by alterations of protein expression. This study aimed to analyze the changes in protein after CNR in Wistar Kyoto rats.

    Materials and Methods

    Using 8-week-old male Wistar Kyoto rats, sham and CNR operation under a microscope were performed. Two and 8 weeks after surgery, we applied 2-DE and MALDI-TOF/TOF (AB 4700) to identify differently expressed penile proteins after CNR. 2-DE gels were stained with silver nitrate and were analyzed with PDQuest. After in-gel digestion, peptide mass spectra were obtained by MALDI-TOF/TOF mass spectrometry in the positive ion reflector mode. The obtained data were screened with a rat database from both the NCBI and the Swiss-Prot/TrFMBL home page.

    Results

    The proteins that were changed more than 1.5-fold compared with the sham group were annexin A4 and pyruvate kinase (PK). Annexin A4 was increased by 1.75-fold after 2 weeks, whereas PK was decreased by 4.16 after 8 weeks. These results were confirmed by immunohistochemistry.

    Conclusions

    Annexin A4 in the CNR group was increased, which may be related to emiocytosis during apoptosis. The decrease in PK of the CNR group is assumed to be related to a decrease in efficacy during glycolysis. Further study will be needed to elucidate the molecular pathophysiology of ED after cavernous nerve injury.

    Keywords
    Cavernous nerve; Erectile dysfunction; Proteomics

    Figures

    Fig. 1
    Two-dimension electrophoresis image of the rat penile cavernosum tissue. (A) after 2 weeks, (B) after 8 weeks (1: Annexin A4, 2: Pyruvate kinase, WKY: Wistar Kyoto rats, CNR: cavernous nerves resection).

    Fig. 2
    Two-dimension electrophoresis patterns showing up- and down-regulation of important proteins in penile tissues by cavernous nerve resection (CNR) as compared with sham operation (a: p<0.05).

    Fig. 3
    Immunohistochemical staining of annexin A4 in cavernous sections obtained from the penile tissue 2 weeks after sham or cavernous nerve resection (CNR) operation ((A) sham ×200, (B) CNR ×200, arrow: endothelial cells, arrow head: smooth muscle cells).

    Tables

    Table 1
    Up- and down-regulated proteins in cavernous tissue after cavernous nerve resection

    Notes

    This work was supported by the Korea Research Foundation Grant funded by the Korean Government (MOEHRD, Basic Research Promotion Fund) (KRF-2007-521-E00089).

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    MeSH Terms
    Animal Experimentation
    Animals
    Annexin A4
    Apoptosis
    Caves
    Digestion
    Erectile Dysfunction
    Fibrosis
    Gels
    Glycolysis
    Humans
    Immunohistochemistry
    Male
    Mass Spectrometry
    Prostatectomy
    Proteins
    Proteomics
    Pyruvate Kinase
    Rats
    Rats, Inbred WKY
    Salicylamides
    Silver Nitrate
    Metrics
    Share
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