Division of Urology, Department of Organ Therapeutics, Kobe University Graduate School of Medicine, Kobe, Japan
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Abstract |
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Key words: anti-Müllerian hormone/oligozoospermia/Sertoli cell development/spermatogenesis
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Introduction |
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In the present study, we measured seminal AMH concentrations in oligozoospermic men and normal men, examining correlation between AMH and various clinical parameters.
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Materials and methods |
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Serum hormones and AMH assays
Serum samples were obtained from all patients and stored at 20°C until assays. FSH and LH were determined by chemiluminescence assays (Bayer, Wuppertal, Germany). Testosterone and estradiol were measured by radioimmunoassay (Yatoron, Tokyo, Japan). Detection limits were 0.3 mIU/ml for FSH, 0.1 mIU/ml for LH, 0.05 ng/ml for testosterone and 10 pg/ml for estradiol. Seminal plasma was obtained by centrifugation after liquefaction, frozen quickly and then stored at 80°C until assays. AMH was measured using a commercially available double-antibody enzyme-linked immunoassay (Serotec, Oxford, UK). Intra- and inter-assay coefficients of variation were 6 and 15% respectively. The lowest detectable AMH concentration was 1.8 pg/ml.
Statistical analysis was performed using the Statview statistical program for MacIntosh (Abacus Concepts Inc., Calabasas, CA, USA). Spearman's rank correlation test was used to assess correlation and the non-parametric MannWhitney test was used to assess differences between groups. P < 0.05 was regarded as indicating a significant difference.
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Results |
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Discussion |
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In humans, large amounts of AMH are produced during fetal and post-natal testicular development (Tran et al., 1987; Voutilainen and Miller, 1987
; Kuroda et al., 1990
; Sweeney et al., 1997
). AMH expression is sharply down-regulated at the time when primary spermatocytes appear. This decrease in AMH expression reflects terminal differentiation of Sertoli cells and is probably only partially dependent upon a regulatory factor associated with the onset of meiosis (Meyts et al., 1999
). While some AMH is secreted after puberty, in adults the concentrations of AMH are significantly higher in seminal plasma than in serum (Fenichel et al., 1999
). This suggests that AMH is preferentially secreted at the apical pole of the Sertoli cell, toward the seminiferous tubular lumen (Fallat et al., 1996
; Fenichel et al., 1999
). Maddocks and Sharpe also reported a progressive reduction with age in the secretion of inhibin-
via the base of Sertoli cells into the interstium and an increase via the apex of the Sertoli cells (Maddocks and Sharpe, 1990
). The mechanism of the directional change remains unknown. Baarends et al. suggested that the presence of more developmentally advanced spermatogenic cells may direct AMH secretion toward the Sertoli cell apex in relation to specific maturational stages of the seminiferous epithelium (Baarends et al., 1995
).
In the present study, we demonstrated that the seminal AMH concentration correlated with sperm concentration and that the seminal concentration of AMH in normal control subjects was significantly higher than in oligozoospermic men. Fenichel et al. also demonstrated that, in non-obstructed azoospermia, the seminal AMH concentration was lower than in fertile donors (Fenichel et al., 1999). Therefore, we believe that the seminal AMH concentration is a marker of spermatogenesis, and that seminal AMH secreted from the apical aspect of Sertoli cells may be involved in sperm production and germ cell proliferation. Although Fallat et al. demonstrated an inverse relationship between seminal AMH concentration and motility index (Fallat et al., 1996
), we did not find that relationship.
While a low AMH concentration in seminal plasma appears to be related to spermatogenic dysfunction, a low concentration also may suggest immaturity of Sertoli cells. A Sertoli cell with immature characteristics cannot secrete AMH through its apical layer into seminiferous tubules; instead, any secretion proceeds through the basal layer into the interstium. As a consequence, seminal concentrations in mature Sertoli cells are higher than in serum, and than those in seminal plasma derived from immature Sertoli cells. The seminal concentration of AMH could serve as a marker of Sertoli cell maturity. In addition, severe spermatogenic impairment in human seminiferous tubules is associated with a population of Sertoli cells that exhibit a prepubertal stage of development (Steger et al., 1996). The seminal concentration of AMH should be helpful for determining the extent to which Sertoli cell immaturity is associated with defective spermatogenesis or contributes to spermatogenic dysfunction.
In conclusion, seminal concentration of AMH correlates with sperm production and may be a good marker for spermatogenesis, as well as Sertoli cell development.
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Notes |
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References |
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Submitted on September 7, 2001; accepted on November 21, 2001.