Mamsen L S, Petersen T S, Jeppesen J V, Møllgård K, Grøndahl M L, Larsen A, Ernst E, Oxvig C, Kumar A, Kalra B, Andersen C Y
Laboratory of Reproductive Biology, Section 5712, The Juliane Marie Centre for Women, Children and Reproduction, University Hospital of Copenhagen, University of Copenhagen, Rigshospitalet, Copenhagen DK-2100, Denmark
Laboratory of Reproductive Biology, Section 5712, The Juliane Marie Centre for Women, Children and Reproduction, University Hospital of Copenhagen, University of Copenhagen, Rigshospitalet, Copenhagen DK-2100, Denmark.
Mol Hum Reprod. 2015 Jul;21(7):571-82. doi: 10.1093/molehr/gav024. Epub 2015 Apr 28.
From early embryonic life, anti-Müllerian hormone (AMH) is produced by Sertoli cells and is essential for male sex differentiation. In females, AMH is produced by immature granulosa cells (GCs) but a definitive function in females is uncertain. We have assessed the cellular localization and specificity of a panel of five novel high-affinity AMH monoclonal antibodies. Two recognize the mature C-terminal form of AMH, whereas three recognize the active pro-mature form of AMH in human tissue. The antibodies were tested on fetal male testicular and mesonephric tissue aged 8-19 weeks post conception (pc), fetal male serum aged 16-26 weeks pc and human immature GCs by immunofluorescence, immunohistochemistry, ELISA and western blotting. The active pro-mature forms of AMH were expressed in both Sertoli cells from human fetal testis and human immature GCs. In contrast, the mature C-terminal form of AMH was hardly detected in Sertoli cells, but was readily detected in GCs. This particular form was also located to the nucleus in GCs, whereas the other investigated AMH forms remained in the cytoplasm. Interestingly, the distribution of the AMH forms in the fetal serum of boys showed that the fraction of inactive precursor AMH only accounted for 4.5% ± 0.6 (mean ± SD) of the total AMH measured, and the remaining AMH was the active pro-mature form. Furthermore, western blot analysis demonstrated a number of previously unrecognized molecular forms of AMH. The present findings suggest that processing of AMH is a tightly regulated process, which is likely to be important for the function of AMH and which differs between the two sexes.
从胚胎早期开始，抗苗勒管激素（AMH）由支持细胞产生，对男性性别分化至关重要。在女性中，AMH由未成熟的颗粒细胞（GCs）产生，但其在女性中的明确功能尚不确定。我们评估了一组五种新型高亲和力AMH单克隆抗体的细胞定位和特异性。两种抗体识别AMH的成熟C末端形式，而三种抗体识别人类组织中AMH的活性前体成熟形式。通过免疫荧光、免疫组织化学、ELISA和蛋白质印迹法，对妊娠8 - 19周（pc）的胎儿男性睾丸和中肾组织、妊娠16 - 26周pc的胎儿男性血清以及人类未成熟GCs进行了抗体检测。AMH的活性前体成熟形式在人类胎儿睾丸的支持细胞和人类未成熟GCs中均有表达。相比之下，AMH的成熟C末端形式在支持细胞中几乎未检测到，但在GCs中很容易检测到。这种特定形式在GCs中也定位于细胞核，而其他研究的AMH形式则保留在细胞质中。有趣的是，男孩胎儿血清中AMH形式的分布表明，无活性前体AMH的比例仅占所测总AMH的4.5%±0.6（平均值±标准差），其余的AMH是活性前体成熟形式。此外，蛋白质印迹分析显示了一些以前未识别的AMH分子形式。目前的研究结果表明，AMH的加工是一个严格调控的过程，这可能对AMH的功能很重要，并且在两性之间存在差异。
