Genetic imprinting during impaired spermatogenesis
Genetic imprinting during impaired spermatogenesis
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DOI：
10.1093/molehr/gal040
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发表时间：
2006-06-01
影响因子：
4
通讯作者：
Steger, Klaus
中科院分区：
文献类型：
--
作者：
Hartmann, Sonja;Bergmann, Martin;Steger, Klaus
遗传印迹障碍被认为是辅助生殖技术（ART）中潜在的遗传风险，大多数自然选择机制被绕过。由于目前只有有限的信息，基因组印迹在破坏性精子发生，我们分析了印迹状态的父系甲基化基因H19在各种生殖细胞群来自曲细精管表现出受损的精子发生。采用激光显微切割技术从人睾丸石蜡切片中分离出不同类型的生殖细胞。虽然通过甲基化特异性PCR（M-PCR）研究了母源印迹基因SNRPN的甲基化状态以建立分离方法，但通过基于单链构象的方法分析了H19的甲基化。由于支持细胞特异性波形蛋白免疫组织化学染色在生殖细胞激光显微切割前被排除。我们证明了H19的正确遗传印记，即使在精原细胞水平的生精小管中选择的精原细胞中也表现出精子发生停滞，没有证据表明用于ICSI的不育男性精子中存在不正确的基因组印记。
Disorders in genetic imprinting are discussed as potential genetic risk in assisted reproduction technology (ART), where most of the natural selection mechanisms are bypassed. As currently only limited information about genomic imprinting in disruptive spermatogenesis is available, we analysed the imprinting state of the paternally methylated gene H19 in various germ cell populations derived from seminiferous tubules exhibiting impaired spermatogenesis. Different germ cell types were isolated by laser microdissection from human testicular paraffin sections. Although the methylation state of the maternally imprinted gene SNRPN was investigated by methylation-specific PCR (M-PCR) to establish the isolation method, methylation of H19 was analysed by a single-strand conformation-based method. Contamination by somatic Sertoli cells was excluded because of Sertoli cell-specific vimentin immunohistochemistry before germ cell laser microdissection. We demonstrate correct genetic imprints for H19 even in spermatogonia selected from seminiferous tubules exhibiting spermatogenic arrest at the level of spermatogonia, providing no evidence for incorrect genomic imprinting in spermatozoa from infertile men used for ICSI.