Baccetti Baccio, Collodel Giulia, Gambera Laura, Moretti Elena, Serafini Francesca, Piomboni Paola
Department of Pediatrics, Obstetrics and Reproductive Medicine, Siena University, Regional Referral Center for Male Infertility, Azienda Ospedaliera Universitaria Senese, Siena, Italy.
Fertil Steril. 2005 Jul;84(1):123-9. doi: 10.1016/j.fertnstert.2005.01.128.
To perform fluorescence in situ hybridization (FISH) and molecular analysis in patients with the genetic sperm defect "dysplasia of the fibrous sheath" (DFS).
Retrospective study.
Regional Referral Center for Male Infertility, Siena, Italy.
PATIENT(S): Twelve infertile patients with DFS sperm defects.
INTERVENTION(S): Family history, lymphocytic karyotype, physical and hormonal assays, semen analysis.
MAIN OUTCOME MEASURE(S): The DFS sperm phenotype was defined by light, fluorescent, and electron microscopy. Sperm chromosomal constitution was examined by FISH. Gene deletions were tested by polymerase chain reaction.
RESULT(S): The genetic sperm defect DFS was determined by transmission and scanning electron microscopy. Immunofluorescence staining of A-kinase anchoring protein 4 (AKAP4) showed a moderate and diffuse signal, revealing a disorganized and incompletely assembled fibrous sheath. In 11 of 12 DFS patients, polymerase chain reaction for detecting the presence of partial sequence of AKAP4/AKAP3 binding regions gave positive results. Fluorescence in situ hybridization was performed in decondensed sperm nuclei with probes for chromosomes 18, X, and Y. The mean disomy frequency of chromosome 18 was in the normal range, whereas the mean disomy frequencies of sex chromosomes and diploidies were twice those of controls.
CONCLUSION(S): These results should be considered when DFS sperm are used in assisted reproductive technology, owing to the high risk of transmission of chromosomal unbalance and of DFS sperm defects to male offspring.
对患有遗传性精子缺陷“纤维鞘发育异常”（DFS）的患者进行荧光原位杂交（FISH）和分子分析。
回顾性研究。
意大利锡耶纳男性不育区域转诊中心。
12例患有DFS精子缺陷的不育患者。
家族史、淋巴细胞核型分析、体格检查和激素测定、精液分析。
通过光学、荧光和电子显微镜确定DFS精子表型。通过FISH检查精子染色体组成。通过聚合酶链反应检测基因缺失。
通过透射和扫描电子显微镜确定遗传性精子缺陷DFS。A激酶锚定蛋白4（AKAP4）的免疫荧光染色显示中等强度的弥漫性信号，表明纤维鞘紊乱且组装不完全。在12例DFS患者中的11例中，检测AKAP4/AKAP3结合区域部分序列存在情况的聚合酶链反应结果为阳性。在去浓缩的精子核中用18号染色体、X染色体和Y染色体的探针进行荧光原位杂交。18号染色体的平均二体频率在正常范围内，而性染色体和二倍体的平均二体频率是对照组的两倍。
由于将DFS精子用于辅助生殖技术时，染色体不平衡和DFS精子缺陷传递给男性后代的风险很高，因此在使用时应考虑这些结果。
