CYTOGENETIC ANALYSIS OF SPERM USING THE HUMSTER SYSTEM

这些研究的长期目标是调查遗传基因的影响 以及环境因素对人类精子的产生和功能的影响。 导致生精停止和减数分裂异常的突变会导致 不孕不育或不育。体质染色体异常，如 由于易位在胎儿损耗中具有病因学意义，仍然 出生和发育缺陷。环境致突变物和致畸剂 人们也可能在工业或治疗环境中接触到这些 会对生育能力和出生缺陷的来源产生类似的影响。 最近，评估精子的受精性和 在体外系统中对精子染色体进行核型分析。 卵是从超数排卵的金黄仓鼠(中仓鼠)采集的 金鱼)和通过轻微胰酶消化去除透明带。什么时候 这种无透明带的仓鼠卵子与体外获能的人混合 精子，精子穿透卵子。卵子和精子核发育成 可以进展到中期的原核(因此，混合术语， Humster系统)。解冻精子头的卵子比例 (以百分比表示)被称为精子穿透试验(SPA)。 后者通常被认为是渗透性的指标 给出的精子样本。Colcemid-穿透性卵子的治疗 促进原核凝聚成中期板，这可能是 在应用适当的显带程序后进行核型分析(例如 Q-带)。 我们建议使用Humster系统研究以下几组人 有不孕不育和产生染色体异常精子的风险 遗传和环境原因：(1)体质正常的男性 不育的核型；(2)罕见突变纯合子的男性 导致减数分裂紊乱并导致无精子症或少精子症， (3)携带染色体重排的男性；(4)霍奇金氏病患者 参加纪念医院化疗方案的人，(5)患有 纪念医院收治的睾丸生殖细胞肿瘤 放射治疗方案，以及(6)意外或 职业接触致裂剂、诱变剂或致畸剂的。结果： 这些研究有望更好地定义先天性心脏病的遗传学 精子以及对不孕不育和生育的病因的洞察 缺陷。SPA的评估技术以及精子的分析 我们实验室的染色体很发达。我们的临床资源 (纪念医院和纽约医院)将确保提供 适合这项研究的患者群体。

The long-term goal of these studies is to investigate the effect of genetic and environmental factors on the production and function of human sperm. Mutations that cause spermatogenic arrest and meiotic abnormalities lead to infertility or subfertility. Constitutional chromosome abnormalities such as translocations are of etiologic significance in fetal wastage, still births, and developmental defects. Environmental mutagens and teratogens to which people may be exposed in an industrial or therapeutic context also can have similar effects on fertility and origin of birth defects. Recently, it has become possible to evaluate fertilizability of sperm and to perform karyotypic analysis of sperm chromosomes in an in vitro system. Eggs are collected from superovulated golden hamsters (Mesocricetus auratus) and the zona pellucida removed by slight trypsinization. When such zona-free hamster eggs are mixed with in vitro-capacitated human sperm, the sperm penetrate the eggs. The egg and sperm nuclei develop into pronuclei which can progress to metaphase (hence, the hybrid term, the humster system). The proportion of eggs with decondensed sperm heads (expressed as percentage) is designated the sperm penetration assay (SPA). The latter is generally considered to be an indicator of the penetrability of the given sperm specimen. Colcemid-treatment of penetrated eggs promotes condensation of pronuclei into metaphase plates which can be karyotyped after application of an appropriate banding procedure (e.g. Q-banding). We propose to study, using the humster system, the following groups of men at risk for infertility and generation of chromosomally abnormal sperm due to genetic and environmental causes: (1) men with normal constitutional karyotypes who are infertile, (2) men who are homozygous for rare mutations that cause meiotic disturbances and lead to azoospermia or oligospermia, (3) men who carry chromosome rearrangements, (4) men with Hodgkin's disease who are entered on Memorial Hospital's chemotherapy protocols, (5) men with testicular germ cell tumors who are entered on Memorial Hospital's radiation therapy protocols, and (6) men who have been accidentally or occupationally exposed to clastogens, mutagens, or teratogens. Results of these studies are expected to permit better definition of the genetics of sperm as well as gain insights into the etiology of infertility and birth defects. Techniques for evaluation of SPA as well as analysis of sperm chromosomes are well developed in our laboratory. Our clinical resources (Memorial Hospital and New York Hospital) will ensure the availability of appropriate patient populations for this study.