CYTOGENETIC ANALYSIS OF SPERM USING THE HUMSTER SYSTEM

这些研究的长期目标是调查遗传因素对 以及环境因素对人类精子的产生和功能的影响。 导致精子发生停滞和减数分裂异常的突变导致 不育或生育力低下。 体质染色体异常，如 由于易位在胎儿损耗中具有重要的病因学意义， 出生和发育缺陷。 环境诱变剂和致畸剂 人们在工业或治疗环境中可能接触到的化学物质， 可能对生育力和出生缺陷的起源有类似的影响。 最近，已经可以评估精子的受精能力， 在体外系统中进行精子染色体的核型分析。 卵收集自超数排卵的金黄仓鼠（Mesocricetus auratus）并通过轻微的胰蛋白酶消化去除透明带。 当 将这种无透明带仓鼠卵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 原核可以进展到中期（因此，杂种术语， Humster系统）。 精子头部不致密的卵子比例 精子穿透试验（SPA）是精子穿透试验（SPA）的一种。 后者通常被认为是渗透性的指标 精子样本的样本 秋水仙胺处理穿透卵 促进原核浓缩成中期板， 在应用适当的显带程序后进行核型分析（例如， Q显带）。 我们建议使用Humster系统来研究以下几组男性 有不育和染色体异常精子产生的风险， 遗传和环境原因：（1）正常体质的男性 核型不育的男性，（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）霍奇金病男性 谁是进入纪念医院的化疗方案，（5）男性与 睾丸生殖细胞肿瘤的患者 放射治疗方案，以及（6）意外或 职业性暴露于致染色体断裂剂、诱变剂或致畸剂。 结果 这些研究有望更好地定义 精子以及获得对不孕症和生育的病因学的见解 缺陷 SPA的评价技术以及精子分析 染色体在我们的实验室中发育良好。 我们的临床资源 （纪念医院和纽约医院）将确保提供 适合本研究的患者人群。

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.