Adler I D, Bishop J, Lowe X, Schmid T E, Schriever-Schwemmer G, Xu W, Wyrobek A J
Institut für Säugetiergenetik, GSF Forschungszentrum für Umwelt und Gesundheit GmbH, Oberschleissheim, Germany.
Mutat Res. 1996 Dec;372(2):259-68. doi: 10.1016/s0027-5107(96)00145-5.
This study was designed to evaluate the frequency of aneuploid sperm in young adult mice of the genotype (102/E1 x C3H/E1)F1 determined by the fluorescence in situ hybridization (FISH) procedure and to evaluate the frequencies of aneuploid sperm observed by FISH compared with the frequencies of aneuploid offspring. Three-chromosome FISH was applied to determine the fractions of hyperhaploid and diploid sperm with DNA probes specific for chromosomes X, Y and 8. The animals were treated with three common solvents. Sperm smears were prepared for FISH by two similar protocols and were scored by different persons and in two different laboratories. There were no significant differences between scorers or laboratories. The frequencies of the sex chromosome aneuploidies in sperm (Y-Y and X-Y) were compared to the frequencies of mice carrying sex chromosome aneuploidy among controls of the heritable translocation assay in studies conducted from 1975-1995. To identify aneuploid individuals, untreated males and females of the genotype (102/E1 x C3H/E1)F1 were mated to assess their fertility by observing three consecutive litters. Semisterile and sterile animals were further analysed by meiotic cytogenetics and by karyotyping to determine the incidence of reciprocal translocations and sex chromosome aneuploidies (XXY and XYY). Based on the analysis of 175247 sperm and 9840 progeny, the frequency of Y-Y sperm was 0.01% while 0.03% of the offspring were XYY. The frequency of X-Y sperm was 0.005% while 0.02% of the offspring were XXY. The frequencies of aneuploid sex chromosomes were not significantly different between sperm and offspring. This allows two conclusions. First, there was no detectable prenatal selection against these sex-chromosomal aneuploid offspring, and second, germ cell aneuploidy can be reliably determined in mice by sperm FISH analyses.
本研究旨在通过荧光原位杂交（FISH）程序评估基因型为(102/E1 x C3H/E1)F1的年轻成年小鼠中非整倍体精子的频率，并将FISH观察到的非整倍体精子频率与非整倍体后代的频率进行比较。应用三染色体FISH，使用针对X、Y和8号染色体的DNA探针来确定超单倍体和二倍体精子的比例。对动物使用三种常见溶剂进行处理。通过两种相似的方案制备用于FISH的精子涂片，并由不同人员在两个不同实验室进行评分。评分者或实验室之间没有显著差异。将精子中性染色体非整倍体（Y-Y和X-Y）的频率与1975 - 1995年进行的遗传易位试验对照组中携带性染色体非整倍体的小鼠频率进行比较。为了鉴定非整倍体个体，将基因型为(102/E1 x C3H/E1)F1的未处理雄性和雌性进行交配，通过观察连续三窝仔鼠来评估它们的生育能力。对半不育和不育动物进一步进行减数分裂细胞遗传学分析和核型分析，以确定相互易位和性染色体非整倍体（XXY和XYY）的发生率。基于对175247个精子和9840个后代的分析，Y-Y精子的频率为0.01%，而0.03%的后代为XYY。X-Y精子的频率为0.005%，而0.02%的后代为XXY。精子和后代中性染色体非整倍体的频率没有显著差异。由此可得出两个结论。第一，未检测到针对这些性染色体非整倍体后代的产前选择；第二，通过精子FISH分析可以可靠地确定小鼠生殖细胞的非整倍体情况。
