Mailhes J B, Preston R J, Yuan Z P, Payne H S
Department of Obstetrics and Gynecology, Louisiana State University Medical Center, Shreveport 71130.
Mutat Res. 1988 Mar;198(1):145-52. doi: 10.1016/0027-5107(88)90049-8.
Our initial objective was to develop an in vivo mammalian, female aneuploid assay that is consistent, time efficient, and that yields a large number of oocytes amenable to objective analyses. Subsequently, we desired to use such an assay for identifying chemicals and dosages that could increase the incidence of aneuploidy in mouse metaphase II oocytes. The experimental protocol involved superovulating CD-1 mice with PMS; HCG was given 48 h later. At the time of HCG injection, different dosages od diethylstilbestrol diphosphate, cadmium chloride, chloral hydrate, or colchicine were injected intraperitoneally. 17 h later, oocytes were collected and fixed prior to C-banding the chromosomes. The procedure required about 3 h to process oocytes from 25 mice and yielded over 100 analyzable metaphase II oocytes. Colchicine was the only compound tested that resulted in a statistically significant (P less than 0.01) increase in hyperploid (N greater than 20) oocytes over controls. The incidence of hyperploid oocytes in the colchicine group was 2/167, 1/182, 21/220, and 38/202, for control, 0.1 mg/kg, 0.2 mg/kg, and 0.3 mg/kg, respectively. This assay appears sensitive for aneuploidy detection but requires further validation.
我们最初的目标是开发一种体内哺乳动物雌性非整倍体检测方法，该方法具有一致性、省时性，并且能产生大量适合客观分析的卵母细胞。随后，我们希望使用这样一种检测方法来鉴定能够增加小鼠中期II卵母细胞非整倍体发生率的化学物质和剂量。实验方案包括用孕马血清促性腺激素（PMS）对CD-1小鼠进行超排卵；48小时后注射人绒毛膜促性腺激素（HCG）。在注射HCG时，将不同剂量的二乙基己烯雌酚二磷酸酯、氯化镉、水合氯醛或秋水仙碱腹腔注射。17小时后，收集卵母细胞并固定，然后对染色体进行C显带。该程序处理25只小鼠的卵母细胞大约需要3小时，并产生100多个可分析的中期II卵母细胞。秋水仙碱是唯一测试的化合物，与对照组相比，其超倍体（N大于20）卵母细胞数量有统计学意义的显著增加（P小于0.01）。秋水仙碱组中超倍体卵母细胞的发生率分别为：对照组为2/167，0.1mg/kg组为1/182，0.2mg/kg组为21/220，0.3mg/kg组为38/202。该检测方法似乎对非整倍体检测敏感，但需要进一步验证。
