Hart N H, Cook M
J Exp Zool. 1977 Jan;199(1):109-27. doi: 10.1002/jez.1401990113.
The ontogeny of esterase isozymes in Brachydanio rerio (zebra danio), Brachydanio albolineatus (pearl danio), and hybrids formed by their reciprocal crosses was investigated using polyacrylamide disc electrophoresis. Seven esterase isozymes were identified in each species from the unfertilized egg stage to nine days posthatch. Electrophoretic analysis of qualitative changes in enzyme pattern indicated that some esterases were present at all stages of development while other esterases abruptly appeared at a specific stage of morphological differentiation. The esterases of both species were classified on the basis differential substrate and inhibitor specificities. In developing hybrids formed by B. rerio eggs inseminated with B. albolineatus sperm, the maternal isozyme pattern persisted until Stage 17 (gastrulation). Embryonic extracts from Stage 17 onward showed a slow-moving, DFP-sensitive carboxylesterase of paternal origin. In developing hybrids formed by B. albolineatus eggs inseminated with B. rerio sperm, a paternal contribution to the esterase pattern was probably present by the end of gastrulation; esterase activity of distinctively paternal origin was present by Stage 22 (retinal pigmentation) The maternal contribution to the total esterase profile appeared to remain high through hatching. Additional evidence for gene activity at gastrulation was obtained in experiments utilizing actinomycin-D and cycloheximide. Results of exposing embryos of B. rerio to 15 mug/ml of actinomycin-D indicated that transcription of the template RNA coding for cholinesterase occurred during gastrulation or some 20-30 hours prior to the appearance of the isozyme at Stage 22. This template RNA was translated sometime during that 10-hour interval immediately preceding Stage 22.
利用聚丙烯酰胺圆盘电泳法,对斑马鱼、珍珠斑马及其正反交杂种酯酶同工酶的个体发育进行了研究。在每个物种中,从未受精卵阶段到孵化后九天共鉴定出七种酯酶同工酶。对酶谱定性变化的电泳分析表明,一些酯酶在发育的各个阶段均有出现,而其他酯酶则在形态分化的特定阶段突然出现。根据底物和抑制剂特异性的差异,对两个物种的酯酶进行了分类。在用珍珠斑马精子授精的斑马鱼卵发育形成的杂种中,母本同工酶谱一直持续到17期(原肠胚形成期)。从17期开始的胚胎提取物显示出一种来自父本的、迁移缓慢且对二异丙基氟磷酸敏感的羧酸酯酶。在用斑马鱼精子授精的珍珠斑马卵发育形成的杂种中,到原肠胚形成期末可能已经出现了父本对酯酶谱的贡献;到22期(视网膜色素沉着期)出现了明显来自父本的酯酶活性。在孵化前,母本对总酯酶谱的贡献似乎一直很高。在使用放线菌素-D和环己酰亚胺的实验中,获得了原肠胚形成期基因活性的额外证据。将斑马鱼胚胎暴露于15微克/毫升放线菌素-D的结果表明,编码胆碱酯酶的模板RNA的转录发生在原肠胚形成期,即22期同工酶出现前约20 - 30小时。这种模板RNA在紧接22期之前的10小时内的某个时间被翻译。