Dos Santos Matheus Pereira, do Nascimento Nivaldo Ferreira, Yasui George Shigueki, Pereira Nycolas Levy, Fujimoto Takafumi, Senhorini José Augusto, Nakaghi Laura Satiko Okada
Aquaculture Center,São Paulo State University,Jaboticabal,Brazil.
National Center for Research and Conservation of Continental Fish,Chico Mendes Institute of Biodiversity Conservation,Pirassununga,Brazil.
Zygote. 2018 Feb;26(1):89-98. doi: 10.1017/S0967199417000739. Epub 2018 Jan 15.
In fish, many factors can affect reproduction during in vitro fertilization, therefore determination of the factors that affect affecting gamete quality is needed. However, few studies have focused on gamete quality and the ploidy status. This study was conducted to elucidate whether oocyte storage can affect ploidy status, survival, and embryo viability in the characid species Astyanax altiparanae. Oocytes were stored in Dulbecco's phosphate-buffered saline (PBS) at 26°C, then aliquots were fertilized immediately after extrusion (control) and also after 60, 120, 180, and 240 min of storage. Fertilization and hatching rates were measured, and the developmental stages were analyzed at each stage before describing the main abnormalities. Ploidy status was analyzed by flow cytometry and blood smear. In the control group, 100% of the samples were diploid. After treatment for 60 min, 95.56 ± 4.44% samples were diploid and 4.44 ± 4.44% were triploid. After 120 min, 94.44 ± 9.62% of the samples was diploid and 5.56 ± 5.56% were triploid; 100% of the samples were diploid after 180 min and, after 240 min, there was no survival. In other treatments, the highest percentage of hatching was after 60 min (88.93 ± 5.15%; P = 0.015), and treatment with 180 min storage resulted in the highest percentage of abnormal larvae (95.76 ± 12.67%; P = 0.012). These results show that oocyte storage can affect ploidy status and may be an interesting parameter for analysis in studies on chromosome set manipulation and micromanipulation.
在鱼类中，许多因素会影响体外受精过程中的繁殖，因此需要确定影响配子质量的因素。然而，很少有研究关注配子质量和倍性状态。本研究旨在阐明卵母细胞储存是否会影响脂鲤科物种高身丽脂鲤的倍性状态、存活率和胚胎活力。将卵母细胞储存在26℃的杜氏磷酸盐缓冲盐水（PBS）中，然后在卵母细胞挤出后立即进行受精（对照组），以及在储存60、120、180和240分钟后进行受精。测量受精率和孵化率，并在描述主要异常情况之前分析每个阶段的发育阶段。通过流式细胞术和血涂片分析倍性状态。在对照组中，100%的样本为二倍体。处理60分钟后，95.56±4.44%的样本为二倍体，4.44±4.44%为三倍体。120分钟后，94.44±9.62%的样本为二倍体，5.56±5.56%为三倍体；180分钟后100%的样本为二倍体，240分钟后则无存活个体。在其他处理中，孵化率最高的是60分钟后（88.93±5.15%；P=0.015），储存180分钟的处理导致异常幼虫的比例最高（95.76±12.67%；P=0.012）。这些结果表明，卵母细胞储存会影响倍性状态，可能是染色体组操作和显微操作研究中一个值得分析的有趣参数。
