Rahman Sk Mustafizur, Majhi Sullip Kumar, Suzuki Toru, Matsukawa Shingo, Strüssmann Carlos Augusto, Takai Rikuo
Department of Food Science and Technology, Tokyo University of Marine Science and Technology, Konan 4-5-7, Minato-Ku, Tokyo 108-8477, Japan.
Cryobiology. 2008 Oct;57(2):170-4. doi: 10.1016/j.cryobiol.2008.08.002. Epub 2008 Aug 12.
The purpose of this study was to examine the suitability of cryoprotectant agent (CPA) impregnation protocols for the embryos of Japanese whiting (Sillago japonica), a small-sized, easy-to-rear, and prolific marine fish which may constitute a suitable experimental material for the development of cryopreservation methods for fish embryos. Our immediate goals were to assess the toxicity and permeability of various CPAs to whiting embryos of different developmental stages. Exposure of gastrula, somites, tail elongation, and pre-hatching embryos to 10%, 15%, and 20% solutions of propylene glycol (PG), methanol (MeOH), dimethyl sulfoxide (Me2SO), dimethylformamide (DFA), ethylene glycol (EG), and glycerol (Gly) in artificial sea water (ASW; 33 psu) for 20 min revealed that CPA toxicity for whiting embryos increased in the order of PG<Me2SO<DFA<EG<MeOH<Gly. Relative CPA permeability, estimated by proton nuclear magnetic resonance spectroscopy, showed the same trend as toxicity except for Gly, which was highly toxic but showed only moderate permeability. There were no marked differences in CPA tolerance between developmental stages except for a slight decrease in pre-hatching embryos. Stepwise (20% x 5 steps) impregnation with CPA mixtures of 20-25% PG with 10-15% DFA, Me2SO, MeOH, or EG in ASW were well tolerated by the embryos. Overall, the results of toxicity and permeability suggest that PG, MeOH, and EG could be useful for the development of CPA solutions for whiting embryos.
本研究的目的是检验低温保护剂（CPA）浸渍方案对日本银鲈（Sillago japonica）胚胎的适用性。日本银鲈是一种小型、易于养殖且繁殖力强的海鱼，可能是开发鱼类胚胎冷冻保存方法的合适实验材料。我们的近期目标是评估各种CPA对不同发育阶段银鲈胚胎的毒性和渗透性。将原肠胚、体节期、尾芽期和孵化前胚胎暴露于人工海水（ASW；33 psu）中10%、15%和20%的丙二醇（PG）、甲醇（MeOH）、二甲基亚砜（Me2SO）、二甲基甲酰胺（DFA）、乙二醇（EG）和甘油（Gly）溶液中20分钟，结果显示，银鲈胚胎对CPA的毒性按PG＜Me2SO＜DFA＜EG＜MeOH＜Gly的顺序增加。通过质子核磁共振光谱法估计的相对CPA渗透性显示出与毒性相同的趋势，但Gly除外，Gly毒性高但渗透性仅为中等。除孵化前胚胎略有下降外，不同发育阶段对CPA的耐受性没有明显差异。胚胎对在ASW中用20 - 25% PG与10 - 15% DFA、Me2SO、MeOH或EG的CPA混合物进行逐步（20%×5步）浸渍耐受性良好。总体而言，毒性和渗透性结果表明，PG、MeOH和EG可用于开发银鲈胚胎的CPA溶液。
