水产学报2012，Vol.36Issue(11)：1717-1724,8.DOI:10.3724/SP.J.1231.2012.28078
超低温冷冻对中华绒螯蟹胚胎形态结构的影响
Effects of cryopreservation on exterior shape and interior structure of Chinese mitten crab (Eriocheir sinensis) embryos
摘要
Abstract
In order to deeply understand the effects of cryopreservation on the configuration of embryos, we chose the Eriocheir sinensis as the object, and observed the exterior shapes and interior structure of different developmental stages embryos after treatment with cryoprotectants and cryopreservation by means of microscope, slice and electron microscopy. Results showed that: (1) Cleavage embryos swelled clearly after treatment with vitrifying solutions by two-step methods, exterior shape of the embryos had no significant difference compared to the control group by three-step equilibration methods, some yolk overflowed from the inner cells and cells were damaged seriously after cryopreservation. (2) Exterior shape of gastrula stage embryos was similar to the control group after treatment with two-step equilibration methods, surface of gastrula stage embryo treated with cryoprotectants crinkled and shaped reticulation structure by scanning electron microscopy (SEM), white gobbet appeared in cells of embryos and fringe of cell was coarse and protuberant by transmission electron microscopy (TEM). After cryopreservation, color of embryos was pink, cell became opaque, the brim of cell membrane was blurry and shaped floss, segmental yolk was disintegrated into granule, the surface of embryos was sunken and wrinkled. (3) Cell membrane of embryos broke off, ice holes of different size appeared inside embryos, visible vacuoles appeared inside cells by slice observation. Ice holes in cells were very clear and some vacuoles appeared, about 80% mitochondria dissolved, breach of cells was very obvious by TEM. Exterior shape of the original zoea stage embryos had no difference compared to that of the control group after treatment with cryoprotectants by microscope and SEM. After cryopreservation, above 95% tissues shaped dispersion, partial yolk was disintegrated into granule, cell membrane of embryos broke off, ice holes appeared inside embryos. Surface of 90% embryos was sunken and wrinkled, but the cuticle of 10% embryos retained slick and intact by SEM. The results implied that the original zoea stage of embryos was the appropriate stage for cryopreservation.关键词
中华绒螯蟹/胚胎/形态结构/超低温冷冻Key words
Eriocheir sinensis/ cryopreservation/ embryo/ configuration分类
农业科技引用本文复制引用
黄晓荣,庄平,章龙珍,冯广朋,刘鉴毅,张涛,赵峰,侯俊利..超低温冷冻对中华绒螯蟹胚胎形态结构的影响[J].水产学报,2012,36(11):1717-1724,8.基金项目
中央级公益性科研院所基本科研业务费专项资金项目(2011T04) （2011T04）
公益性行业(农业)科研专项经费项目(201203065 （农业）
200903048-07) （）
