体外受精时，冷冻解冻的未分选和按性别分选的公羊精子的配子共孵育时间对体外成熟的成年和青春期前母羊卵母细胞发育的影响。
The effect of gamete co-incubation time during in vitro fertilization with frozen-thawed unsorted and sex-sorted ram spermatozoa on the development of in vitro matured adult and prepubertal ewe oocytes.
作者信息
Morton Katherine M, Catt Sally L, Hollinshead Fiona K, Maxwell W M Chis, Evans Gareth
机构信息
Centre for Advanced Technologies in Animal Genetics and Reproduction (ReproGen), Faculty of Veterinary Science, The University of Sydney, NSW 2006, Australia.
出版信息
Theriogenology. 2005 Jul 15;64(2):363-77. doi: 10.1016/j.theriogenology.2004.12.004.
In vitro matured adult (Experiment 1) and prepubertal (Experiment 2) ewe oocytes were co-incubated with unsorted or sex-sorted frozen-thawed spermatozoa for 2 to 3 h (short) or 18 to 20 h (long) to determine the effects of reducing the gamete co-incubation time during IVF on subsequent embryonic development in vitro. For oocytes derived from adult ewes, there were no differences in oocyte fertilization and cleavage at 24 h post insemination (hpi) between types of spermatozoa or co-incubation times (P > 0.05). By 48 hpi, oocyte cleavage was higher after a short (390/602, 64.8%) compared with a long (381/617, 61.7%) co-incubation (P < 0.05), and was not significantly different for unsorted (266/372, 71.5%) and sex-sorted (505/849, 59.9%) spermatozoa. Blastocyst formation from cleaved oocytes was similar for unsorted (150/266, 56.4%) and sex-sorted (295/505, 58.4%) spermatozoa, but was higher after a short (240/390, 61.5%) than long (205/381, 53.8%) co-incubation (P < 0.05). Oocyte development to the blastocyst stage was not different for unsorted (150/372; 40.3%) and sex-sorted (295/847; 34.8%) spermatozoa but was significantly increased by a short (240/602, 39.9%) compared with a long (205/617, 33.2%) co-incubation. Fertilization of oocytes from prepubertal ewes was similar for types of spermatozoa and for duration of co-incubation. Oocyte cleavage (48 hpi) was similar for a short (241/377, 63.9%) and long (226/349, 64.8%) co-incubation with unsorted spermatozoa, but was increased (P < 0.05) by a long co-incubation (286/500, 57.2% versus 163/517, 31.5%) with sex-sorted spermatozoa. Blastocyst formation from cleaved oocytes was similar for unsorted (230/467, 49.3%) and sex-sorted (186/449, 41.4%) spermatozoa, and a short (200/404, 49.5%) or long (216/512, 42.1%) co-incubation. However, oocyte development to the blastocyst stage was higher (P < 0.05) after IVF with unsorted (230/726, 37.1%) than sex-sorted (186/1017, 18.3%) spermatozoa. Reducing the duration of gamete co-incubation did not deleteriously affect the in vitro development of adult and prepubertal ewe derived oocytes after IVF with unsorted and sex-sorted spermatozoa. In general, sex-sorting had no substantial influence on fertilization and embryo development rates.
将体外成熟的成年母羊（实验1）和青春期前母羊（实验2）的卵母细胞与未分选或经性别分选的冻融精子共同孵育2至3小时（短时间）或18至20小时（长时间），以确定在体外受精过程中缩短配子共同孵育时间对随后体外胚胎发育的影响。对于成年母羊来源的卵母细胞，授精后24小时（hpi），不同类型精子或不同共同孵育时间的卵母细胞受精率和卵裂率无差异（P>0.05）。到48 hpi时，短时间（390/602，64.8%）共同孵育后的卵母细胞卵裂率高于长时间（381/617，61.7%）共同孵育后的（P<0.05），未分选精子（266/372，71.5%）和经性别分选精子（505/849，59.9%）的卵裂率无显著差异。未分选精子（150/266，56.4%）和经性别分选精子（295/505，58.4%）形成囊胚的卵裂卵相似，但短时间（240/390，61.5%）共同孵育后的囊胚形成率高于长时间（205/381，53.8%）共同孵育后的（P<0.05）。未分选精子（150/372；40.3%）和经性别分选精子（295/847；34.8%）使卵母细胞发育到囊胚阶段无差异，但短时间（240/602，39.9%）共同孵育后的显著高于长时间（205/617，33.2%）共同孵育后的。青春期前母羊卵母细胞的受精情况在不同类型精子和不同共同孵育时间方面相似。未分选精子短时间（241/377，63.9%）和长时间（226/349，64.8%）共同孵育后的卵母细胞卵裂率相似，但经性别分选精子长时间（286/500，57.2%对163/517，31.5%）共同孵育后的卵裂率增加（P<0.05）。未分选精子（230/467，49.3%）和经性别分选精子（186/449，41.4%）以及短时间（200/404，49.5%）或长时间（216/512，42.1%）共同孵育后的卵裂卵形成囊胚的情况相似。然而，未分选精子（230/726,37.1%）体外受精后卵母细胞发育到囊胚阶段的比例高于经性别分选精子（186/1017，18.3%）（P<0.05）。缩短配子共同孵育时间对成年和青春期前母羊来源的卵母细胞在与未分选和经性别分选精子体外受精后的体外发育无有害影响。总体而言，性别分选对受精率和胚胎发育率无实质性影响。
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