Functional analysis using chlortetracycline fluorescence and in vitro fertilization of frozen-thawed ejaculated boar spermatozoa incubated in a protein-free chemically defined medium.
Functional analysis using chlortetracycline fluorescence and in vitro fertilization of frozen-thawed ejaculated boar spermatozoa incubated in a protein-free chemically defined medium.
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使用金霉素荧光进行功能分析，并对在无蛋白质化学成分确定的培养基中孵育的冻融射精公猪精子进行体外受精。
DOI：
10.1530/jrf.0.1040305
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发表时间：
1995
期刊：
影响因子：
--
通讯作者：
K. Niwa
中科院分区：
文献类型：
--
作者：
Wei‐Hua Wang;L. R. Abeydeera;L. Fraser;K. Niwa
卵丘封闭猪卵母细胞体外成熟，脱离卵丘细胞，在含有37.0 mmol NaHCO3 l-1和5 mmol咖啡因l-1的化学定义无蛋白培养基中与冻融的射精精子受精。当培养基中添加1 mg聚乙烷醇（PVA） ml-1时，7-12 × 10(6)个ml-1细胞比4-5 × 10(6)个ml-1细胞授精14 h后观察到穿透的卵母细胞更多，多精的发生率反映了使用的精子浓度。改变NaHCO3浓度，但将精子浓度维持在8 × 10(6)个细胞ml-1时，在含有45.83 ~ 50.25 mmol NaHCO3 l-1的培养基中，卵母细胞的穿透率明显高于37.0 ~ 41.42 mmol NaHCO3 l-1；在雄性原核形成或多精子的发生率上没有显著差异。在含有45.83 mmol NaHCO3 l-1的培养基中，0-5 mg ml-1的PVA对穿透卵母细胞的比例、雄性原核形成和多精子没有影响。然而，当评估来自三种不同公猪的精子时，与多精的发生率不同，穿透率和雄性原核形成率变化很大。在6 h时首次检测到无卵母细胞的穿透。当精子在没有卵母细胞的情况下孵育6 h时，无论培养基中是否含有PVA，精子的运动性（而不是活力）都会下降。CTC荧光分析显示，在第一个小时内，顶体完整、无能力的活细胞比例迅速下降，顶体反应、有能力的活细胞比例迅速上升。（摘要删节250字）
Cumulus-enclosed pig oocytes were matured in vitro, freed from cumulus cells, and inseminated with frozen-thawed ejaculated spermatozoa in a chemically defined protein-free medium containing 37.0 mmol NaHCO3 l-1 and 5 mmol caffeine l-1. When the medium was supplemented with 1 mg polyvinylalcohol (PVA) ml-1, more penetrated oocytes were observed 14 h after insemination with 7-12 x 10(6) cells ml-1 than with 4-5 x 10(6) cells ml-1 and the incidence of polyspermy reflected the sperm concentration used. Varying the NaHCO3 concentration but maintaining the sperm concentration at 8 x 10(6) cells ml-1 resulted in significantly more oocytes being penetrated in media containing 45.83-50.25 than 37.0-41.42 mmol NaHCO3 l-1; there were no significant differences in the incidence of either male pronuclear formation or polyspermy. In medium containing 45.83 mmol NaHCO3 l-1, the inclusion of PVA at 0-5 mg ml-1 had no effect on proportions of penetrated oocytes, male pronuclear formation or polyspermy. However, when spermatozoa from three different boars were evaluated, the penetration and male pronuclear formation rates were highly variable, unlike the incidence of polyspermy. Penetration of cumulus-free oocytes was first detected at 6 h. When spermatozoa were incubated for 6 h in the absence of oocytes, motility, but not vitality, decreased whether or not PVA was included in the medium. Chlortetracycline (CTC) fluorescence analysis of the capacitation state indicated a rapid decline in the proportion of live uncapacitated, acrosome-intact cells and a rapid rise in the proportion of live capacitated, acrosome-reacted cells during the first hour.(ABSTRACT TRUNCATED AT 250 WORDS)
影响因子：
3.6
作者：
BOATMAN, DE;ROBBINS, RS
通讯作者：
ROBBINS, RS
合成有机 pH 缓冲剂可以支持豚鼠卵受精，但不如碳酸氢盐缓冲剂有效。
DOI：
10.1002/mrd.1120190203
发表时间：
1988
期刊：
Gamete research
影响因子：
--
作者：
Bhattacharyya,A;Yanagimachi,R
通讯作者：
Yanagimachi,R