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渗透压和糖胺聚糖对牛射精精子活力、获能、顶体反应及体外受精能力的影响。

Effect of osmolality and glycosaminoglycans on motility, capacitation, acrosome reaction, and in vitro fertilizability of bovine ejaculated sperm.

作者信息

Miller D J, Hunter A G

出版信息

J Dairy Sci. 1986 Nov;69(11):2915-24. doi: 10.3168/jds.S0022-0302(86)80747-0.

DOI:10.3168/jds.S0022-0302(86)80747-0
PMID:3100592
Abstract

Bovine ejaculated semen was placed in a modified Tyrode's medium with albumin, lactate, and pyruvate. The sperm were washed three times and subjected to nine treatment in a 3 X 3 factorial arrangement. Treatments consisted of osmolality (exposure to 380 mOsmol/kg medium for 5 min, exposure to 340 or 295 mOsmol/kg medium for the entire incubation period), and the presence or absence of glycosaminoglycans (100 micrograms/ml chondroitin sulfate A or 10 micrograms/ml heparin). Sperm were examined at 4.5 h, 8 to 9 h, and 24 to 25 h of incubation (37 degrees C, 5% CO2, and 95% air). Heparin caused head-to-head agglutination of sperm, raised the percent sperm without seminal antigens over the acrosome (capacitated) by 20% at 4.5 h, and doubled the percent of acrosome-reacted sperm. However, this stimulation did not improve in vitro fertilizability. Chondroitin sulfate A tended to maintain motility, but did not affect capacitation or the acrosome reaction, possibly due to glucose inhibition. Both high osmolality treatments tended to reduce motility, especially after 24 h of incubation when the 340 osmolality treatment reduced motility by 14% over the 295 treatment. No consistent effect on capacitation was observed. The 340 and 380 osmolality treatments induced 8.6 and 6.1% more acrosome reactions by 24 h than the 295 treatment. The 340 mOsmol/kg treatment yielded insignificantly higher in vitro fertilization rates, as evidenced by development of zygotes to the two-cell stage. Lack of statistical significance was due to high variation with in vitro fertilization rates.

摘要

将牛射精后的精液置于含有白蛋白、乳酸盐和丙酮酸盐的改良Tyrode's培养基中。精子洗涤三次后,按照3×3析因设计进行九种处理。处理包括渗透压(在380毫渗摩尔/千克培养基中暴露5分钟,在整个孵育期暴露于340或295毫渗摩尔/千克培养基),以及是否存在糖胺聚糖(100微克/毫升硫酸软骨素A或10微克/毫升肝素)。在孵育4.5小时、8至9小时以及24至25小时(37℃、5%二氧化碳和95%空气)时检查精子。肝素导致精子头对头凝集,在4.5小时时使顶体上方无精液抗原的精子百分比(获能)提高20%,并使顶体反应精子的百分比增加一倍。然而,这种刺激并未提高体外受精能力。硫酸软骨素A倾向于维持活力,但不影响获能或顶体反应,可能是由于葡萄糖抑制作用。两种高渗处理都倾向于降低活力,尤其是在孵育24小时后,此时340毫渗摩尔处理的活力比295毫渗摩尔处理降低了14%。未观察到对获能的一致影响。到24小时时,340和380毫渗摩尔处理诱导的顶体反应比295毫渗摩尔处理分别多8.6%和6.1%。340毫渗摩尔/千克处理产生的体外受精率略高,但无统计学意义,这从受精卵发育到二细胞阶段可以看出。缺乏统计学意义是由于体外受精率变化较大。

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