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KMID : 0900919930170040339
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Korean journal of Animal Reproduction
1993 Volume.17 No. 4 p.339 ~ p.346
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Isolation of Bovine Spermatozoal Components by Physical or Chemical Treatments
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Lee Sang-Ho
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Abstract
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An understanding of the structure and function of mammalian spermatozoa requires the iso-lation of these components. In this study, frozen-thawed bovine spermatozoa were treated by physical treatments (vortexing, 26 gauge needle, strained 26 gauge needles and freezing-thawing) or chemical treatments (trypsin, dithiothreitol, sodium dodecylsulfate and beta-mercaptoethanoJ) to yield free heads and tails. The most effective treatment was repeated pumping of sperm suspension through a strained 26 gauge needle conneted to a syringe. Spermatozoa by this treatment were mainly broken at the junction of the head and the tail, resulting in 90-100% yields. Also, sperm head surface did not modify during strained 26 gauge needle treatment when either spermatozoa or sperm heads were incubated in 250{mu}textrm{g}/ml of FITC-UEA 1 for 1 h at room temperature to detect the modification of sperm surface components. Other physical treatments were less efficient for the breakdown of spermatozoa. The effects of chemical treatments on bovine spermatozoa are not noticeable. Dissected sperm heads and tails should be fractional leading to nearly pure components by sucrose gradient centrifugation at 1,000 rpm for 15 min. The result suggest that the established method may be useful for the biochemical study of spermatozoal components, and the understanding of oocyte activation mechanism either by spermatozoal components during fertilization or microinjection of isolated components.
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KEYWORD
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physical dissection, chemical dissection, bovine spermatozoa
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