A 32-kDa Tyrosine-phosphorylated Protein Shows a Protease-dependent Increase in Dead Boar Spermatozoa
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概要
- 論文の詳細を見る
Boar sperm TyrP32 is a 32-kDa tyrosine-phosphorylated protein that increases during the capacitation and acrosome reaction and during cryocapacitation. However, it is still unclear whether the increase in TyrP32 is an event that is limited to the process of sperm fertilization, including cryocapacitation. The aims of the present study were to demonstrate that TyrP32 is increased in dead spermatozoa after freeze-thawing without a cryoprotectant and to find the causal factors for this increase. Washed spermatozoa were resuspended in a salt solution and then frozen. The frozen samples were rapidly thawed in a warm water bath and then used for SDS-PAGE/Western blotting to detect TyrP32, SDS-PAGE/silver staining of sperm proteins and staining of acrosomal contents with FITC-PNA. In the samples before freezing, TyrP32 was barely detectable, and the distribution of the acrosomal contents was normal in most spermatozoa. One cycle of freeze-thawing induced an increase in TyrP32, a decrease in major sperm proteins and disorder in the acrosomal contents. However, the addition of a protease inhibitor (APMSF, 1 mM) suppressed the increase in TyrP32 and the decrease in the major sperm proteins, although it did not have any influence on the disorder in the acrosomal contents. Additionally, the spermatozoa did not exhibit any flagellar movement after freeze-thawing, which showed that almost all of them were dead. These results indicate that TyrP32 can show a protease-dependent increase in dead spermatozoa after freeze-thawing without a cryoprotectant even though the dead spermatozoa do not undergo cryocapacitation.
- 日本繁殖生物学会の論文
- 2008-12-01
著者
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HARAYAMA Hiroshi
Graduate School of Science and Technology, Kobe University
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Harayama Hiroshi
Graduate School Of Agricultural Sci. Kobe Univ.
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Shidara Osamu
Hyogo Prefectural Technology Center For Agriculture Forestry And Fisheries
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TABUCHI Tomohito
Graduate School of Science and Technology, Kobe University
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Tabuchi Tomohito
Graduate School Of Science And Technology Kobe University
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