Ejaculated and Epididymal Mouse Spermatozoa Are Different in Their Susceptibility to Nuclease-Dependent DNA Damage and in Their Nuclease Activity1
Ejaculated and Epididymal Mouse Spermatozoa Are Different in Their Susceptibility to Nuclease-Dependent DNA Damage and in Their Nuclease Activity1
复制标题
小鼠射精精子和附睾精子对核酸酶依赖性 DNA 损伤的敏感性及其核酸酶活性不同1
作者：
Y. Yamauchi;A. Ajduk;Jonathan M. Riel;M. Ward
摘要从子宫中取出的射精精子在冷冻干燥和无冷冻保护的冷冻过程中比附睾精子更容易受到DNA损伤。这促使我们推测，交配后子宫中存在的一个因素，无论是男性还是女性，都是导致射出的精子在保存过程中对DNA损伤的敏感性增加的原因，而附睾和射出的小鼠精子对应激反应的差异源于不同的核酸酶活性。我们首先将附睾精子暴露于输精管结扎组(UCSP)的子宫内容物(UCSP)、发情期未交配的雌性(UC)子宫内容物(UC)和精囊液(SVF)中，并在卵胞浆内单精子注射(ICSI)后对精子染色体进行了检查。我们发现，暴露于UCSP和SVF后，精子染色体断裂发生率增加，DNA断裂非常严重，但暴露于UC的精子染色体正常。彗星试验结果证实，受精前精子中存在暴露于SVF后的DNA损伤。接下来，我们用质粒消化试验检测了精子及其相关成分中的核酸酶活性。核酸酶活性在分离的附睾液和射精精子、附睾液和精浆中均可检测到，且在所有来自射精液的样品中表现得更为明显。本研究的综合结果表明，附睾和射出的小鼠精子标本对核酸酶依赖的DNA损伤的敏感性存在内在差异，这种损伤源于它们的核酸酶活性。这种核酸酶活性在制剂的无精子部分和分离的精子中都被检测到。
Abstract Ejaculated mouse sperm retrieved from the uteri are more susceptible to DNA damage during freeze-drying and freezing without cryoprotection than epididymal sperm. This prompted us to speculate that a factor present in the uterus after mating, either male or female derived, was responsible for increased susceptibility of ejaculated sperm to DNA damage during preservation and that the differences between epididymal and ejaculated mouse sperm in response to stress originated from varying nuclease activity. We first exposed epididymal sperm to the uterine content from females mated to vasectomized males (UCSP), to the uterine content from unmated females in estrus (UC), and to the seminal vesicle fluid (SVF) and examined sperm chromosomes after intracytoplasmic sperm injection (ICSI). We found an increased incidence of chromosome breaks and extremely severe DNA breakage after exposure to UCSP and SVF, respectively, but the chromosomes were normal in sperm exposed to UC. Comet assay results verified that DNA damage after exposure to SVF was present in sperm before fertilization. Next, we examined nuclease activity in sperm and their associated components with a plasmid digestion assay. Nuclease activity was detected in isolated epididymal and ejaculated sperm, as well as in epididymal fluid and seminal plasma, and was much more pronounced in all samples originating from ejaculate. The combined results from the present study imply that there are intrinsic differences between the epididymal and ejaculated mouse sperm preparations in their susceptibility to nuclease-dependent DNA damage that originates from their nuclease activity. This nuclease activity was detected both in the sperm-free fraction of preparations and isolated sperm.
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影响因子：
3.6
作者：
KIMURA, Y;YANAGIMACHI, R
通讯作者：
YANAGIMACHI, R
DOI：
10.1071/rd03072
发表时间：
2004-07
期刊：
Reproduction, fertility, and development
影响因子：
--
作者：
M. Ward;W. Ward
通讯作者：
M. Ward;W. Ward
影响因子：
3.6
作者：
Kuretake, S;Kimura, Y;Yanagimachi, R
通讯作者：
Yanagimachi, R
影响因子：
--
作者：
Barbara Sotolongo;Thomas T F Huang;Elisabete Isenberger;W. Ward
通讯作者：
Barbara Sotolongo;Thomas T F Huang;Elisabete Isenberger;W. Ward
DOI：
10.1530/jrf.0.1120011
发表时间：
1998-01-01
期刊：
JOURNAL OF REPRODUCTION AND FERTILITY
影响因子：
--
作者：
Wakayama, T;Whittingham, DG;Yanagimachi, R
通讯作者：
Yanagimachi, R