Identification of ejaculated proteins in the house mouse (Mus domesticus) via isotopic labeling.
Identification of ejaculated proteins in the house mouse (Mus domesticus) via isotopic labeling.
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DOI：
10.1186/1471-2164-12-306
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发表时间：
2011-06-10
期刊：
影响因子：
4.4
通讯作者：
Nachman MW
中科院分区：
文献类型：
--
作者：
Dean MD;Findlay GD;Hoopmann MR;Wu CC;MacCoss MJ;Swanson WJ;Nachman MW
精液在成功受精中起着重要作用，但对交配过程中从雄性转移到雌性的全套蛋白质的了解尚不完整。在研究最充分的哺乳动物系统之一--家鼠中，射精蛋白的清单仍然特别匮乏，在那里，人工射精技术已经被证明是不充分的。在这里，我们研究了一种识别射精蛋白的替代方法，通过用15N对雌性进行同位素标记，然后将它们与未标记的、输精管结扎的雄性交配。然后从交配的雌性中分离出蛋白质，并用质谱仪进行鉴定。除了获得对射精蛋白可能的功能和命运的洞察外，我们的研究还证明了同位素标记是研究生殖蛋白的有力手段。我们从交配后的雌性生殖道中鉴定出69种男性来源的蛋白质。在检测到的所有光谱中，超过三分之一的图谱仅映射到七个已知在交配塞子形成过程中具有重要结构意义的基因，交配塞子是一种在交配后不久形成的坚硬凝固体。精液中含有大量的蛋白质，这些蛋白质可以保护精液免受氧化应激和内肽酶的抑制。另一方面，雌性在交配时会产生内肽酶。这69个射出的蛋白质比我们之前直接从男性生殖道解剖中鉴定的其他蛋白质进化得更快。我们的研究试图全面鉴定交配过程中雄性传递给雌性的蛋白质，扩大同位素标记在哺乳动物生殖基因组学中的应用。这项技术为有针对性地监测射出的蛋白质在雌性生殖道中孵化时的命运开辟了道路。
Seminal fluid plays an important role in successful fertilization, but knowledge of the full suite of proteins transferred from males to females during copulation is incomplete. The list of ejaculated proteins remains particularly scant in one of the best-studied mammalian systems, the house mouse (Mus domesticus), where artificial ejaculation techniques have proven inadequate. Here we investigate an alternative method for identifying ejaculated proteins, by isotopically labeling females with 15N and then mating them to unlabeled, vasectomized males. Proteins were then isolated from mated females and identified using mass spectrometry. In addition to gaining insights into possible functions and fates of ejaculated proteins, our study serves as proof of concept that isotopic labeling is a powerful means to study reproductive proteins. We identified 69 male-derived proteins from the female reproductive tract following copulation. More than a third of all spectra detected mapped to just seven genes known to be structurally important in the formation of the copulatory plug, a hard coagulum that forms shortly after mating. Seminal fluid is significantly enriched for proteins that function in protection from oxidative stress and endopeptidase inhibition. Females, on the other hand, produce endopeptidases in response to mating. The 69 ejaculated proteins evolve significantly more rapidly than other proteins that we previously identified directly from dissection of the male reproductive tract. Our study attempts to comprehensively identify the proteins transferred from males to females during mating, expanding the application of isotopic labeling to mammalian reproductive genomics. This technique opens the way to the targeted monitoring of the fate of ejaculated proteins as they incubate in the female reproductive tract.
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影响因子：
56.9
作者：
DIAMOND, M
通讯作者：
DIAMOND, M
DOI：
10.1073/pnas.0603125103
发表时间：
2006-11-14
影响因子：
11.1
作者：
Aagaard, Jan E.;Yi, Xianhua;Swanson, Willie J.
通讯作者：
Swanson, Willie J.
影响因子：
64.8
作者：
DEVINE, MC
通讯作者：
DEVINE, MC
影响因子：
1.7
作者：
Carballada, R;Esponda, P
通讯作者：
Esponda, P
DOI：
10.1530/jrf.0.0790409
发表时间：
1987-03-01
期刊：
JOURNAL OF REPRODUCTION AND FERTILITY
影响因子：
--
作者：
AGRAWAL, Y;VANHAPERTTULA, T
通讯作者：
VANHAPERTTULA, T