Bench G, Corzett M H, De Yebra L, Oliva R, Balhorn R
Center for Accelerator Mass Spectrometry, Lawrence Livermore National Laboratory, Livermore, CA 94550, USA.
Mol Reprod Dev. 1998 Jul;50(3):345-53. doi: 10.1002/(SICI)1098-2795(199807)50:3<345::AID-MRD11>3.0.CO;2-3.
Sperm from 2 semen samples collected 6 months apart from an infertile male and 3 semen samples collected over an 18-month period from a fertile human male volunteer have been analyzed for their protamine and DNA content. Hup1M and Hup2b antibodies were used to detect the presence of protamines and protamine precursors in western blots of nuclear proteins isolated from pools of sperm. Phosphorus and sulfur contents, which can be used to estimate the nuclear DNA and protamine contents of sperm from fertile males, were measured within individual sperm heads from each semen sample by particle induced x-ray emission (PIXE). The single-cell data reveal no significant differences in the phosphorus and sulfur contents of sperm heads in the three semen samples obtained from the fertile male. For the initial semen sample produced by the infertile male, Western blot data show a normal complement of protamine 1, small amounts of mature protamine 2, and reveal large amounts of anti-protamine 2 reactive proteins with electrophoretic mobilities similar to protamine 2 precursors. Data from PIXE show elevated levels of sulfur within sperm heads compared with sperm from the fertile male. Western blot data exhibit no evidence of protamines or protamine 2 precursors in the second semen sample produced by the infertile male. Data from PIXE suggest that these sperm are highly deficient in sulfur and protamines. These results show that the degree of maturation of sperm cells present in the semen of some infertile males can vary with time.
对一名不育男性间隔6个月采集的2份精液样本以及一名生育能力正常的男性志愿者在18个月内采集的3份精液样本中的精子进行了鱼精蛋白和DNA含量分析。使用Hup1M和Hup2b抗体，通过从精子库中分离出的核蛋白的蛋白质免疫印迹法检测鱼精蛋白和鱼精蛋白前体的存在。通过粒子诱导X射线发射（PIXE）测量了每个精液样本中单个精子头部的磷和硫含量，这些含量可用于估计生育能力正常男性精子的核DNA和鱼精蛋白含量。单细胞数据显示，从生育能力正常的男性获得的3份精液样本中，精子头部的磷和硫含量没有显著差异。对于不育男性产生的初始精液样本，蛋白质免疫印迹数据显示鱼精蛋白1的含量正常，有少量成熟的鱼精蛋白2，并显示出大量与鱼精蛋白2前体电泳迁移率相似的抗鱼精蛋白2反应性蛋白。PIXE数据显示，与生育能力正常男性的精子相比，不育男性精子头部的硫含量升高。蛋白质免疫印迹数据显示，不育男性产生的第二个精液样本中没有鱼精蛋白或鱼精蛋白2前体的证据。PIXE数据表明，这些精子的硫和鱼精蛋白严重缺乏。这些结果表明，一些不育男性精液中存在的精子细胞成熟程度可能随时间而变化。
