Ishijima S A, Okuno M, Odagiri H, Mohri T, Mohri H
Department of Biology, College of Arts and Sciences, University of Tokyo, Japan.
Zoolog Sci. 1992 Jun;9(3):601-6.
The effectiveness of separation of murine X- and Y-bearing sperm by free-flow electrophoresis was evaluated by the polymerase chain reaction (PCR). The ratio of X- and Y-bearing sperm from cauda epididymis was analyzed before and after free-flow electrophoresis. A Y-chromosome-specific sequence (pY353/B) and an autosomal sequence (myogenin) were used to estimate the ratio between X- and Y-sperm in the separated fractions. Cauda epididymal mice sperm were separated into two peak fractions under the electric field. Each peak fraction contained sperm of normal shape, however, the motility of the sperm was extremely diminished after separation by electrophoresis. DNA was extracted from 10(4) sperm from each fraction and from the unseparated sperm, and Y-chromosome specific PCR was performed. The PCR experiment revealed that fraction No. 16 (the peak near the cathode) was a Y-sperm rich fraction, whereas fraction No. 22 (the peak near the anode) was a Y-sperm poor one. These results suggested that murine X- and Y-sperm could be successfully separated by free-flow electrophoresis. Analysis of the chromosome-specific sequence by PCR was demonstrated to be a direct and adequate method to evaluate the separation of X- and Y-sperm.
通过聚合酶链反应（PCR）评估了自由流动电泳分离小鼠X和Y精子的有效性。分析了自由流动电泳前后附睾尾部X和Y精子的比例。使用Y染色体特异性序列（pY353/B）和常染色体序列（肌细胞生成素）来估计分离组分中X和Y精子之间的比例。附睾尾部小鼠精子在电场作用下被分离成两个峰值组分。每个峰值组分都含有形状正常的精子，然而，电泳分离后精子的活力极度下降。从每个组分的10⁴个精子以及未分离的精子中提取DNA，并进行Y染色体特异性PCR。PCR实验表明，第16号组分（阴极附近的峰值）是富含Y精子的组分，而第22号组分（阳极附近的峰值）是Y精子含量低的组分。这些结果表明，小鼠X和Y精子可以通过自由流动电泳成功分离。通过PCR分析染色体特异性序列被证明是评估X和Y精子分离的一种直接且合适的方法。
