ウニ精子の分画

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As a step in studies on the mechanism of spermatozoan motility, it has been tried to fractionate sperm cells into head, mid-piece and tail as cleanly as possible. Spermatozoa of sea urchins, Anthocidaris crassispina, Hemicentrotus pulcherrimus and Pseudocentrotus depressus, were washed with Mg^<++> and Ca^<++>-free artificial sea water and homogenized with a Teflon homogenizer. The homogenate was layered on 0.75M sucrose solution and centrifuged. By this treatment, tail fraction was isolated from the fraction containing head and mid-piece. The latter fraction was then treated with trypsin (2.5μg/ml) and again homogenized. The homogenate thus obtained was layered on 1.5M sucrose solution and centrifuged, and separated into head and mid-piece fractions (Fig. 1). The purity of the three fractions was tested by measuring DNA content and cytochrome oxidase activity and by observation under an electron microscope. As shown in Table 1, the separation into head or mid-piece (mitochondria) fractions appeared to be almost complete. Electron micrograph of the tail fraction also revealed that the fraction contained only tails without any significant contamination.
社団法人日本動物学会の論文
1969-12-15