Methods for quantitating sea urchin sperm-egg binding.
Methods for quantitating sea urchin sperm-egg binding.
复制标题
定量海胆精卵结合的方法。
DOI：
10.1016/0014-4827(73)90265-6
复制
发表时间：
1973
影响因子：
3.7
通讯作者：
Judith E. Payne
中科院分区：
文献类型：
--
作者：
V. Vacquier;Judith E. Payne
Two simple photometric methods are described for determining the average number of bound sea urchin spermatozoa per egg between 0 and 60 sec after insemination. One method is based on stopping gamete interaction with formaldehyde and then, after the eggs settle with sperm bound to their surfaces, measuring the turbidity of sperm remaining in suspension. An alternative method involves removal of the dead, unbound sperm from the formaldehyde fixed eggs by repeated washing in sea water. The bound sperm are then released from the egg surface by pronase digestion and the turbidity of the sperm suspension measured and related to sperm concentration by direct cell counts. Two phases of gamete interaction exist (1) the binding phase, from 0 to 20 or 25 sec, during which time sperm continuously bind to the egg surfaces; (2) the unbinding phase, from 20 or 25 to 50 sec, during which time the sperm are unbound from the eggs and return to the suspension. The results of experiments in which the method is used to assess sperm binding at different pH values and at different calcium concentrations are presented. Data are presented suggesting that a definite number of sperm binding sites may exist on the vitelline layer.
