Fertilization acid release in Urechis eggs. I. The nature of the acid and the dependence of acid release and egg activation on external pH.

The amount of fertilization acid produced by eggs of Urechis caupo , monitored by automatically back-titrating egg suspensions with base, depends linearly on the pH of the seawater. Above pH 7.0, at which no acid is released (Paul, M., Dev. Biol. 43, 299-312, 1975), acid release increased approximately 0.34 pmole/egg/0.1 pH unit. Activation (germinal vesicle breakdown) depended on the amount of acid release in natural seawater; it did not occur if eggs released less than 1.5 pmole acid/egg. When fertilization acid is released into HCO-3-free seawater and the pH permitted to decrease, the supernatant can be tested for the presence of a volatile acid, such as CO2, by bubbling with N2 and comparing the increase in pH as volatile acid is driven off with experiments in which HCl or CO2 is substituted for fertilization acid. An increase in pH of less than 0.2 pH units occurred on N2 bubbling when fertilization acid or HCl was used to acidify HCO-3-free seawater compared to an increase of greater than 0.5 pH units when CO2 was used. Therefore, most, if not all, of Urechis fertilization acid is not volatile, and since Paul (1975) showed that it is not a nonvolatile weak acid, it must be H+.