The Physiology of Sea-Urchin Spermatozoa

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Abstract
1. Spermatozoa and seminal plasma of Echinus esculentus contain catalase. 2. At 15° C., 4 ml. of a suspension of semen diluted with neutral phosphate buffer in the ratio 1:13 produced in 1 min. 90µl. O2 from an H2O2 solution containing 150 µl. O2. The dry weight of semen in the suspension was 45 mg. and the number of spermatozoa 8.55x109. Under the same conditions, seminal plasma obtained by centrifuging semen produced 50 µl. O2 in 1 min. The dry weight of seminal plasma in the suspension was 12 mg. Human blood, dry weight 229.3 mg./ml., must be diluted with phosphate buffer in the ratio 1:1700 to produce the same amount of O2 in 1 min. as the above suspension of semen. If catalatic activity is defined by the equation Ac = (gt)-1 In {a/(a-x)}, where g = weight in g./ml. of the catalase-containing material, t = 1 min., a = initial substrate concentration (H2O2), and x = amount of H2O2 decomposed in 1 min. at 15° C., Ac = 80-100, 150-200 and 6800 respectively for sea-urchin semen, sea-urchin seminal plasma and human blood. 3. The catalatic activity of semen and seminal plasma is strongly inhibited by hydroxylamine. 4. The O2 uptake and motility of sea-urchin spermatozoa is unaffected by M/5000 H2O2. Higher concentrations of H2O2, M/3000-5000, produce a pronounced ‘shock’ effect, from which the spermatozoa often completely recover. 5. Low concentrations of hydroxylamine, M/3000, reduce O2 uptake and motility. 6. Sea-urchin spermatozoa are almost instantly killed by combinations of hydroxylamine and H2O2, at concentrations which are relatively innocuous when the substances are added separately. 7. A rough calculation indicates that a single spermatozoon contains less than 500 molecules of catalase. 8. A new method of adding H2O2 to catalase-containing material in a manometer is described.