Abstract
ABSTRACT An investigation has been made into the causes of the interrelated phenomena of Senescence and the Dilution Effect in the spermatozoa of Echinus esculentus. The Dilution Effect is the increase in O2 uptake and in movement observed when a concentrated sperm suspension, obtained by partial dilution of semen, is diluted with sea water. The supernatant sea water obtained by centrifuging partially metabolized sperm suspensions contains no substances which depress the respiration or motility of the spermatozoa of the same species. This supernatant sea water has a small but definite protective action on spermatozoa in maintaining but not increasing their O2 consumption. The effect is enhanced by agitating the suspensions before centrifugation, E. esculentus spermatozoa do not glycolyse aerobically, anaerobically, or in the presence of 5% O2 in CO. The absence of aerobic acid production proves that the reduced O2 uptake per sperm of dense suspensions is not due to a lowering of the pH of the medium. As the Dilution Effect is observed in manometric experiments in which CO2 is continuously and adequately removed, neither accumulation of CO2 nor a reduction in pH resulting from the accumulation of CO2 are responsible for the reduced O2 uptake of dense suspensions. The O2 uptake of dense sperm suspensions ( > 109 sperm/ml.) is altered by changes in O2 tension. O2 consumption in 100% oxygen is nearly double that in 10% O2 in Ns. An increase in O2 tension has no stimulating action on dilute suspensions. An atmosphere of 100% O2 has a gradual toxic effect on dense and dilute sperm suspensions. Carbon monoxide has a greater inhibitory action on dilute than on concentrated sperm suspensions. The difference between the effects of varying O2 tensions or CO on dense and dilute suspensions is partly, or perhaps wholly, due to gaseous diffusion being a limiting factor. This may partly explain the Dilution Effect. Apart from photo-reversibly inhibiting respiration, CO exerts an irreversible toxic influence on sea-urchin spermatozoa. Illumination of spermatozoa with a wave-length of light outside the region of photo-chemical absorption by CO cytochrome oxidase enables visual examination to be made in the presence of CO. The O2 uptake of a suitable concentration of spermatozoa is 70% inhibited by 5% O2 in CO. The same inhibition occurs when the suspension is illuminated by light of wave-length 548 mμ, but there is no equivalent decrease in motility. After motility and respiration have ceased sea-urchin spermatozoa still contain oxidizable substrates and the appropriate dehydrogenases as evidenced by their ability to reduce methylene blue in an evacuated Thunberg tube.
Published Version
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