As the effect of carbon dioxide on mammalian spermatozoa is of current interest (Lodge & Salisbury, 1963; Hammer & Williams, 1964; VanDemark, Koyama & Lodge, 1965; Wales & O'Shea, 1966; Wales & Restall, 1966), the possibility of its fixation by ram spermatozoa has been investigated.
Aliquots (0·4 ml) of ram semen or twice-washed spermatozoa (5·4 × 108 cells/ flask) were incubated at 37° C with [14C]sodium bicarbonate (1 μmole and 25 μc) in phosphate-buffered saline containing 1 mm-potassium chloride, 2mm-magnesium chloride, 10 mm-sodium Pyruvate, 30 mg/100 ml penicillin and 50 mg/100 ml streptomycin. After 15 or 60 min, 0·5 ml of the incubated semen was removed and stored at -15° C for subsequent chromatography. The remaining 0·5 ml was acidified (0·1 ml of 6 n-H2
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