Observations on the Motility of Rabbit Spermatozoa in Dilute Suspension

1. Rabbit spermatozoa suspended in Baker's solution rapidly lose motility at relatively high dilutions. At a concentration of 0.4 million per ml., the spermatozoa from most ejaculates are completely immotile within 2 or 3 hours. The same phenomenon occurs with chloride-free diluents and is therefore not due to the toxic action of chlorides. 2. This rapid immobilisation may be prevented by suspension in cell-free supernatants from other more concentrated suspensions of rabbit semen and by the accessory secretions from a vasectomised buck. The most effective supernatants are those prepared from suspensions of spermatozoa which have been left overnight before centrifuging. Immobilisation may also be prevented by many other agents, the most effective of which are gum arabic, starch, glycogen, and serum proteins (carbohydrate-containing or carbohydrate-free). Yet, in no case is the action of these substances as effective as more concentrated suspension in Baker's solution. 3. The immobilisation is not prevented by catalase, gelatin, agar, or sodium silicate. It is almost certainly not due to the toxic action of traces of heavy metals and is not affected by the use of water doubly distilled over glass in the preparation of the diluent, or by treating the diluent with activated charcoal. 4. Washing with Baker's solution does not cause immobilisation of spermatozoa suspended at 20 million per ml. at a stage at which the concentration of seminal plasma has been reduced to that equivalent to dilution to 0.4 million per ml. Further washing (six repeated centrifugings of 0.2 ml. semen in 4 ml. of Baker's solution) immobilises them. This confirms the opinion that loss of intracellular, or perhaps rather paracellular, material is a responsible agent in the dilution phenomenon.