Equilibration in freezing extender alters in vitro sperm-oviduct binding in the domestic cat (Felis catus).

For the preservation of endangered felid species, epididymal sperm may be received from valuable individuals after castration or death and they need to be cryopreserved for storage. However, pregnancy rates with epididymal or cryopreserved sperm are lower than with ejaculated and non-frozen semen even if insemination is surgically performed into the oviduct. To investigate whether equilibration, the first step of the cryopreservation procedure, has an impact on sperm-oviduct binding, we generated oviduct epithelial cell vesicles from isthmus segments of preovulatory domestic cats. Binding assays were performed with epididymal sperm in a cell culture medium (M199) without supplements, or after cooling to 15 degrees C in a freezing extender (TestG), supplemented with glycerol and the water-soluble fraction of hen's egg yolk mainly comprising LDL. The sperm-oviduct binding was assessed both quantitatively and qualitatively (head or tail binding of sperm with active or inactive mitochondria). Most of the bound sperm prepared in M199 had active mitochondria and were attached to the vesicles by their heads. In equilibrated samples, the proportion of bound sperm with active mitochondria and the proportion of head-bound spermatozoa were reduced. The total motility of the sperm after 1 h of incubation in the absence or presence of vesicles were also affected by the preparation (higher in equilibrated) and the incubation (lower in co-incubated), while mitochondrial activity was influenced just by the preparation. Obviously, LDL has a beneficial effect on sperm motility, but we suggest that it interferes with the molecular sperm-oviduct crosstalk and causes a reduced binding of "good" sperm.