Effects of medium containing heparin and theophylline on capacitation and metabolic enzyme activities of ejaculated spermatozoa from dogs with asthenozoospermia

The percentages of motile sperm (%MO), hyperactivated sperm (%HA), and acro some-reacted sperm (%AR) of four beagle dogs with asthenozoospermia (AS) an d five normal beagle dogs were determined during 7 h of incubation. The met abolic enzyme activities of the sperm was examined after 0 and 4 h of incub ation. The sperm were incubated in canine capacitation medium (CCM) and CCM containing either 20 mu g ml(-1) heparin (HE), 10 mu g ml(-1) theophylline (TH) or 20 mu g ml(-1) HE + 10 mu g ml(-1) TH in glass tubes at 38 degrees C under 5% CO2 in air. The %HA and %AR were determined by counting the sper m exhibiting star-spin like movement and by the triple stain technique. The spermatozoa in HE + TH CCM were homogenized and centrifuged, and the metab olic activities of hexokinase, fructokinase, glucose-6-phosphodehydrogenase (G6PD), and pyruvate kinase in the sperm cytosol in the supernatant was me asured with a spectrophotometer. The mean %MO and %HA values of both AS and normal does in the four types of CCM were highest in HE + TH CCM; with a m ean %HA in HE + TH CCM of 78 +/- 5% (S.E.) after 7 h oi incubation. However , there was little difference in %AR among the four types of CCM. The mean activities of the four enzymes in the sperm of AS dogs before incubation wa s significantly lower than in the sperm of normal dogs (P < 0.05, 0.01). Ho wever, after 4 h of incubation the activities of all enzymes in the sperm o f both AS and normal dogs was clearly higher in HE + TH CCM than in the con trol CCM. These findings indicate that HE and TH in the medium are effectiv e in activating metabolic enzymes. maintaining longer sperm motility, and e fficiently inducing HA even of the sperm of AS dogs. (C)1999 Elsevier Scien ce B.V. All rights reserved.