Characteristics of sperm acrosin-like activity of paddlefish (Polyodon spathula Walbaum).
Spermatozoa of paddlefish and sturgeon fishes (Acipenseriformes), unlike teleost fish, have an acrosome. The objectives of this study were to characterize acrosin-like activity of cryopreserved sperm of paddlefish (Polyodon spathula) and to test and compare stability of paddlefish acrosin-like activity with that of lake sturgeon and bull spermatozoa. Mean acrosin-like activity of cryopreserved paddlefish sperm was 0.372 +/- 0.067 microU/10(6) spermatozoa. This activity was 79% higher in the whole semen than in spermatozoa. Highest activity was recorded at pH 8.0 and 8.5. Triton X-100, zinc ions and 4'-acetamidophenyl 4-guanidinobenzoate (AGB) inhibited the activity. Amidase activity was also inhibited by N-alpha-p-tosyl-L-lysine chloromethyl ketone (TLCK) and N-tosyl-L-phenylalanine chloromethyl ketone (TPCK). TLCK at concentrations of 0.1 and 1.0 mM gave a significant decrease in activity of 19 and 61%, respectively. However, TPCK significantly inhibited amidase activity (by 19%) only at concentration 1.0 mM. After acidification and 60 min incubation at 4 degrees C of sperm suspensions only 4% of the activity was retained. A similar phenomenon was observed in the case of lake sturgeon but not bull sperm. These results suggest that trypsin-like activity of Acipenserid fish resembles rather fish trypsin that mammalian one. In frozen-thawed paddlefish sperm a minute chymotrypsin-like activity was also indicated, when GPNA was used as substrate. This activity amounted to 0.0415 +/- 0.0138 microU/10(6) spermatozoa and was 18% of total amidase activity. This suggests that chymotrypsin-like activity may also be present in paddlefish spermatozoa.
Ciereszko
A
A
Department of Molecular Andrology, Polish Academy of Sciences, Olsztyn, Poland. acieresz@food.irzbz.pan.olsztyn.pl
Dabrowski
K
K
Mims
S D
SD
Glogowski
J
J
D000176:Acrosin / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D000177:Acrosome / Q000201:enzymology*; D000818:Animals; D002417:Cattle; D015925:Cryopreservation; D003902:Detergents / Q000494:pharmacology; D004795:Enzyme Stability; D005399:Fishes; D006863:Hydrogen-Ion Concentration; D008297:Male; D012661:Semen / Q000201:enzymology; D012662:Semen Preservation; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology; D013094:Spermatozoa / Q000187:drug effects / Q000201:enzymology*; D015032:Zinc / Q000494:pharmacology
D003902:Detergents; D015842:Serine Proteinase Inhibitors; D000176:Acrosin; D015032:Zinc