Effects of tyrosine kinase inhibitor on the motility and ATP concentrations of fowl spermatozoa.
The possible role of tyrosine kinase in the regulation of fowl sperm motility was investigated by using a stable analogue of erbstatin, methyl 2,5-dihydroxycinnamate (2,5-MeC), a specific inhibitor of tyrosine kinase. This inhibited the motility of intact spermatozoa at 30 degrees C in a dose-dependent manner. In contrast, the motility of demembranated spermatozoa was not inhibited by the same concentrations of 2,5-MeC. At 40 degrees C, both intact and demembranated spermatozoa were almost immotile with or without 2,5-MeC. Additionally, intact spermatozoa, stimulated by the addition of Ca2+ or calyculin A, a specific inhibitor of protein phosphatases, lost their motility with the subsequent addition of 2,5-MeC at 40 degrees C. However, unlike the motility, the ATP concentrations of spermatozoa were maintained in about 30-35 nmol ATP/10(9) cells during these incubation periods. The activity of tyrosine kinase of spermatozoa at 30 degrees C, estimated by measuring the phosphorylation of a synthetic peptide substrate, RR-SRC, was 0.17 pmol/min per milligram of protein. This activity was lower than that of fowl testes or chick brain but higher than that of chick liver. These results suggest that tyrosine kinase activity, which is not retained in the axoneme and/or accessory cytoskeletal components, may be involved in the maintenance of flagellar movement of fowl spermatozoa at 30 degrees C.
Ashizawa
K
K
Laboratory of Animal Reproduction, Faculty of Agriculture, Miyazaki University, Japan. a0c303u@cc.miyazaki-u.ac.jp
Higashio
M
M
Tsuzuki
Y
Y
D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D002462:Cell Membrane / Q000502:physiology; D002645:Chickens; D002934:Cinnamates / Q000494:pharmacology; D004789:Enzyme Activation; D008297:Male; D009928:Organ Specificity; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D013081:Sperm Motility / Q000187:drug effects*; D013094:Spermatozoa / Q000201:enzymology* / Q000378:metabolism / Q000502:physiology
D002934:Cinnamates; C087931:ethyl 2,5-dihydroxycinnamate; D000255:Adenosine Triphosphate; D011505:Protein-Tyrosine Kinases