Regulatory mechanisms of fowl sperm motility: possible role of endogenous myosin light chain kinase-like protein.
The motility of both intact and demembranated fowl spermatozoa was vigorous at 30 degrees C, but decreased markedly following the addition of 1-(5-chloronaphthalene-1-sulfonyl)-1H-hexahydro-1,4-diazepine hydrochloride (ML-9), a specific inhibitor of myosin light chain kinase (MLCK). Furthermore, the presence of a MLCK substrate peptide also inhibited the motility of demembranated spermatozoa at 30 degrees C. In contrast, the addition of N-[2-(methylamino)ethyl]-5-isoquinolinesulfonamide dihydrochloride (H-8) or N-(2-guanidinoethyl)-5-isoquinolinesulfonamide dihydrochloride (HA1004), specific inhibitors of cAMP-dependent protein kinase, did not appreciably affect the motility of either intact or demembranated spermatozoa. Cyclic AMP-dependent protein kinase substrate peptides were also ineffective for the inhibition of motility of demembranated spermatozoa at 30 degrees C. Immunoblotting of sperm extract, using an antibody to MLCK, revealed two major crossreacting proteins of 130 kDa and 61-64 kDa, which corresponded to the molecular mass of MLCK. In addition, immunogold particles, which reacted with the anti-MLCK antibody, were observed around or on the axoneme at the ultrastructural level. These results suggest that the phosphorylation of axonemal protein(s) by MLCK, or a MLCK-like protein, rather than by cAMP-dependent protein kinase, may be involved in the maintenance of fowl sperm motility at 30 degrees C.
Ashizawa
K
K
Laboratory of Animal Reproduction, Faculty of Agriculture, Miyazaki University, Japan.
Wishart
G J
GJ
Nishinakama
K
K
Sakamoto
T
T
Tsuzuki
Y
Y
D000818:Animals; D002645:Chickens / Q000378:metabolism*; D015151:Immunoblotting; D007150:Immunohistochemistry; D008297:Male; D016253:Microscopy, Immunoelectron; D009219:Myosin-Light-Chain Kinase / Q000378:metabolism*; D013081:Sperm Motility / Q000502:physiology*; D013094:Spermatozoa / Q000201:enzymology / Q000502:physiology
D009219:Myosin-Light-Chain Kinase