A monoclonal antibody against the dynein IC1 peptide of sea urchin spermatozoa inhibits the motility of sea urchin, dinoflagellate, and human flagellar axonemes.
To investigate the role of axonemal components in the mechanics and regulation of flagellar movement, we have generated a series of monoclonal antibodies (mAb) against sea urchin (Lytechinus pictus) sperm axonemal proteins, selected for their ability to inhibit the motility of demembranated sperm models. One of these antibodies, mAb D1, recognizes an antigen of 142 kDa on blots of sea urchin axonemal proteins and of purified outer arm dynein, suggesting that it acts by binding to the heaviest intermediate chain (IC1) of the dynein arm. mAb D1 blocks the motility of demembranated sea urchin spermatozoa by modifying the beating amplitude and shear angle without affecting the ATPase activity of purified dynein or of demembranated immotile spermatozoa. Furthermore, mAb D1 had only a marginal effect on the velocity of sliding microtubules in trypsin-treated axonemes. This antibody was also capable of inhibiting the motility of flagella of Oxyrrhis marina, a primitive dinoflagellate, and those of demembranated human spermatozoa. Localization of the antigen recognized by mAb D1 by immunofluorescence reveals its presence on the axonemes of flagella from sea urchin spermatozoa and O. marina but not on the cortical microtubule network of the dinoflagellate. These results are consistent with a dynamic role for the dynein intermediate chain IC1 in the bending and/or wave propagation of flagellar axonemes.
Urology Research Laboratory, Royal Victoria Hospital, Faculty of Medicine, McGill University, Montreal, Canada.
C
C
Gagnon
D
D
White
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P
Huitorel
J
J
Cosson
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000494:pharmacology*; D000918:Antibody Specificity; D004141:Dinoflagellida / Q000276:immunology*; D004398:Dyneins / Q000276:immunology*; D006801:Humans; D008297:Male; D010446:Peptide Fragments / Q000276:immunology*; D012617:Sea Urchins / Q000276:immunology*; D013045:Species Specificity; D013081:Sperm Motility / Q000187:drug effects*; D013082:Sperm Tail / Q000737:chemistry / Q000502:physiology*; D013094:Spermatozoa / Q000737:chemistry / Q000276:immunology / Q000502:physiology*
D000911:Antibodies, Monoclonal; D010446:Peptide Fragments; D000251:Adenosine Triphosphatases; D004398:Dyneins