The reactivation of demembranated human spermatozoa lacking outer dynein arms is independent of pH.
The effect of pH, Mg-ATP, and free calcium on activity of the inner dynein arm was investigated using demembranated human spermatozoa lacking the outer dynein arms (LODA). The results were compared with those obtained for demembranated-reactivated normal spermatozoa to evaluate the functional properties of the inner and outer dynein arms in axonemal motility. The reactivation of Triton X-100-demembranated LODA spermatozoa was analysed at various pHs and concentrations of Mg-ATP and calcium using video recordings. The percentage of reactivated LODA spermatozoa as a function of Mg-ATP concentration was not dependent on pH, whereas reactivation of normal human spermatozoa is pH dependent. This suggests that there may be a pH-dependent regulatory mechanism associated with the outer dynein arms. A delay in the principal bend propagation of normal and LODA reactivated cells was found at pH 7.1. This disappeared at pH 7.8 in normal but not in LODA populations. This suggests a role for outer dynein arms in the initiation of the propagation of flagellar bends at alkaline pH. The level of LODA and normal sperm reactivation both depended on the calcium concentration in the medium. At lower free calcium concentrations, the reactivation level and beat frequency of reactivated cells were higher. Our results suggest a functional difference between outer and inner dynein arms of human spermatozoa based on a differential pH sensitivity. Moreover, calcium seems to exert its regulatory action elsewhere than on the outer dynein arms.
Keskes
L
L
Laboratoire de Biologie de la Reproduction, Université Paris V, Hopital Cochin-Port-Royal, France.
Giroux-Widemann
V
V
Serres
C
C
Pignot-Paintrand
I
I
Jouannet
P
P
Feneux
D
D
D000255:Adenosine Triphosphate / Q000494:pharmacology; D002118:Calcium / Q000494:pharmacology; D002462:Cell Membrane / Q000378:metabolism / Q000473:pathology / Q000648:ultrastructure; D002614:Chelating Agents / Q000378:metabolism; D004398:Dyneins / Q000172:deficiency*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008297:Male; D008854:Microscopy, Electron; D013076:Sperm Count / Q000187:drug effects; D013081:Sperm Motility / Q000187:drug effects; D013094:Spermatozoa / Q000378:metabolism* / Q000473:pathology* / Q000648:ultrastructure
D002614:Chelating Agents; D000255:Adenosine Triphosphate; D004398:Dyneins; D002118:Calcium