Interdoublet sliding in bovine spermatozoa: its relationship to flagellar motility and the action of inhibitory agents.
Interdoublet sliding rates were assessed in bull sperm, utilizing a freeze-thaw procedure to allow axonemal disintegration. The sliding rate of 23 degrees C increased with increasing MgATP concentrations up to 1 mM ATP, to plateau at 8 microns/sec. The analyzed interdoublet shear in both live and demembranated (Triton X-100-extracted) bull sperm reactivated with 1 mM ATP established maximal microtubule sliding rates at 6 microns/sec during flagellar beating. Therefore, in vitro sliding rates were sufficient to account for the beat in intact flagella. The effect of inhibitors of flagellar motility on in vitro sliding rates was evaluated. While 8 microM vanadate minimally reduced the sliding rate (to approximately or equal to 4 microns/sec), only 0.5 microM vanadate was sufficient to terminate reactivated bull sperm motility. Nickel ion (0.66 mM) terminated all spontaneous motility, while only reducing microtubule sliding rates to approximately or equal to 5.0 microns/sec. Exposing intact bull sperm to theophylline (1 mM), and incubating the subsequently demembranated sperm in cAMP (3 microM), improved flagellar motility, but had little impact on microtubule sliding rates as determined by axonemal disintegration. Furthermore, deactivating live sperm with 2 mM KCN and 4 mM 2-deoxy-D-glucose renders the subsequently reactivated sperm immotile (as long as exogenous cAMP is absent). Yet, this treatment only reduced the sliding rate by 38%. Paradoxically, 4 mM MgADP reduced the sliding rates most dramatically (86%), whereas demembranated sperm models retain a strong, coordinated beating pattern in the presence of MgADP. These results demonstrate that there is no direct relationship between interdoublet sliding rates and the capacity for coordinated flagellar beating.
Bird
Z
Z
Department of Biological Sciences, Oakland University, Rochester, Michigan 48309, USA.
Hard
R
R
Kanous
K S
KS
Lindemann
C B
CB
D000244:Adenosine Diphosphate / Q000494:pharmacology; D000255:Adenosine Triphosphate / Q000378:metabolism / Q000494:pharmacology*; D000818:Animals; D002417:Cattle; D000242:Cyclic AMP / Q000494:pharmacology; D003847:Deoxyglucose / Q000494:pharmacology; D066298:In Vitro Techniques; D008297:Male; D018715:Microscopy, Video; D008870:Microtubules / Q000187:drug effects / Q000502:physiology / Q000648:ultrastructure; D008928:Mitochondria / Q000187:drug effects / Q000502:physiology / Q000648:ultrastructure; D009068:Movement; D009532:Nickel / Q000494:pharmacology; D011190:Potassium Cyanide / Q000494:pharmacology; D013081:Sperm Motility / Q000187:drug effects; D013082:Sperm Tail / Q000187:drug effects / Q000502:physiology*; D013094:Spermatozoa / Q000187:drug effects / Q000502:physiology*; D013806:Theophylline / Q000494:pharmacology; D014638:Vanadates / Q000494:pharmacology
D014638:Vanadates; C029938:nickel sulfate; D000244:Adenosine Diphosphate; D009532:Nickel; D000255:Adenosine Triphosphate; D003847:Deoxyglucose; D013806:Theophylline; D000242:Cyclic AMP; D011190:Potassium Cyanide