Functional deficiencies and a reduced response to calcium in the flagellum of mouse sperm lacking SPAG16L.
The Spag16L gene codes for a protein that is localized to the central apparatus which is essential for normal sperm motility and male fertility. Sperm from mice homozygous for a targeted deletion of the Spag16L gene were examined to assess their flagellar motor functions compared with age- and strain-matched control sperm. Sperm were also demembranated with Triton X-100 and examined for their ability to respond to free calcium, as well as for their ability to undergo microtubule sliding driven by dynein action. In addition, the passive flagella, inhibited by sodium metavanadate to disable the dyneins, were examined for mechanical abnormalities. Live Spag16L-null sperm exhibited much less bending of the flagellum during the beat. The amount of microtubule sliding in the R-bend direction of the beat was selectively restricted, which suggests that there is limited activation of the dyneins on one side of the axoneme in the live cells. This is corroborated by the results on detergent-extracted sperm models. The flagellar response to calcium is greatly reduced. The calcium response requires the activation of the dyneins on outer doublets 1, 2, 3, and 4. These are the same dyneins required for R-bend formation. In axonemes prepared to disintegrate by microtubule sliding, we observed little or no extrusion of doublets 1 and 2, consistent with a reduced activity of their dyneins. This deficit in motor function, and an increased rigidity of the midpiece region which we detected in the passive flagella, together can explain the observed motility characteristics of the Spag16L-null sperm.
Lesich
Kathleen A
KA
Department of Biological Sciences, Oakland University, Rochester, Michigan, USA.
Zhang
Zhibing
Z
Kelsch
Courtney B
CB
Ponichter
Kristen L
KL
Strauss
Jerome F
JF
Lindemann
Charles B
CB
D000818:Animals; D002118:Calcium / Q000494:pharmacology*; D015536:Down-Regulation / Q000187:drug effects; D004548:Elasticity / Q000502:physiology; D007248:Infertility, Male / Q000235:genetics* / Q000378:metabolism / Q000473:pathology / Q000503:physiopathology; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D008869:Microtubule-Associated Proteins / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D032961:Sperm Midpiece / Q000187:drug effects / Q000378:metabolism / Q000473:pathology / Q000502:physiology; D013081:Sperm Motility / Q000187:drug effects / Q000235:genetics* / Q000502:physiology; D013082:Sperm Tail / Q000187:drug effects* / Q000378:metabolism / Q000502:physiology / Q000648:ultrastructure; D013094:Spermatozoa / Q000187:drug effects / Q000378:metabolism / Q000473:pathology / Q000502:physiology
D008869:Microtubule-Associated Proteins; C491467:PF20 protein, mouse; D002118:Calcium