Tetramethylrhodamine-labelled microtubules were observed to move on rows of dynein arms of sea urchin sperm axonemes exposed by elastase-induced sliding disintegration. The microtubules moved towards the flagellar tip at a velocity of 3.1+/−2.1 microm second-1 (mean +/− s.d., n=53) in the presence of 0.1 mM ATP at 22 degrees C, but none moved towards the sperm head. We also examined the polarity of microtubule binding to axonemal doublet microtubules in the absence of ATP by using microtubules brightly labelled at their minus-ends. In 140 of 210 microtubules studied, they bound to axonemal microtubules with a parallel polarity. These results suggest that tightly packed dynein arms on the outer doublet microtubules of sperm axoneme preferentially bind microtubules to themselves with the same polarity as that of the axoneme and that they generate a force to move only these microtubules in the direction away from the sperm head.

REFERENCES

Brokaw
C. J.
(
1966
).
Effects of increased viscosity on the movements of some invertebrate spermatozoa.
J. Exp. Biol
45
,
113
139
Brokaw
C. J.
(
1980
).
Elastase digestion of demembranated sperm flagella.
Science
207
,
1365
1367
Fox
L. A.
,
Sale
W. S.
(
1987
).
Direction of force generated by the inner row of dynein arms on flagellar microtubules.
J. Cell Biol
105
,
1781
1787
Gibbons
B. H.
,
Gibbons
I. R.
(
1972
).
Flagellar movement and adenosine triphosphatase activity in sea urchin sperm extracted with Triton X-100.
J. Cell Biol
54
,
75
97
Gibbons
I. R.
(
1981
).
Cilia and flagella of eukaryotes.
J. Cell Biol
91
,
107
–.
Goodenough
U. W.
,
Heuser
J. E.
(
1982
).
Substructure of the outer dynein arm.
J. Cell Biol
95
,
798
815
Goodenough
U. W.
,
Heuser
J. E.
(
1985
).
Substructure of inner dynein arms, radial spokes, and the central pair/projection complex of cilia and flagella.
J. Cell Biol
100
,
2008
2018
Howard
J.
,
Hyman
A. A.
(
1993
).
Preparation of marked microtubules for the assay of the polarity of microtubule-based motors by fluorescence microscopy.
Meth. Cell Biol
39
,
105
113
Hyman
A.
,
Drechsel
D.
,
Kellogg
D.
,
Salser
S.
,
Sawin
K.
,
Steffen
P.
,
Wordeman
L.
,
Mitchison
T.
(
1991
).
Preparation of modified tubulins.
Meth. Enzymol
196
,
478
485
Ishijima
S.
,
Kubo-Irie
M.
,
Mohri
H.
,
Hamaguchi
Y.
(
1996
).
Calcium-dependent bidirectional power stroke of the dynein arms in sea urchin sperm axonemes.
J. Cell Sci
109
,
2833
2842
Kron
S. J.
,
Spudich
J. A.
(
1986
).
Fluorescent actin filaments move on myosin fixed to a glass surface.
Proc. Nat. Acad. Sci. USA
83
,
6272
6276
Kron
S. J.
,
Toyoshima
Y. Y.
,
Uyeda
T. Q. P.
,
Spudich
J. A.
(
1991
).
Assays for actin sliding movement over myosin-coated surfaces.
Meth. Enzymol
196
,
399
416
Mimori
Y.
,
Miki-Noumura
T.
(
1994
).
ATP-induced sliding of microtubules on tracks of 22S dynein molecules aligned with the same polarity.
Cell Motil. Cytoskel
27
,
180
191
Moss
A. G.
,
Gatti
J.-L.
,
Witman
G. B.
(
1992
).
The motile/IC1 subunit of sea urchin sperm outer dynein does not form a rigor bond.
J. Cell Biol
118
,
1177
1188
Moss
A. G.
,
Sale
W. S.
,
Fox
L. A.
,
Witman
G. B.
(
1992
).
Thesubunit of sea urchin sperm outer arm dynein mediates structural and rigor binding to microtubules.
J. Cell Biol
118
,
1189
1200
Paschal
B. M.
,
King
S. M.
,
Moss
A. G.
,
Collins
C. A.
,
Vallee
R. B.
,
Witman
G. B.
(
1987
).
Isolated flagellar outer arm dynein translocates brain microtubules in vitro.
Nature
330
,
672
674
Reedy
M. C.
,
Beall
C.
,
Fyrberg
E.
(
1989
).
Formation of reverse rigor chevrons by myosin heads.
Nature
339
,
481
483
Sale
W. S.
,
Satir
P.
(
1977
).
Direction of active sliding of microtubules in Tetrahymena cilia.
Proc. Nat. Acad. Sci. USA
74
,
2045
2049
Sale
W. S.
,
Gibbons
I. R.
(
1979
).
Study of the mechanism of vanadate inhibition of the dynein cross-bridge cycle in sea urchin sperm flagella.
J. Cell Biol
82
,
291
298
Sale
W. S.
,
Fox
L. A.
(
1988
).
Isolated-heavy chain subunit of dynein translocates microtubules in vitro.
J. Cell Biol
107
,
1793
1797
Sellers
J. R.
,
Kachar
B.
(
1990
).
Polarity and velocity of sliding filaments: Control of direction by actin and of speed by myosin.
Science
249
,
406
408
Shimizu
T.
,
Johnson
K. A.
(
1983
).
Presteady state kinetic analysis of vanadate-induced inhibition of the dynein ATPase.
J. Biol. Chem
258
,
13833
13840
Shingyoji
C.
,
Murakami
A.
,
Takahashi
K.
(
1977
).
Local reactivation of Triton-extracted flagella by iontophoretic application of ATP.
Nature
265
,
269
270
Spudich
J. A.
,
Kron
S. J.
,
Sheetz
M. P.
(
1985
).
Movement of myosin-coated beads on oriented filaments reconstituted from purified actin.
Nature
315
,
584
586
Summers
K. E.
,
Gibbons
I. R.
(
1971
).
Adenosine triphosphate-induced sliding of tubules in trypsin-treated flagella of sea-urchin sperm.
Proc. Nat. Acad. Sci. USA
68
,
3092
3096
Vale
R. D.
,
Toyoshima
Y. Y.
(
1988
).
Rotation and translocation of microtubules in vitro induced by dyneins from Tetrahymena cilia.
Cell
52
,
459
469
Vallee
R. B.
(
1986
).
Reversible assembly purification of microtubules without assembly-promoting agents and further purification of tubulin, microtubule-associated proteins, and MAP fragments.
Meth. Enzymol
134
,
89
104
Warner
F. D.
,
Mitchell
D. R.
(
1981
).
Polarity of dynein-microtubule interactions in vitro: Cross-bridging between parallel and antiparallel microtubules.
J. Cell Biol
89
,
35
44
Yamada
A.
,
Ishii
N.
,
Takahashi
K.
(
1990
).
Direction and speed of actin filaments moving along thick filaments isolated from molluscan smooth muscle.
J. Biochem
108
,
341
343
Yamada
A.
,
Wakabayashi
T.
(
1993
).
Movement of actin away from the center of reconstituted rabbit myosin filament is slower than in the opposite direction.
Biophys. J
64
,
565
569
Yamada
A.
,
Yoshio
M.
,
Nakayama
H.
(
1997
).
Bi-directional movement of actin filaments along long bipolar tracks of oriented rabbit skeletal muscle myosin molecules.
FEBS Lett
409
,
380
384
Yano
Y.
,
Miki-Noumura
T.
(
1980
).
Sliding velocity between outer doublet microtubules of sea-urchin sperm axonemes.
J. Cell Sci
44
,
169
186
This content is only available via PDF.