The Stiffness of the Flagella of Impaled Bull Sperm

Lindemann, Charles B.; Rudd, Walter G.; Rikmenspoel, Robert

doi:10.1016/s0006-3495(73)85997-1

Cited by 57 publications

(69 citation statements)

References 9 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…mation and propagation occur within an organelle that is otherwise relatively inactive and pliant. It is also supported by micromanipulation studies (10,39,46) and by our hydrodynamic shear experiments, which indicate that beating cilia are readily dissociable compared with rigor cilia.…”

Section: Relaxedsupporting

confidence: 69%

Substructure of the outer dynein arm

Goodenough¹

1982

The Journal of Cell Biology

191

120

View full text Add to dashboard Cite

The substructure of the outer dynein arm has been analyzed in quick-frozen deepetch replicas of Tetrahymena and Chlamydornonas axonemes. Each arm is found to be composed of five morphologically discrete components: an elliptical head; two spherical feet; a slender stalk; and an interdynein linker. The feet make contact with the A microtubule of each doublet; the stalk contacts the B microtubule; the head lies between the feet and stalk; and the linker associates each arm with its neighbor. The spatial relationships between these five components are found to be distinctly different in rigor (ATP-depleted) versus relaxed (ATP-or vanadate plus ATP-treated) axonemes, and the stalk appears to alter its affinity for the B microtubule in the relaxed state. Images of living cilia attached to Tetrahymena cells show that the relaxed configuration is adopted in vivo. We relate our observations to morphological and experimental studies reported by others and propose several models that suggest how this newly described dynein morphology may relate to dynein function. Doublet microtubules of eukaryotic cilia and flagella are bridged by outer dynein arms that are presumed to play a key role in the mechanochemical generation of ciliary motility (see references 6, 16, and 54 for recent reviews). Considerable progress has been made in analyzing the polypeptide components and the ATPase activities associated with the outer arms (11,49,64), and both biochemical and morphological studies indicate that each arm must be composed of several subunits (19,49,64,70). To date, however, the number, size, and spatial relationships of these postulated subunits remain unknown, largely because the arms are poorly resolved when viewed by traditional thin section or negative-stain electron microscopy. Two factors are likely to contribute to this poor resolution: the arms may well be sensitive to such manipulations as fLxation, dehydration, and air drying; moreover, because they repeat along each doublet with a 24-nm period and span a 21-nm inter-doublet gap, there is likely to be considerable image overlap in both thin-sectioned and negatively stained preparations.The quick-freeze, deep-etch technique (24) is well suited to the study of structures such as dynein arms, since material can be examined without chemical fLxation, dehydration, or staining, and the resultant three-dimensional platinum replicas are free of image superimposition. When care is taken to examine only material that is optimally frozen, meaningful images of high resolution can be obtained.Using this technique, we have analyzed the substructure of outer dynein arms in axonemes (demembranated cilia) of Tetrahymena and Chlamydomonas. We show that each outer arm consists of five discrete structural components, and we document the distinct configurations adopted by these subunits 798 under two extreme conditions: rigor, in which the axonemes are fully depleted of ATP, and relaxation, in which the axonemes are presented with ATP plus sodium vanadate, a dynein ATPase inhibit...

show abstract

Section: Relaxedsupporting

confidence: 69%

Substructure of the outer dynein arm

Goodenough¹

1982

The Journal of Cell Biology

191

120

View full text Add to dashboard Cite

show abstract

“…Relaxation of rigor bends, and low stiffness values, are only observed in the presence of MgATP2-. Some earlier reports (13,29,35) suggested that the relaxed state could be obtained with ADP; however, in my experiments, no relaxation was obtained with ADP if the adenylate kinase activity was inhibited and ATP contamination was excluded by diadenosinepenta-716 THE JOURNAL OF CELL BIOLOGY " VOLUME 85, 1980 phosphate and by hexokinase, respectively . The possibility that diadenosinepentaphosphate could inhibit the relaxation was examined by using the working solutions containing RBASE, 2.8 mM MgC1 2, 1.8 mM EGTA, 0.5 mM CaC1 2, 100 AM diadenosinepentaphosphate, and either 3 jM ATP or 2 mM unpurified ADP (which may contain several percent of ATP; purchased from Boehringer Mannheim Biochemicals).…”

Section: Rigor Bend Relaxationmentioning

confidence: 53%

“…(29) . Direct stiffness measurements on echinoderm sperm flagella (38) indicated that the relaxed state could be obtained with live spermatozoa immobilized by CO :, and that the rigor state could be obtained with demembranated spermatozoa in the absence of ATP.…”

Section: Rigor and Relaxed States Of Flagellamentioning

confidence: 99%

See 1 more Smart Citation

Inhibition and relaxation of sea urchin sperm flagella by vanadate.

Okuno¹

1980

The Journal of Cell Biology

View full text Add to dashboard Cite

Direct measurements of the stiffness (elastic bending resistance) of demembranated sea urchin sperm flagella were made in the presence of MgATP 2-and vanadate. Under these conditions, the flagellum is in a relaxed state, with a stiffness of -0 .9 x 10-2 ' N m l, which is -5% of the stiffness obtained in the rigor state in the absence of MgATP2-. MgADP -'does not substitute for MgATP 2 " in producing the relaxed state . A progressive inhibition of movement is observed after addition of MgATP 2-to flagella preincubated with vanadate, in which new bend generation, propagation, and relaxation by straightening are distinguished, depending on the ratio of MgATP 2-and vanadate. At appropriate concentrations of vanadate, increase of the velocity of bend propagation is observed at a very low concentration of MgATP 2-that is not enough to induce spontaneous beating . Vanadate enhances competitive inhibition of beat frequency by MgADP -but not by ADP3-, ATP -, or Pi. These observations, and the uncompetitive inhibition of beat frequency by vanadate, indicate that vanadate can only bind to dyneinnucleotide complexes induced by MgATP 2-and MgADP -. The state accessible by MgATP2-binding must be a state in which the cross-bridges are detached and the flagellum is relaxed . The state accessible by MgADP -binding must be a crossbridged state . Bound vanadate prevents the transition between these two states . Inhibition and relaxation by vanadate in the presence of MgATP2-results from the specific affinity of vanadate for a state in which nucleotide is bound, rather than a specific affinity for the detached state .The sliding microtubule hypothesis for flagellar and ciliary movement has been firmly established by a series of experimental (42, 49, 52) and theoretical works (4,5,31, 40), since the first important observations by P. Satir (43,44) . This hypothesis predicts that the dynein arm plays a vital role in generating the bending wave of flagella, because the dynein arm is involved in the transduction of the chemical energy provided by ATP dephosphorylation into mechanical energy (15, 16) . The conformational change of the dynein arm is assumed to involve the cyclic change of its orientation, including association and dissociation with the adjacent microtubule, as in the actomyosin system (26). This mechanochemical behavior of the dynein arm is thought to be tightly coupled with the cyclic dephosphorylation of ATP, because the rate of ATP hydrolysis is tightly coupled with flagellar movement and beat frequency (7, 10, 11), and because dynein ATPase not only has a latency (I, 21, 32, 47, 48) but also can be increased in activity by microtubular protein (l, 39) and tubulin (25).The dynein microtubule coupling mediated with ATP has also been suggested by some physiologi-J . CELL . BIOLOGY

show abstract

“…Mechanical studies of flagellar stiffness seemed to support this view. When flagella are in the relaxed condition, which is induced by either vanadate inhibition of the dynein (Lindemann et al 2005, Pelle et al 2009) or inhibition of dynein by 10 mM ATP (Lindemann et al 1973), the flagella are in their most flexible state and their stiffness is minimized.…”

Section: Seeing Is Believingmentioning

confidence: 99%

The geometric clutch at 20: stripping gears or gaining traction?

Lindemann

Lesich

2015

Reproduction

View full text Add to dashboard Cite

It has been 20 years since the geometric clutch (GC) hypothesis was first proposed. The core principle of the GC mechanism is fairly simple. When the axoneme of a eukaryotic flagellum is bent, mechanical stress generates forces transverse to the outer doublets (t-forces). These t-forces can push doublets closer together or pry them apart. The GC hypothesis asserts that changes in the inter-doublet spacing caused by t-forces are responsible for the activation and deactivation of the dynein motors, that creates the beat cycle. A series of computer models utilizing the clutch mechanism has shown that it can simulate ciliary and flagellar beating. The objective of the present review is to assess where things stand with the GC hypothesis in the clarifying light of new information. There is considerable new evidence to support the hypothesis. However, it is also clear that it is necessary to modify some of the original conceptions of the hypothesis so that it can be consistent with the results of recent experimental and ultrastructural studies. In particular, dynein deactivation by t-forces must be able to occur with dyneins that remain attached to the B-subtubule of the adjacent doublet.Reproduction (2015) 150 R45-R53

show abstract

The Stiffness of the Flagella of Impaled Bull Sperm

Cited by 57 publications

References 9 publications

Substructure of the outer dynein arm

Substructure of the outer dynein arm

Inhibition and relaxation of sea urchin sperm flagella by vanadate.

The geometric clutch at 20: stripping gears or gaining traction?

Contact Info

Product

Resources

About