We have utilized immunoblotting and light microscopic immunofluorescent staining methods to examine the expression and localization of sea urchin kinesin-II, a heterotrimeric plus end-directed microtubule motor protein (previously referred to as KRP(85/95)), in sea urchin and sand dollar sperm. We d
Mechanical properties of the passive sea urchin sperm flagellum
β Scribed by Pelle, Dominic W. ;Brokaw, Charles J. ;Lesich, Kathleen A. ;Lindemann, Charles B.
- Publisher
- John Wiley and Sons
- Year
- 2009
- Tongue
- English
- Weight
- 355 KB
- Volume
- 66
- Category
- Article
- ISSN
- 0886-1544
- DOI
- 10.1002/cm.20401
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β¦ Synopsis
Abstract
In this study we used Triton Xβ100 extracted sea urchin spermatozoa to investigate the mechanical behavior of the basic 9+2 axoneme. The dynein motors were disabled by vanadate so that the flagellum is rendered a passive structure. We find that when a proximal portion of the flagellum is bent with a glass microprobe, the remainder of the flagellum distal to the probe exhibits a bend in the opposite direction (a counterbend). The counterbend can be understood from the prevailing sliding doublet model of axoneme mechanics, but does require the existence of elastic linkages between the outer doublets. Analysis of the shapes of counterbends provides a consensus value of 0.03β0.08/ΞΌm^2^ for the ratio of the interdoublet shear resistance (E~S~) to the bending resistance (E~B~) and we find that the ratio E~S~/E~B~ is relatively conserved for both passive flagella and transiently quiescent live flagella. This ratio expresses a fundamental mechanical property of the eukaryotic axoneme. It defines the contributions to total bending resistance derived from bending the microtubules and from stretching the interdoublet linkages, respectively. Using this ratio, and computer simulations of earlier experiments that measured the total stiffness of the flagellum, we obtain estimates of approximately 1 Γ 10^8^ pN nm^2^/rad for E~B~ and 6 pN/rad for E~S~, assuming that both elasticities are linear. Our results indicate that the behavior of the flagellum is close to that predicted by a linear model for shear elasticity. Cell Motil. Cytoskeleton 2009. Β© 2009 WileyβLiss, Inc.
π SIMILAR VOLUMES
The movement parameters of a sea urchin sperm flagellum can be manipulated mechanically by applying various modes of periodic vibrations to the sperm head held by suction in the tip of a micropipette. The beat frequency of the flagellum readily synchronizes with the frequency of the externally impos