PMID: 9883571Jan 12, 1999Paper

Temperature-dependent flagellar motility of demembranated, cytosol-free fowl spermatozoa

Comparative Biochemistry and Physiology. Part A, Molecular & Integrative Physiology
K AshizawaY Tsuzuki

Abstract

A rapid and gentle procedure for preparing demembranated, cytosol-free sperm models was applied to fowl spermatozoa. Intact spermatozoa were introduced to a Triton X-100-containing extraction medium layered on top of a discontinuous Percoll gradient in a 1.5 ml microfuge tube. After brief exposure to the extraction medium, spermatozoa were separated from the plasma membrane and detergent-soluble components by centrifugation through a 55% Percoll layer, finally collecting on top of a 90% Percoll cushion from where they were recovered. Optimum conditions consisted of a Triton X-100 concentration in the extraction medium of 0.15%, duration of demembranation time of 1.5 min and ATP concentration in the reactivation medium of 0.5 mM. Demembranated sperm models obtained by this procedure could be reactivated, and the motility at 30 degrees C was more than 60%, but negligible at 40 degrees C. These values were similar to those obtained from the conventional method, in which centrifugation is not carried out, and which results in some of the cytosolic components being transferred to the reactivation medium along with the spermatozoa. Inhibition of motility was observed following the addition of EGTA or myosin light chain kinase (MLCK) ...Continue Reading

References

Jun 15, 1992·Biochemical and Biophysical Research Communications·K AshizawaY Tsuzuki
Aug 1, 1992·Journal of Reproduction and Fertility·K Ashizawa, G J Wishart
Nov 1, 1991·Journal of Reproduction and Fertility·M F Thomson, G J Wishart
May 1, 1989·Journal of Reproduction and Fertility·K AshizawaK Okauchi
Jan 1, 1989·Cell Motility and the Cytoskeleton·J S Tash
May 1, 1989·Journal of Reproduction and Fertility·K AshizawaK Okauchi
Jul 1, 1987·Journal of Reproduction and Fertility·G J Wishart, K Ashizawa
Nov 1, 1987·Journal of Cellular Biochemistry·C J Brokaw
Jan 1, 1994·Cell Motility and the Cytoskeleton·J T San Agustin, G B Witman
Oct 1, 1993·Molecular Reproduction and Development·H Mohri
Jan 1, 1993·Cell Motility and the Cytoskeleton·J T San Agustin, G B Witman

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