The major sperm protein (MSP)-based amoeboid motility of Ascaris suum sperm requires coordinated lamellipodial protrusion and cell body retraction. In these cells, protrusion and retraction are tightly coupled to the assembly and disassembly of the cytoskeleton at opposite ends of the lamellipodium. Although polymerization along the leading edge appears to drive protrusion, the behavior of sperm tethered to the substrate showed that an additional force is required to pull the cell body forward. To examine the mechanism of cell body movement, we used pH to uncouple cytoskeletal polymerization and depolymerization. In sperm treated with pH 6.75 buffer, protrusion of the leading edge slowed dramatically while both cytoskeletal disassembly at the base of the lamellipodium and cell body retraction continued. At pH 6.35, the cytoskeleton pulled away from the leading edge and receded through the lamellipodium as its disassembly at the cell body continued. The cytoskeleton disassembled rapidly and completely in cells treated at pH 5.5, but reformed when the cells were washed with physiological buffer. Cytoskeletal reassembly occurred at the lamellipodial margin and caused membrane protrusion, but the cell body did not move until the cytoskeleton was rebuilt and depolymerization resumed. These results indicate that cell body retraction is mediated by tension in the cytoskeleton, correlated with MSP depolymerization at the base of the lamellipodium.
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6 September 1999
Article|
September 06 1999
Localized Depolymerization of the Major Sperm Protein Cytoskeleton Correlates with the Forward Movement of the Cell Body in the Amoeboid Movement of Nematode Sperm
Joseph E. Italiano, Jr.,
Joseph E. Italiano, Jr.
aDepartment of Biological Science, Florida State University, Tallahassee, Florida 32306
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Murray Stewart,
Murray Stewart
bMedical Research Council Laboratory of Molecular Biology, Cambridge CB2 2QH, United Kingdom
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Thomas M. Roberts
Thomas M. Roberts
aDepartment of Biological Science, Florida State University, Tallahassee, Florida 32306
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Joseph E. Italiano, Jr.
aDepartment of Biological Science, Florida State University, Tallahassee, Florida 32306
Murray Stewart
bMedical Research Council Laboratory of Molecular Biology, Cambridge CB2 2QH, United Kingdom
Thomas M. Roberts
aDepartment of Biological Science, Florida State University, Tallahassee, Florida 32306
1.used in this paper: HKB-acetate, HKB buffer containing 20 mM sodium acetate; MSP, major sperm protein; PAO, phenylarsine oxide
Dr. Italiano's current address is Division of Hematology, Brigham and Women's Hospital, Harvard Medical School, 221 Longwood Avenue, Boston, MA 02115.
Received:
February 10 1999
Revision Requested:
July 30 1999
Accepted:
August 03 1999
Online ISSN: 1540-8140
Print ISSN: 0021-9525
© 1999 The Rockefeller University Press
1999
The Rockefeller University Press
J Cell Biol (1999) 146 (5): 1087–1096.
Article history
Received:
February 10 1999
Revision Requested:
July 30 1999
Accepted:
August 03 1999
Citation
Joseph E. Italiano, Murray Stewart, Thomas M. Roberts; Localized Depolymerization of the Major Sperm Protein Cytoskeleton Correlates with the Forward Movement of the Cell Body in the Amoeboid Movement of Nematode Sperm. J Cell Biol 6 September 1999; 146 (5): 1087–1096. doi: https://doi.org/10.1083/jcb.146.5.1087
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