| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
Articles |
a Department of Biomedical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, New York 14853
b Department of Basic Sciences, Division of Biochemistry and
c Department of Herd Health and Reproduction, Faculty of Veterinary Medicine, Utrecht University, 3584 CL Utrecht, The Netherlands
In cattle, sperm are stored in a reservoir in the caudal isthmus of the oviduct until the time of ovulation approaches. Bull sperm are trapped in the reservoir by binding to fucosylated molecules on the oviductal epithelium. Capacitated sperm lose binding affinity for the epithelium; therefore this study was undertaken to determine whether this occurs because capacitated bull sperm lose binding affinity for fucose. BSA conjugated to 
-L-fucopyranosylphenyl isothiocyanate and fluorescein isothiocyanate (fuc-BSA-FITC) was used in conjunction with flow cytometry to monitor the capacity of bull sperm to bind fucose. Dead sperm were identified using ethidium homodimer and were excluded from analysis. BSA-FITC conjugated with mannose (man-BSA-FITC) and BSA-FITC were used as controls. When examined by epifluorescence microscopy, motile bull sperm that exhibited labeling by any of the probes were fluorescent over the acrosomal region of the plasma membrane. By flow cytometry, labeling of live sperm was greatest for sperm that had been washed in TALP medium and probed with fuc-BSA-FITC (mean ± SD:167 ± 6.0 relative fluorescence units, collected in logarithmic mode). Labeling by fuc-BSA-FITC was lower in unwashed sperm (60 ± 2.7) and in washed sperm with seminal plasma added back (56 ± 8.0). Labeling was also reduced by centrifuging washed sperm through a Percoll step gradient (103 ± 6.3) and by capacitating washed sperm in medium containing 10 µg/ml heparin (50 ± 4.4). BSA-FITC labeling was barely detectable in all treatments. Man-BSA-FITC produced little labeling of washed sperm (22 ± 0.6), as expected; however, intense labeling appeared over the acrosomal region of sperm incubated under capacitating conditions (128 ± 21.6). It was concluded that removal of seminal plasma exposes fucose-binding sites, which are then lost or modified during capacitation, thereby allowing the release of sperm from the reservoir. At that time, mannose-binding sites are revealed or activated, which might serve to bind sperm to the zona pellucida.
1 This project was supported by USDA NRICGP project number 32738 to S.S.S. and by the School of Veterinary Medicine, Utrecht University, The Netherlands.
2 Correspondence: S.S. Suárez, Department of Biomedical Sciences, T5-006 Veterinary Research Tower, Cornell University, Ithaca, NY 14853. FAX: 607 253 3541; sss7{at}cornell.edu
This article has been cited by other articles:
|
|
 |
|
  T. L. Tollner, A. I. Yudin, A. F. Tarantal, C. A. Treece, J. W. Overstreet, and G. N. Cherr Beta-Defensin 126 on the Surface of Macaque Sperm Mediates Attachment of Sperm to Oviductal Epithelia Biol Reprod, March 1, 2008; 78(3): 400 - 412. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. M. Gwathmey, G. G. Ignotz, J. L. Mueller, P. Manjunath, and S. S. Suarez Bovine Seminal Plasma Proteins PDC-109, BSP-A3, and BSP-30-kDa Share Functional Roles in Storing Sperm in the Oviduct Biol Reprod, October 1, 2006; 75(4): 501 - 507. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S.S. Suarez and A. A. Pacey Sperm transport in the female reproductive tract Hum. Reprod. Update, January 1, 2006; 12(1): 23 - 37. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Ekhlasi-Hundrieser, K. Gohr, A. Wagner, M. Tsolova, A. Petrunkina, and E. Topfer-Petersen Spermadhesin AQN1 Is a Candidate Receptor Molecule Involved in the Formation of the Oviductal Sperm Reservoir in the Pig Biol Reprod, September 1, 2005; 73(3): 536 - 545. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Sostaric, C. H.A. van de Lest, B. Colenbrander, and B. M. Gadella Dynamics of Carbohydrate Affinities at the Cell Surface of Capacitating Bovine Sperm Cells Biol Reprod, February 1, 2005; 72(2): 346 - 357. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. M. Gwathmey, G. G. Ignotz, and S. S. Suarez PDC-109 (BSP-A1/A2) Promotes Bull Sperm Binding to Oviductal Epithelium In Vitro and May Be Involved in Forming the Oviductal Sperm Reservoir Biol Reprod, September 1, 2003; 69(3): 809 - 815. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. Reeve, W.L. Ledger, and A.A. Pacey Does the Arg-Gly-Asp (RGD) adhesion sequence play a role in mediating sperm interaction with the human endosalpinx? Hum. Reprod., July 1, 2003; 18(7): 1461 - 1468. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Boilard, J. Bailey, S. Collin, M. Dufour, and M.-A. Sirard Effect of Bovine Oviduct Epithelial Cell Apical Plasma Membranes on Sperm Function Assessed by a Novel Flow Cytometric Approach Biol Reprod, October 1, 2002; 67(4): 1125 - 1132. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Khunsook, J. A. Alhadeff, and B. S. Bean Purification and characterization of human seminal plasma {alpha}-L-fucosidase Mol. Hum. Reprod., March 1, 2002; 8(3): 221 - 227. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
