Role of carbohydrates in the attachment of equine spermatozoa to uterine tubal (oviductal) epithelial cells in vitro.
Abstract: To test the hypotheses that the attachment of equine spermatozoa to uterine tubal (oviductal) epithelial cells (OEC) in vitro is mediated by glycoproteins, and that proteins with carbohydrate-binding properties are present in the periacrosomal plasma membrane of equine spermatozoa. Methods: 4 reproductively sound stallions, and 1 mare in estrus. Methods: In experiment 1a, fluorescent-labeled spermatozoa were cocultured with monolayers of OEC in the presence of 50 mM glucose, fructose, galactose, mannose, N-acetyl glucosamine, N-acetyl galactosamine, or N-acetyl neuraminic acid, or 10 mg of fetuin or asialofetuin/ml in modified Tyrode's solution (TALP), or in TALP alone. After 2 hours of coculture, numbers of attached spermatozoa were counted by fluorescence microscopy and analysis of digitized images. In experiment 1b, progressive motility, viability, acrosomal integrity, and capacitation status were determined in spermatozoa incubated for 2 hours in the presence of the respective monosaccharides and glycoproteins or in TALP alone. In experiment 2, proteins isolated from the periacrosomal plasma membrane of equine spermatozoa were subjected to galactose affinity chromatography and subsequent one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis and silver staining. Results: Numbers of spermatozoa attached to OEC were reduced (P < 0.05) after all treatments except N-acetyl glucosamine, compared with incubation in TALP alone. The lowest numbers of spermatozoa were bound in cultures incubated in the presence of galactose and asialofetuin. Spermatozoal motility was lower (P < 0.05) after incubation for 2 hours in the presence of fetuin, compared with control, and incubation in the presence of fetuin or asialofetuin caused a significant (P < 0.05) increase in the percentage of capacitated spermatozoa, compared with control. Affinity chromatography of periacrosomal plasma membrane proteins revealed a galactose-binding protein of about 66 kd. Conclusions: Recognition of glycoconjugates with exposed galactosyl residues on OEC by galactose-binding proteins on the periacrosomal plasma membrane of equine spermatozoa could mediate the attachment of equine spermatozoa to OEC in vitro.
Publication Date: 1996-11-01 PubMed ID: 8915444
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research article investigates the role of carbohydrates in the attachment of horse sperm to uterine tubal cells. The study suggests that proteins with carbohydrate-binding properties on the sperm help mediate this connection.
Research Methods: Experiments and Observations
- The study was carried out in two primary experiments. In the first (1a), labeled sperm were grown with a layer of OEC (oviductal epithelium cells) in various solutions including glucose, fructose, galactose, mannose, N-acetyl glucosamine, N-acetyl galactosamine, N-acetyl neuraminic acid, fetuin and asialofetuin.
- After 2 hours of co-culturing, the numbers of attached sperm were counted and analyzed using fluorescence microscopy.
- In the second part of experiment 1 (1b), the researchers examined motility, viability, integrity, and capacitation status of the sperm after being incubated for 2 hours in the presence of various monosaccharides and glycoproteins.
- In the final experiment (2), the periacrosomal plasma membrane proteins of horse sperm were isolated and examined through galactose affinity chromatography and gel electrophoresis methods for further analysis.
Results of the Research
- The results showed that the number of sperm attached to OEC was reduced after all treatments except for N-acetyl glucosamine as compared to the control group.
- The lowest number of sperm attachments occurred in the presence of galactose and asialofetuin.
- In terms of sperm motility, the researchers noted a decrease in sperm movement after 2 hours of incubation in the presence of fetuin.
- There was also an increase in the percentage of capacitated (primed for fertilization) sperm after incubation in the presence of fetuin or asialofetuin.
- The research further revealed a galactose-binding protein of about 66 kd in size present in the sperm’s periacrosomal plasma membrane.
Conclusion of the Study
- The study concludes that glycoconjugates, a type of molecule made up of a carbohydrate and a protein, with exposed galactosyl residues on the OEC may be recognized by galactose-binding proteins on the sperm.
- This interaction may mediate the attachment of horse sperm to the OEC, providing a possible explanation for the role of carbohydrates in the attachment process.
Cite This Article
APA
Dobrinski I, Ignotz GG, Thomas PG, Ball BA.
(1996).
Role of carbohydrates in the attachment of equine spermatozoa to uterine tubal (oviductal) epithelial cells in vitro.
Am J Vet Res, 57(11), 1635-1639.
Publication
Researcher Affiliations
- Department of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, USA.
MeSH Terms
- Acrosome / physiology
- Animals
- Carbohydrates / physiology
- Cell Membrane / physiology
- Epithelium
- Fallopian Tubes / physiology
- Female
- Glycoproteins / physiology
- Horses / physiology
- In Vitro Techniques
- Male
- Monosaccharides / metabolism
- Sperm Motility / physiology
- Spermatozoa / physiology
Citations
This article has been cited 10 times.- Camara Pirez M, Li S, Koelle S. Assessment of Sperm Binding Capacity in the Tubal Reservoir Using a Bovine Ex Vivo Oviduct Culture and Fluorescence Microscopy. Methods Protoc 2021 Sep 23;4(4).
- Gimeno BF, Bariani MV, Laiz-Quiroga L, Martínez-León E, Von-Meyeren M, Rey O, Mutto AÁ, Osycka-Salut CE. Effects of In Vitro Interactions of Oviduct Epithelial Cells with Frozen-Thawed Stallion Spermatozoa on Their Motility, Viability and Capacitation Status. Animals (Basel) 2021 Jan 3;11(1).
- Camara Pirez M, Steele H, Reese S, Kölle S. Bovine sperm-oviduct interactions are characterized by specific sperm behaviour, ultrastructure and tubal reactions which are impacted by sex sorting. Sci Rep 2020 Oct 5;10(1):16522.
- Winters RA, Hamilton DN, Bhatnagar AS, Fitzgerald R, Bovin N, Miller DJ. Porcine sperm binding to oviduct cells and glycans as supplements to traditional laboratory semen analysis. J Anim Sci 2018 Dec 3;96(12):5265-5275.
- Miller DJ. Review: The epic journey of sperm through the female reproductive tract. Animal 2018 Jun;12(s1):s110-s120.
- Suarez SS. Mammalian sperm interactions with the female reproductive tract. Cell Tissue Res 2016 Jan;363(1):185-194.
- Tecle E, Gagneux P. Sugar-coated sperm: Unraveling the functions of the mammalian sperm glycocalyx. Mol Reprod Dev 2015 Sep;82(9):635-50.
- Coy P, García-Vázquez FA, Visconti PE, Avilés M. Roles of the oviduct in mammalian fertilization. Reproduction 2012 Dec;144(6):649-60.
- Henkel R. Sperm preparation: state-of-the-art--physiological aspects and application of advanced sperm preparation methods. Asian J Androl 2012 Mar;14(2):260-9.
- Miller DJ. Sperm in the Mammalian Female Reproductive Tract: Surfing Through the Tract to Try to Beat the Odds. Annu Rev Anim Biosci 2024 Feb 15;12:301-319.
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