Role of oviduct glycans in sperm binding and fertilization

Volz, Larissa J.

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https://hdl.handle.net/2142/115664 Copy

Description

Title

Role of oviduct glycans in sperm binding and fertilization

Author(s)

Volz, Larissa J.

Issue Date

2022-03-03

Director of Research (if dissertation) or Advisor (if thesis)

Miller, David J

Committee Member(s)

• Knox, Robert
• Dean, Matthew

Department of Study

Animal Sciences

Discipline

Animal Sciences

Degree Granting Institution

University of Illinois at Urbana-Champaign

Degree Name

M.S.

Degree Level

Thesis

Keyword(s)

• Reproduction
• oviduct
• in vitro fertilization

Abstract

The sperm reservoir in the oviduct plays an important role in sperm binding and extending sperm lifespan between semen deposition and ovulation, particularly when ovulation is delayed after semen deposition. 3-O-sulfated Lewis X trisaccharide (suLeX) and a biantennary glycan with 6-sialylated N-acetyllactosamine (bi-SiaLN) at its termini are found in the porcine oviduct and bind sperm with high specificity. Both glycans, when immobilized on beads, lengthen the lifespan of bound sperm compared to free-swimming sperm. However, it can be challenging to separate sperm bound to beads from free sperm. By coupling biotinylated glycans to streptavidin coverslips or streptavidin slides, we immobilized glycans to a two-dimensional surface to use this system to determine the effect of binding to immobilized suLeX on sperm fertilizing ability. Sperm were allowed to bind immobilized glycans to examine glycan-binding specificity and the effects of sperm concentration and binding time on the number of sperm bound to the glycan-coated coverslips and slides. Compared to the medium control, significantly more sperm bound to suLeX and the positive control, immobilized fibronectin. No binding was observed to the control glycan, N-acetyl-D-lactosamine (LacNAc). The number of sperm bound to immobilized suLeX increased proportionally with the concentration of sperm incubated. Sperm bound to immobilized glycans maximally within 30 minutes. Slides or coverslips gave similar results. This system allowed us to replicate the oviduct environment and test if binding to suLeX could extend sperm lifespan, so they would still fertilize oocytes after longer incubation times and improve IVF success. Sperm were incubated with immobilized suLeX, suLeA, fibronection, or medium for 0, 6, 12, or 24 hours. Pre-incubating with immobilized suLeX reduced the number of polyspermic zygotes from 41.3% in the control to 17.7%. After incubating sperm for 24 hours, only 0.01% of oocytes were fertilized in the control while 12 % of oocytes were fertilized with sperm pre-bound to immobilized suLeX. These results suggest that IVF success could be increased by using sperm pre-bound to immobilized suLeX. These systems allow more detailed study of the effects of sperm binding to oviduct glycans before fertilization.

Graduation Semester

2022-05

Type of Resource

Thesis

Copyright and License Information

Copyright 2022 Larissa Volz

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