Mouse Oviduct Specific Glycoprotein (OGP) is an Egg-Associated ZP3-Independent Sperm Adhesion Ligand Open Access

Lyng, Robert J (2009)

Permanent URL: https://etd.library.emory.edu/concern/etds/t722h959x?locale=en
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Abstract


Abstract
Mouse Oviduct Specific Glycoprotein (OGP) is an Egg-Associated ZP3-Independent
Sperm Adhesion Ligand

By Robert John Lyng
Mouse sperm-egg binding requires a multiplicity of receptor-ligand interactions,
including an oviduct-derived, basic, high molecular weight, wheat germ agglutinin
(WGA)-binding, glycoprotein that associates with the egg coat after ovulation. Herein we
report the purification and identification of a sperm-binding ligand. WGA-reactive, high
molecular weight glycoproteins isolated from superovulated mouse oviduct lysates
competitively inhibit sperm-egg binding in vitro. Within this heterogeneous glycoprotein
preparation, a distinct 220 kDa protein selectively binds to sperm surfaces. Sequencing of
the 220 kDa protein identified it as Oviduct-Specific Glycoprotein (OGP). OGP was
confirmed as a sperm-binding ligand by the following three results: specific immuno-
depletion of OGP from the denatured oviduct lysates completely abolished the sperm-
binding activity, the immunoprecipitated OGP was capable of competitively inhibiting
sperm-egg binding, and natively purified OGP also competitively inhibits sperm-egg
binding. As expected, the secretory cells of the oviduct fimbriae and infundibulum
express OGP; however, contrary to previous reports, OGP is found associated with both
the zona pellucida and the perivitelline space of oocytes. Western blot analysis and lectin
affinity chromatography demonstrates that whereas the bulk of OGP remains soluble in
the ampullar fluid, distinct OGP glycoforms associate with the cumulus matrix, zona
pellucida and perivitelline space. In this regard, the OGP sperm-binding activity is
restricted to a PNA-reactive glycoform that shows preferential binding to the sperm
surface, zona pellucida and perivitelline space, relative to other more abundant
glycoforms that do not localize to these domains nor have sperm-binding activity.
Interestingly, pretreatment of 2-cell embryos, which do not normally bind sperm, with
PNA-reactive OGP stimulates sperm binding. Finally, the sperm-binding activity of the
PNA-reactive OGP is shown to be carbohydrate dependent.

Table of Contents







Table of Contents

CHAPTER 1: INTRODUCTION
1
1.1 Introduction to Dissertation
1
1.2 Mammalian
Fertilization
2
1.2.1 Overview
2
1.2.2 Sperm-Oocyte Interactions
7
1.2.2.1 Sperm-ZP recognition and binding
7
1.2.2.1.1 Zona Pellucida 3 (ZP3)
7
1.2.2.1.2 1,4-Galactosyltransferase 1 (GalT 1)
8
1.2.2.1.3 SED1
10
1.2.2.1.4 Cyritestin
11
1.2.2.1.5 Glycodelins and Fucosyltransferase 5 (FUT5)
12
1.2.2.1.6 Arylsulfatase-A and SGG glycolipid
13
1.2.2.2 Sperm-ZP secondary interactions
14
1.2.2.2.1 Zona Pellucida 2 (ZP2)
14
1.2.2.2.2 sp56
15
1.2.2.2.3 Zonadhesin
16
1.2.2.3 Sperm-egg plasma membrane binding and fusion
17
1.2.2.3.1 CD9
17
1.2.2.3.2 Izumo
18
1.2.2.3.3 CRISP-1
19
1.2.2.3.4 SLLP-1
20

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