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Use of microbeads for the detection of binding sites on the human zona pellucida: a scanning electron microscopy (SEM) assay.

Abstract：

:One prerequisite for fertilization is the specific binding of spermatozoa to the zona pellucida. However, the factors and mechanisms involved in this gamete contact are not well understood. Gamete recognition and binding are species-specific and are controlled by oligosaccharides of the zona and their corresponding carbohydrates on the spermatozoon. By using a specific lectin we developed a technique to detect those oligosaccharides on the human zona pellucida that might be involved in the binding process. Microbeads (Ø = 2.8 microm), used as artificial spermatozoa, were coated with lectin Con A and cultured together with 75 unfertilized oocytes (group A) remaining after intracytoplasmic sperm injection. Con A binds specifically to alpha-d-mannose and alpha-d-glucose. As a control, 75 unfertilized oocytes after intracytoplasmic sperm injection (group B) were also cultured together with Con A-covered microbeads, but in a medium containing a binding inhibiting sugar (alpha-methyl-mannopyrasosid). The number and distribution of the microbeads on human oocytes of both groups were analysed on scanning electron microscopy images. Beads on oocytes of group A had binding patterns similar to those of spermatozoa. They were distributed in an extremely heterogeneous way with various numbers of bound beads both on individual and different oocytes. Most of the group A oocytes (85%) had more than 50 beads bound to the zona, in contrast to the control oocytes of group B, where 68% had less than 10 bound beads. The use of an inhibiting sugar abolished the binding capacity of the microbeads nearly completely. This technique is a powerful tool for the detection of binding sites on the zona pellucida, i.e. those sugars that are responsible for contact between spermatozoa and the zona pellucida.

journal_name

Andrologia

journal_title

Andrologia

authors

Michelmann HW,Töpfer-Petersen E,Schwartz P,Gratz G,Magerkurth C

doi

10.1046/j.1439-0272.2001.00412.x

keywords:

subject

Has Abstract

pub_date

2001-09-01 00:00:00

pages

266-71

issue

eissn

0303-4569

issn

1439-0272

pii

412

journal_volume

pub_type

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Use of microbeads for the detection of binding sites on the human zona pellucida: a scanning electron microscopy (SEM) assay.