Title

Sonication of mouse sperm membranes reveals distinct protein domains

Authors

Authors

S. S. Baker; R. A. Cardullo;C. D. Thaler

Comments

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Abbreviated Journal Title

Biol. Reprod.

Keywords

acrosome reaction; fertilization; gamete biology; signal transduction; sperm; ZONA-PELLUCIDA; EXTRACELLULAR-MATRIX; PLASMA-MEMBRANE; MAMMALIAN; FERTILIZATION; BOAR SPERMATOZOA; IN-VITRO; BINDING; CAPACITATION; HYALURONIDASE; ACTIVATION; Reproductive Biology

Abstract

Molecular interactions between sperm and zona pellucida (ZP) during mammalian fertilization are not well characterized. To begin to characterize sperm components that are involved in sperm-ZP interactions, we isolated and density fractionated sperm membranes. The membrane fractions recovered from a density fractionation protocol were characterized, and sonication was compared with vortexing for preparation of sperm membranes by examining the distribution of proteins in the membrane fractions obtained from these 2 protocols. Biochemical and microscopic analyses were used to determine the composition of the sonicated membrane fractions, and immunoblotting was used to identify fractions containing some of the previously suggested ZP3 receptors. Transmission electron microscopy revealed that bands 1-3 contained membrane vesicles and band 4 contained axonemal and midpiece fragments. SDS-PAGE revealed that bands 1 and 2 shared many proteins, but band 3 contained a number of unique proteins. Surface labeling with I-125 demonstrated that bands 1 and 2 contained the majority of the sperm surface protein markers, whereas band 3 contained minor amounts of surface markers. Lectin-binding characteristics of sperm membrane glycoproteins were used to compare the relative distribution of glycosylated proteins in vortexed or sonicated membrane preparations. These characterizations indicate that sonication enhanced the differential distribution of sperm membrane proteins among the density fractions and suggests that this method is preferable for preparation of membrane fractions to be used for identification of proteins that mediate sperm-egg interactions.

Journal Title

Biology of Reproduction

Volume

66

Issue/Number

1

Publication Date

1-1-2002

Document Type

Article

Language

English

First Page

57

Last Page

64

WOS Identifier

WOS:000172912800009

ISSN

0006-3363

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