Characterization of the functions and proteomes associated with membrane rafts in chicken sperm

PloS One
Ai UshiyamaAtsushi Asano

Abstract

Cellular membranes are heterogeneous, and this has a great impact on cellular function. Despite the central role of membrane functions in multiple cellular processes in sperm, their molecular mechanisms are poorly understood. Membrane rafts are specific membrane domains enriched in cholesterol, ganglioside GM1, and functional proteins, and they are involved in the regulation of a variety of cellular functions. Studies of the functional characterization of membrane rafts in mammalian sperm have demonstrated roles in sperm-egg binding and the acrosomal reaction. Recently, our biochemical and cell biological studies showed that membrane rafts are present and might play functional roles in chicken sperm. In this study, we isolated membrane rafts from chicken sperm as a detergent-resistant membranes (DRM) floating on a density gradient in the presence of 1% Triton X-100, and characterized the function and proteomes associated with these domains. Biochemical comparison of the DRM between fresh and cryopreserved sperm demonstrated that cryopreservation induces cholesterol loss specifically from membrane rafts, indicating the functional connection with reduced post-thaw fertility in chicken sperm. Furthermore, using an avidin-biotin sy...Continue Reading

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Methods Mentioned

BETA
Assay
protein assay
density gradient centrifugation

Software Mentioned

Xcalibur
Sequest HT
PANTHER
Leica AF6000
SOSUI
Orbitrap Fusion Tune Application
LAS AF
ImageJ
Proteome Discoverer

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